A Polyphenol-rich Pomegranate Fruit Extract Suppresses NF-kappaB and IL-6 Expression by Blocking the Activation of IKKbeta and NIK in Primary Human Chondrocytes.
chondrocytes; Chondrocytes – Drug Effects; Chondrocytes/drug effects; Fruit; Fruit/chemistry; Gene Expression Regulation/drug effects; Genes – Drug Effects; Humans; I-kappa B Kinase/genetics/*metabolism; IkappaB; IKKbeta; IL-1beta; Interleukin 1 – Metabolism; Interleukin-1beta/metabolism; Interleukin-6/genetics/*metabolism; Interleukins; Interleukins – Metabolism; NF-kappa B; NF-kappa B – Metabolism; NF-kappa B/genetics/*metabolism; NF-kappaB; NIK; osteoarthritis; Phosphorylation – Drug Effects; Phosphorylation/drug effects; Plant Extracts; Plant Extracts – Pharmacodynamics; Plant Extracts/chemistry/*pharmacology; Polyphenols – Pharmacodynamics; Polyphenols/pharmacology; pomegranate; Pomegranate; Protein-Serine-Threonine Kinases/genetics/*metabolism; Proteins – Metabolism; Punicaceae/*chemistry; Signal Transduction – Drug Effects; Signal Transduction/drug effects; Transcription Factor RelA/metabolism; Transferases; Transferases – Metabolism
Pomegranate fruit extract (PE) rich in polyphenols has been shown to exert chondroprotective effects, but the mechanism is not established. Here, we used an in vitro model of inflammation in osteoarthritis (OA) to investigate the potential of PE to suppress interleukin 1 beta (IL-1beta)-stimulated expression of inflammatory cytokine IL-6, generation of reactive oxygen species (ROS) levels, and investigated the mechanism of NF-kappaB inhibition by analyzing the activation of the kinases upstream of IkappaBalpha in primary human chondrocytes. Total and phosphorylated forms of kinases and expression of IL-6 were determined at protein and mRNA levels by western immunoblotting and Taqman assay, respectively. Dihydrorhodamine 123 staining estimated ROS generation. Pomegranate fruit extract inhibited the mRNA and protein expression of IL-6, generation of ROS, and inhibited the IL-1beta-mediated phosphorylation of inhibitor of nuclear factor kappa-B kinase subunit beta (IKKbeta), expression of IKKbeta mRNA, degradation of IkappaBalpha, and activation and nuclear translocation of
Haseeb Abdul; Khan Nazir M; Ashruf Omer S; Haqqi Tariq M
Phytotherapy research : PTR
2017
2017-05
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1002/ptr.5799" target="_blank" rel="noreferrer noopener">10.1002/ptr.5799</a>
A standardized extract of Butea monosperma (Lam.) flowers suppresses the
*Butea; Aged; Autophagy; Autophagy/drug effects/physiology; Butea monosperma (Lam.); Chondrocytes/drug effects/*metabolism; Dose-Response Relationship; Drug; Flowers; Gene Expression; Humans; Interleukin-1beta/*pharmacology; Interleukin-6/*biosynthesis/genetics; Matrix Metalloproteinase 13/biosynthesis/genetics; Matrix Metalloproteinase 3/biosynthesis/genetics; Matrix Metalloproteinase 9/biosynthesis/genetics; Matrix Metalloproteinases/*biosynthesis/genetics; Middle Aged; mTOR; Nutraceuticals; Osteoarthritis; Osteoarthritis/*metabolism; Plant Extracts/isolation & purification/pharmacology
BACKGROUND/OBJECTIVE: Osteoarthritis (OA) is a leading cause of joint dysfunction, disability and poor quality of life in the affected population. The underlying mechanism of joint dysfunction involves increased oxidative stress, inflammation, high levels of cartilage extracellular matrix degrading proteases and decline in autophagy-a mechanism of cellular defense. There is no disease modifying therapies currently available for OA. Different parts of the Butea monosperma (Lam.) plant have widely been used in the traditional Indian Ayurvedic medicine system for the treatment of various human diseases including inflammatory conditions. Here we studied the chondroprotective effect of hydromethanolic extract of Butea monosperma (Lam.) flowers (BME) standardized to the concentration of Butein on human OA chondrocytes stimulated with IL-1beta. METHODS: The hydromethanolic extract of Butea monosperma (Lam.) (BME) was prepared with 70% methanol-water mixer using Soxhlet. Chondrocytes viability after BME treatment was measured by MTT assay. Gene expression levels were determined by quantitative polymerase chain reaction (qPCR) using TaqMan assays and immunoblotting with specific antibodies. Autophagy activation was determined by measuring the levels of microtubule associated protein 1 light chain 3-II (LC3-II) by immunoblotting and visualization of autophagosomes by transmission electron and confocal microscopy. RESULTS: BME was non-toxic to the OA chondrocytes at the doses employed and suppressed the IL-1beta induced expression of inerleukin-6 (IL-6) and matrix metalloprotease-3 (MMP-3), MMP-9 and MMP-13. BME enhanced autophagy in chondrocytes as determined by measuring the levels of
Ansari Mohammad Y; Khan Nazir M; Haqqi Tariq M
Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie
2017
2017-12
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1016/j.biopha.2017.09.140" target="_blank" rel="noreferrer noopener">10.1016/j.biopha.2017.09.140</a>
A Wogonin-rich Fraction of scutellaria baicalensis Root Extract Exerts Chondroprotective Effects by Suppressing il-1β-induced Activation of AP-1 in Human OA Chondrocytes.
Khan N M; Haseeb A; Ansari M Y; Haqqi T M
Osteoarthritis & Cartilage
2017
2017-04-02
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1016/j.joca.2017.02.493" target="_blank" rel="noreferrer noopener">10.1016/j.joca.2017.02.493</a>
A wogonin-rich-fraction of Scutellaria baicalensis root extract exerts chondroprotective effects by suppressing IL-1beta-induced activation of AP-1 in human OA chondrocytes.
Cells; Chemical Fractionation/methods; Chondrocytes/*drug effects/metabolism; Cultured; Cyclooxygenase 2/genetics/metabolism; Flavanones/*pharmacology; Gene Expression Regulation/drug effects; Humans; Interleukin-1beta/pharmacology; Interleukin-6/genetics/metabolism; Osteoarthritis/pathology; Plant Extracts/*pharmacology; Plant Roots/*chemistry; Protective Agents/pharmacology; Reactive Oxygen Species/metabolism; Scutellaria baicalensis/*chemistry; Transcription Factor AP-1/genetics/*metabolism
Osteoarthritis (OA) is a common joint disorder with varying degrees of inflammation and sustained oxidative stress. The root extract of Scutellaria baicalensis (SBE) has been used for the treatment of inflammatory and other diseases. Here, we performed activity-guided HPLC-fractionation of SBE, identified the active ingredient(s) and investigated its chondroprotective potential. We found that the Wogonin containing fraction-4 (F4) was the most potent fraction based on its ability to inhibit ROS production and the suppression of catabolic markers including IL-6, COX-2, iNOS, MMP-3, MMP-9,
Khan Nazir M; Haseeb Abdul; Ansari Mohammad Y; Haqqi Tariq M
Scientific reports
2017
2017-03
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1038/srep43789" target="_blank" rel="noreferrer noopener">10.1038/srep43789</a>
An effective and efficient method of transfecting primary human chondrocytes in suspension.
*Chondrocytes; *Osteoarthritis; *siRNA; *Transfection; Aggrecans/genetics; Articular/metabolism/pathology; Cartilage; Cells; Chondrocytes/cytology/*metabolism; Collagen Type II/genetics; Cultured; Humans; Messenger/*genetics; mRNA Cleavage and Polyadenylation Factors/antagonists & inhibitors/genetics; Nucleotidyltransferases/antagonists & inhibitors/genetics; Plasmids/*genetics; RNA; Small Interfering/*genetics; Transfection/*methods
Human chondrocytes accumulate an ECM-rich matrix by secreting matrix macromolecules during monolayer culture, which makes them difficult to transfect efficiently. Here we report a non-viral based protocol to transfect the primary human chondrocytes with high efficiency in suspension. Chondrocyte cultures were digested using Pronase and Collagenase and transfected in suspension. Transfection efficiencies of more than 80% were achieved routinely using the protocol described. The viability of siRNA transfected or un-transfected chondrocytes was not affected and resulted in 80-90% knockdown of the target mRNA levels. This protocol may be useful in gene knockdown, and ectopic overexpression studies in chondrocytes.
Makki Mohammad Shahidul; Akhtar Nahid; Haqqi Tariq M
Analytical biochemistry
2017
2017-06
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1016/j.ab.2017.03.009" target="_blank" rel="noreferrer noopener">10.1016/j.ab.2017.03.009</a>
An operant-based detection method for inferring tinnitus in mice.
*Conditioning; *Disease Models; *Inferior colliculus; *Mouse model; *Noise-induced hearing loss; *Operant conditioning; *Sodium salicylate; *Tinnitus; Acoustic Stimulation; Analysis of Variance; Animal; Animals; Auditory; Avoidance Learning; Brain Stem/physiology; Electroshock; Equipment Design; Evoked Potentials; Female; Inbred C57BL; Inferior Colliculi/physiopathology; Male; Mice; Motor Activity; Neurons/physiology; Operant; Otoacoustic Emissions; Sodium Salicylate; Spontaneous/physiology; Tinnitus/*diagnosis/physiopathology; Tissue Culture Techniques; Voltage-Sensitive Dye Imaging
BACKGROUND: Subjective tinnitus is a hearing disorder in which a person perceives sound when no external sound is present. It can be acute or chronic. Because our current understanding of its pathology is incomplete, no effective cures have yet been established. Mouse models are useful for studying the pathophysiology of tinnitus as well as for developing therapeutic treatments. NEW METHOD: We have developed a new method for determining acute and chronic tinnitus in mice, called sound-based avoidance detection (SBAD). The SBAD method utilizes one paradigm to detect tinnitus and another paradigm to monitor possible confounding factors, such as motor impairment, loss of motivation, and deficits in learning and memory. RESULTS: The SBAD method has succeeded in monitoring both acute and chronic tinnitus in mice. Its detection ability is further validated by functional studies demonstrating an abnormal increase in neuronal activity in the inferior colliculus of mice that had previously been identified as having tinnitus by the SBAD method. COMPARISON WITH EXISTING METHODS: The SBAD method provides a new means by which investigators can detect tinnitus in a single mouse accurately and with more control over potential confounding factors than existing methods. CONCLUSION: This work establishes a new behavioral method for detecting tinnitus in mice. The detection outcome is consistent with functional validation. One key advantage of mouse models is they provide researchers the opportunity to utilize an extensive array of genetic tools. This new method could lead to a deeper understanding of the molecular pathways underlying tinnitus pathology.
Zuo Hongyan; Lei Debin; Sivaramakrishnan Shobhana; Howie Benjamin; Mulvany Jessica; Bao Jianxin
Journal of neuroscience methods
2017
2017-11
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1016/j.jneumeth.2017.08.029" target="_blank" rel="noreferrer noopener">10.1016/j.jneumeth.2017.08.029</a>
Animal Models for Dysphagia Studies: What Have We Learnt So Far.
*Animal models; *Deglutition; *Deglutition disorders; *Disease Models; *Pathophysiology; *Performance; Animal; Animals; Biological; Biomedical Research/*methods; Deglutition – Physiology; Deglutition Disorders – Physiopathology; Deglutition Disorders/*physiopathology; Deglutition/physiology; Humans; Medical – Methods; Models; Research
Research using animal models has contributed significantly to realizing the goal of understanding dysfunction and improving the care of patients who suffer from dysphagia. But why should other researchers and the clinicians who see patients day in and day out care about this work? Results from studies of animal models have the potential to change and grow how we think about dysphagia research and practice in general, well beyond applying specific results to human studies. Animal research provides two key contributions to our understanding of dysphagia. The first is a more complete characterization of the physiology of both normal and pathological swallow than is possible in human subjects. The second is suggesting of specific, physiological, targets for development and testing of treatment interventions to improve dysphagia outcomes.
German Rebecca Z; Crompton A W; Gould Francois D H; Thexton Allan J
Dysphagia
2017
2017-02
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1007/s00455-016-9778-7" target="_blank" rel="noreferrer noopener">10.1007/s00455-016-9778-7</a>
Beyond thermoregulation: metabolic function of cetacean blubber in migrating bowhead and beluga whales.
*Lipid Metabolism; Adipose Tissue/*metabolism; Aging/metabolism; Amino Acid Sequence; Animals; Base Sequence; Beluga Whale/*physiology; Blubber; Body Temperature Regulation; Bowhead whale; Bowhead Whale/*physiology; Development; Female; Humans; Inbred C57BL; Leptin; Leptin/genetics; Leptin/genetics/metabolism; Lipase/genetics; Long-Evans; Male; Metabolic activity; Mice; Rats; Receptors; Seasons
The processes of lipid deposition and utilization, via the gene leptin (Lep), are poorly understood in taxa with varying degrees of adipose storage. This study examines how these systems may have adapted in marine aquatic environments inhabited by cetaceans. Bowhead (Balaena mysticetus) and beluga whales (Delphinapterus leucas) are ideal study animals-they possess large subcutaneous adipose stores (blubber) and undergo bi-annual migrations concurrent with variations in food availability. To answer long-standing questions regarding how (or if) energy and lipid utilization adapted to aquatic stressors, we quantified variations in gene transcripts critical to lipid metabolism related to season, age, and blubber depth. We predicted leptin tertiary structure conservation and assessed inter-specific variations in Lep transcript numbers between bowheads and other mammals. Our study is the first to identify seasonal and age-related variations in Lep and lipolysis in these cetaceans. While Lep transcripts and protein oscillate with season in adult bowheads reminiscent of hibernating mammals, transcript levels reach up to 10 times higher in bowheads than any other mammal. Data from immature bowheads are consistent with the hypothesis that short baleen inhibits efficient feeding. Lipolysis transcripts also indicate young Fall bowheads and those sampled during Spring months limit energy utilization. These novel data from rarely examined species expand the existing knowledge and offer unique insight into how the regulation of Lep and lipolysis has adapted to permit seasonal deposition and maintain vital blubber stores.
Ball H C; Londraville R L; Prokop J W; George John C; Suydam R S; Vinyard C; Thewissen J G M; Duff R J
Journal of comparative physiology. B, Biochemical, systemic, and environmental physiology
2017
2017-01
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1007/s00360-016-1029-6" target="_blank" rel="noreferrer noopener">10.1007/s00360-016-1029-6</a>
Brief Stimulus Exposure Fully Remediates Temporal Processing Deficits Induced by Early Hearing Loss.
Female; Male; Animals; Age Factors; *auditory cortex; *development; *gap detection; *hearing loss; *remediation; *temporal coding; Acoustic Stimulation/*methods; Auditory Cortex/*physiology/*physiopathology; Auditory Perception/*physiology; Gerbillinae; Hearing Loss/*physiopathology; Brain Stem/*physiology; Evoked Potentials; Auditory
In childhood, partial hearing loss can produce prolonged deficits in speech perception and temporal processing. However, early therapeutic interventions targeting temporal processing may improve later speech-related outcomes. Gap detection is a measure of auditory temporal resolution that relies on the auditory cortex (ACx), and early auditory deprivation alters intrinsic and synaptic properties in the ACx. Thus, early deprivation should induce deficits in gap detection, which should be reflected in ACx gap sensitivity. We tested whether earplugging-induced, early transient auditory deprivation in male and female Mongolian gerbils caused correlated deficits in behavioral and cortical gap detection, and whether these could be rescued by a novel therapeutic approach: brief exposure to gaps in background noise. Two weeks after earplug removal, animals that had been earplugged from hearing onset throughout auditory critical periods displayed impaired behavioral gap detection thresholds (GDTs), but this deficit was fully reversed by three 1 h sessions of exposure to gaps in noise. In parallel, after earplugging, cortical GDTs increased because fewer cells were sensitive to short gaps, and gap exposure normalized this pattern. Furthermore, in deprived animals, both first-spike latency and first-spike latency jitter increased, while spontaneous and evoked firing rates decreased, suggesting that deprivation causes a wider range of perceptual problems than measured here. These cortical changes all returned to control levels after gap exposure. Thus, brief stimulus exposure, perhaps in a salient context such as the unfamiliar placement into a testing apparatus, rescued impaired gap detection and may have potential as a remediation tool for general auditory processing deficits.SIGNIFICANCE STATEMENT Hearing loss in early childhood leads to impairments in auditory perception and language processing that can last well beyond the restoration of hearing sensitivity. Perceptual deficits can be improved by training, or by acoustic enrichment in animal models, but both approaches involve extended time and effort. Here, we used a novel remediation technique, brief periods of auditory stimulus exposure, to fully remediate cortical and perceptual deficits in gap detection induced by early transient hearing loss. This technique also improved multiple cortical response properties. Rescue by this efficient exposure regime may have potential as a therapeutic tool to remediate general auditory processing deficits in children with perceptual challenges arising from early hearing loss.
Green David B; Mattingly Michelle M; Ye Yi; Gay Jennifer D; Rosen Merri J
The Journal of neuroscience : the official journal of the Society for Neuroscience
2017
2017-08
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1523/JNEUROSCI.0916-17.2017" target="_blank" rel="noreferrer noopener">10.1523/JNEUROSCI.0916-17.2017</a>
Dataset of effect of Wogonin, a natural flavonoid, on the viability and activation of NF-kappaB and MAPKs in IL-1beta-stimulated human OA chondrocytes.
MAPK; Mass-spectrometry; NF-kappaB; Nrf2; Osteoarthritis; Wogonin
This article contains data related to the article "Wogonin, a plant derived small molecule exerts potent anti-inflammatory and chondroprotective effects through activation of ROS/ERK/Nrf2 signaling pathways in human Osteoarthritis chondrocytes" (Khan et al. 2017) [1]. The data are related to effects of Wogonin on the viability and IL-1beta-stimulated activation of NF-kappaB and ERK1/2, JNK1/2 and p38 MAPKs in human OA chondrocytes. Gene expression data representing the chondrogenic phenotype and the efficiency of Nrf2 knockdown in monolayer culture of human OA chondrocytes were shown. Moreover, mass spectrometric calibration curve of Wogonin used to quantify the intracellular uptake were also presented. The data are presented in the form of figures and significance of these has been given in the research article (Khan et al. 2017) [1].
Khan Nazir M; Haseeb Abdul; Ansari Mohammad Y; Devarapalli Pratap; Haynie Sara; Haqqi Tariq M
Data in brief
2017
2017-06
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1016/j.dib.2017.03.054" target="_blank" rel="noreferrer noopener">10.1016/j.dib.2017.03.054</a>
Deep sequencing and analyses of miRNAs, isomiRs and miRNA induced silencing complex (miRISC)-associated miRNome in primary human chondrocytes.
MicroRNAs, a group of small, noncoding RNAs that post-transcriptionally regulate gene expression, play important roles in chondrocyte function and in the development of osteoarthritis. We characterized the dynamic repertoire of the chondrocyte miRNome and miRISC-associated miRNome by deep sequencing analysis of primary human chondrocytes. IL-1beta treatment showed a modest effect on the expression profile of miRNAs in normal and osteoarthritis (OA) chondrocytes. We found a number of miRNAs that showed a wide range of sequence modifications including nucleotide additions and deletions at 5' and 3' ends; and nucleotide substitutions. miR-27b-3p showed the highest expression and miR-140-3p showed the highest number of sequence variations. AGO2 RIP-Seq analysis revealed the differential recruitment of a subset of expressed miRNAs and isoforms of miRNAs (isomiRs) to the miRISC in response to IL-1beta, including miR-146a-5p, miR-155-5p and miR-27b-3p. Together, these results reveal a complex repertoire of miRNAs and isomiRs in primary human chondrocytes. Here, we also show the changes in miRNA composition of the miRISC in primary human chondrocytes in response to
Haseeb Abdul; Makki Mohammad Shahidul; Khan Nazir M; Ahmad Imran; Haqqi Tariq M
Scientific reports
2017
2017-11
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1038/s41598-017-15388-4" target="_blank" rel="noreferrer noopener">10.1038/s41598-017-15388-4</a>
Elimination of dysfunctional mitochondria by parkin suppresses oxidative stress and expression of osteoarthritis related genes in human chondrocytes.
Ansari M Y; Khan N M; Ahmad I; Haqqi T M
Osteoarthritis & Cartilage
2017
2017-04-02
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1016/j.joca.2017.02.086" target="_blank" rel="noreferrer noopener">10.1016/j.joca.2017.02.086</a>
Evolutionary aspects of the development of teeth and baleen in the bowhead whale.
*Biological Evolution; Animals; baleen; baleen whales; bowhead whale; Bowhead Whale/anatomy & histology/*embryology; Cetacea; Dentition; embryology; Female; FGF; Jaw/anatomy & histology/embryology; keratin; Mixed; Mouth/anatomy & histology/*embryology; mysticetes; ontogeny; Pregnancy; tooth development; Tooth/anatomy & histology/*embryology
In utero, baleen whales initiate the development of several dozens of teeth in upper and lower jaws. These tooth germs reach the bell stage and are sometimes mineralized, but toward the end of prenatal life they are resorbed and no trace remains after birth. Around the time that the germs disappear, the keratinous baleen plates start to form in the upper jaw, and these form the food-collecting mechanism. Baleen whale ancestors had two generations of teeth and never developed baleen, and the prenatal teeth of modern fetuses are usually interpreted as an evolutionary leftover. We investigated the development of teeth and baleen in bowhead whale fetuses using histological and immunohistochemical evidence. We found that upper and lower dentition initially follow similar developmental pathways. As development proceeds, upper and lower tooth germs diverge developmentally. Lower tooth germs differ along the length of the jaw, reminiscent of a heterodont dentition of cetacean ancestors, and lingual processes of the dental lamina represent initiation of tooth bud formation of replacement teeth. Upper tooth germs remain homodont and there is no evidence of a secondary dentition. After these germs disappear, the oral epithelium thickens to form the baleen plates, and the protein FGF-4 displays a signaling pattern reminiscent of baleen plates. In laboratory mammals, FGF-4 is not involved in the formation of hair or palatal rugae, but it is involved in tooth development. This leads us to propose that the signaling cascade that forms teeth in most mammals has been exapted to be involved in baleen plate ontogeny in mysticetes.
Thewissen J G M; Hieronymus Tobin L; George John C; Suydam Robert; Stimmelmayr Raphaela; McBurney Denise
Journal of anatomy
2017
2017-04
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1111/joa.12579" target="_blank" rel="noreferrer noopener">10.1111/joa.12579</a>
Harpagoside suppresses IL-6 expression in primary human osteoarthritis chondrocytes.
*c-FOS/AP-1; *harpagoside; *IL-6; *MMP-13; *osteoarthritis; CCAAT-Enhancer-Binding Protein-beta/metabolism; Chemokines/metabolism; Chondrocytes/*drug effects/metabolism; Drug Evaluation; Glycosides/pharmacology/*therapeutic use; Harpagophytum; Humans; Interleukin-1beta; Interleukin-6/antagonists & inhibitors/*metabolism; Matrix Metalloproteinase 13/metabolism; NF-kappa B/metabolism; Osteoarthritis/*drug therapy/metabolism; Phytotherapy; Plant Extracts/pharmacology/therapeutic use; Preclinical; Primary Cell Culture; Proto-Oncogene Proteins c-fos/metabolism; Pyrans/pharmacology/*therapeutic use; Reactive Oxygen Species/metabolism; Transcription Factor AP-1/metabolism
There is growing evidence in support of the involvement of inflammatory response in the pathogenesis of osteoarthritis (OA). Harpagoside, one of the bioactive components of Harpagophytum procumbens (Hp), has been shown to possess anti-inflammatory properties. Here we used an in vitro model of inflammation in OA to investigate the potential of harpagoside to suppress the production of inflammatory cytokines/chemokines such as IL-6 and matrix degrading proteases. We further investigated the likely targets of harpagoside in primary human OA chondrocytes. OA chondrocytes were pre-treated with harpagoside before stimulation with IL-1beta. mRNA expression profile of 92 cytokines/chemokines was determined using TaqMan Human Chemokine PCR Array. Expression levels of selected mRNAs were confirmed using TaqMan assays. Protein levels of IL-6 and MMP-13 were assayed by ELISA and immunoblotting. Total protein levels and phosphorylation of signaling proteins were determined by immunoblotting. Cellular localization of
Haseeb Abdul; Ansari Mohammad Yunus; Haqqi Tariq M
Journal of orthopaedic research : official publication of the Orthopaedic Research Society
2017
2017-02
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1002/jor.23262" target="_blank" rel="noreferrer noopener">10.1002/jor.23262</a>
Histone deacetylase inhibitor vorinostat (SAHA, MK0683) perturb miR-9-MCPIP1 axis to block IL-1beta-induced IL-6 expression in human OA chondrocytes.
*IL-6; *MCPIP1; *miRNA; *Osteoarthritis; *Vorinostat; Adult; Aged; CCAAT-Enhancer-Binding Proteins/genetics/metabolism; Cells; Chondrocytes/*metabolism/pathology; Cultured; Female; Gene Expression Regulation/*drug effects; Genetic; Humans; Hydroxamic Acids/*pharmacology; Interleukin-1beta/genetics/*metabolism; Interleukin-6/genetics/*metabolism; Male; MicroRNAs/genetics/*metabolism; Middle Aged; Osteoarthritis/drug therapy/genetics/*metabolism/pathology; Promoter Regions; Ribonucleases/*biosynthesis/genetics; Transcription Factors/*biosynthesis/genetics; Vorinostat
AIM OF THE STUDY: High levels of IL-6 are believed to contribute to osteoarthritis (OA) pathogenesis. The expression of IL-6 is regulated post-transcriptionally by the miR-9-MCPIP-1 axis in chondrocytes. Vorinostat (SAHA) inhibits the IL-6 expression in OA chondrocytes. We investigated whether SAHA suppresses the expression of IL-6 by perturbing the miR-9-MCPIP1 axis in OA chondrocytes under pathological conditions. MATERIALS AND METHODS: OA chondrocytes were isolated by enzymatic digestion and treated with IL-1beta in the absence or presence of SAHA. Genes and protein expression levels were determined by TaqMan assays and Western blotting, respectively. Secreted IL-6 was quantified by enzyme linked immunosorbent assay (ELISA). MCPIP1 promoter deletion mutants were generated by polymerase chain reaction (PCR). Promoter recruitment of transcription factors was determined by ChIP. Nuclear run-on was employed to measure the ongoing transcription. siRNA-mediated knockdown of the CEBPalpha expression was employed for loss of function studies. RESULTS: Expression of MCPIP1 was high in SAHA treated OA chondrocytes but expression of IL-6 mRNAs and secreted IL-6 were reduced by \textasciitilde70%. SAHA suppressed the expression of miR-9 but enhanced the activity of the MCPIP1 promoter localized to a 156bp region which also harbors the binding site for CEBPalpha. Treatment with SAHA enhanced the recruitment of CEBPalpha to the MCPIP1 promoter. Ectopically expressed CEBPalpha enhanced the promoter activity and the expression of MCPIP1 while siRNA-mediated knockdown of CEBPalpha inhibited the expression of MCPIP1. CONCLUSIONS: Taken together our data indicate that SAHA-mediated suppression of the IL-6 expression is achieved through increased recruitment of CEBPalpha to the MCPIP1 promoter and by relieving the miR-9-mediated inhibition of MCPIP1 expression in OA chondrocytes.
Makki Mohammad S; Haqqi Tariq M
Connective tissue research
2017
2017-01
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1080/03008207.2016.1211113" target="_blank" rel="noreferrer noopener">10.1080/03008207.2016.1211113</a>
Identification of Novel Agents for the Treatment of Brain Metastases of Breast Cancer.
Female; Humans; Animals; Mice; Apoptosis; Cell Line; ADME; Antineoplastic Agents/*therapeutic use; brain cancer; Brain Neoplasms/*drug therapy/pathology/*secondary; Breast Neoplasms/*pathology; chemotherapy; CNS; distribution; drug discovery; Drug resistance; Tumor
BACKGROUND: Brain cancer from metastasized breast cancer has a high mortality rate in women. The treatment of lesions is hampered in large part by the blood-brain barrier (BBB), which prevents adequate distribution of anti-cancer compounds to brain metastases. METHOD: In this study we used a novel screening method to identify candidate molecules that are well-suited to utilizing the BBB choline transporter for distribution into the brain parenchyma. RESULTS: From our screen we identified two compounds, Ch-1 and Ch-2 that were able to reduce the brain tumor burden in a murine mouse model of brain metastasis of breast cancer. These compounds also significantly increased the survival of mice by more than 10 days. Mechanistic studies indicated that Ch-1 is able to prevent the activation of the pro-survival mitogen-activated kinases (MAPKs) by osteoactivin (OA; Glycoprotein nonmetastatic melanoma protein B GPNMB). CONCLUSION: The results from this study show that nutrient transporter virtual screening is a viable novel alternative to traditional drug screening programs to identify anti-cancer compounds for the treatment of brain cancers.
Venishetty Vinay K; Geldenhuys Werner J; Terell-Hall Tori B; Griffith Jessica I G; Sondag Gregory R; Safadi Fayez F; Lockman Paul R
Current cancer drug targets
2017
1905-7
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.2174/1568009617666161121123948" target="_blank" rel="noreferrer noopener">10.2174/1568009617666161121123948</a>
Inhibition of cartilage degradation and suppression of PGE2 and MMPs expression by pomegranate fruit extract in a model of posttraumatic osteoarthritis.
*Phytotherapy; *Punicaceae; ACLT; Animal; Animals; Anterior Cruciate Ligament/drug effects/metabolism/pathology; Apoptosis; Cartilage/cytology/*drug effects/metabolism/pathology; Chondrocytes/drug effects/metabolism/pathology; Collagen Type II/genetics/metabolism; Dinoprostone/*metabolism; Disease Models; Disease Progression; Female; Fruit; Interleukins/metabolism; Joints/cytology/*drug effects/metabolism/pathology; Male; Messenger/metabolism; Metalloproteases/genetics/*metabolism; Mitogen-Activated Protein Kinases/metabolism; MMPs; NF-kappa B/metabolism; Osteoarthritis; Osteoarthritis/*drug therapy/etiology/metabolism/pathology; PGE(2); Plant Extracts/pharmacology/therapeutic use; Pomegranate; Rabbit; Rabbits; RNA; Synovial Fluid/metabolism
OBJECTIVE: Osteoarthritis (OA) is characterized by cartilage degradation in the affected joints. Pomegranate fruit extract (PFE) inhibits cartilage degradation in vitro. The aim of this study was to determine whether oral consumption of PFE inhibits disease progression in rabbits with surgically induced OA. METHODS: OA was surgically induced in the tibiofemoral joints of adult New Zealand White rabbits. In one group, animals were fed PFE in water for 8 wk postsurgery. In the second group, animals were fed PFE for 2 wk before surgery and for 8 wk postsurgery. Histologic assessment and scoring of the cartilage was per Osteoarthritis Research Society International guidelines. Gene expression and matrix metalloproteinases (MMP) activity were determined using quantitative reverse transcriptase polymerase chain reaction and fluorometric assay, respectively. Interleukin (IL)-1 beta, MMP-13, IL-6, prostaglandin (PG)E2, and type II collagen (COL2A1) levels in synovial fluid/plasma/culture media were quantified using enzyme-linked immunosorbent assay. Expression of active caspase-3 and poly (ADP-ribose) polymerase p85 was determined by immunohistochemistry. Effect of PFE and inhibitors of MMP-13, mitogen-activated protein kinase (MAPK) and nuclear factor (NF)-kappaB was studied in IL-1 beta-stimulated rabbit articular chondrocytes. RESULTS: Safranin-O-staining and chondrocyte cluster formation was significantly reduced in the anterior cruciate ligament transaction plus PFE fed groups. Expression of MMP-3, MMP-9, and MMP-13 mRNA was higher in the cartilage of rabbits given water alone but was significantly lower in the animals fed PFE. PFE-fed rabbits had lower IL-6,
Akhtar Nahid; Khan Nazir M; Ashruf Omer S; Haqqi Tariq M
Nutrition (Burbank, Los Angeles County, Calif.)
2017
2017-01
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1016/j.nut.2016.08.004" target="_blank" rel="noreferrer noopener">10.1016/j.nut.2016.08.004</a>
Long-Lasting forward Suppression of Spontaneous Firing in Auditory Neurons: Implication to the Residual Inhibition of Tinnitus.
acoustic trauma; Animals; Brain Stem – Physiology; Clinical Assessment Tools; Cochlear Nerve – Physiology; Cochlear Nerve/*physiology; Inbred CBA; Inferior Colliculi/*physiology; inferior colliculus; Male; mice; Mice; residual inhibition; Sound; Tinnitus – Physiopathology; Tinnitus/*physiopathology
Tinnitus is the perception of a sound that has no external source. Sound stimuli can suppress spontaneous firing in auditory neurons long after stimulus offset. It is unknown how changes in sound stimulus parameters affect this forward suppression. Using in vivo extracellular recording in awake mice, we found that about 40 % of spontaneously active inferior colliculus (IC) neurons exhibited forward suppression of spontaneous activity after sound offset. The duration of this suppression increased with sound duration and lasted about 40 s following a
Galazyuk A V; Voytenko S V; Longenecker R J
Journal of the Association for Research in Otolaryngology : JARO
2017
2017-04
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1007/s10162-016-0601-9" target="_blank" rel="noreferrer noopener">10.1007/s10162-016-0601-9</a>
Metabotropic glutamate and GABA receptors modulate cellular excitability and glutamatergic transmission in chicken cochlear nucleus angularis neurons.
*Cellular excitability; *GABA(B)R; *mGluR; *Neuromodulation; *Nucleus angularis; *Synaptic transmission; Action Potentials/drug effects; Animals; Chick Embryo; Cochlear Nucleus/cytology/*physiology; Excitatory Postsynaptic Potentials/drug effects; GABA-B Receptor Agonists/pharmacology; GABA-B/*physiology; Glutamic Acid/*physiology; Metabotropic Glutamate/agonists/classification/*physiology; Neurons/drug effects/physiology; Patch-Clamp Techniques; Receptors; Synaptic Transmission/drug effects
Neurons in the avian cochlear nucleus angularis (NA) receive glutamatergic input from the auditory nerve, and GABAergic input from the superior olivary nucleus. Physiologically heterogeneous, NA neurons perform multiple functions including encoding sound intensity information. Using in vitro whole-cell patch recordings from acute brain slices and immunohistochemistry staining, we investigated neuromodulation mediated by metabotropic glutamate and GABA receptors (mGluRs and GABABRs) in NA neurons. Based on their intrinsic firing patterns in response to somatic current injections, NA neurons were classified into onset, damped, and tonic cells. Pharmacological activation of group II mGluRs, group III mGluRs, and GABABRs, by their respective agonists, suppressed the cellular excitability of non-onset firing NA neurons. Each of these agonists inhibited the glutamatergic transmission in NA neurons, in a cell type-independent manner. The frequency but not the amplitude of spontaneous release of glutamate was reduced by each of these agonists, suggesting that the modulation of the glutamatergic transmission was via presynaptic actions. Interestingly, activation of group I mGluRs increased cellular excitability and suppressed glutamatergic transmission in non-onset neurons. These results elaborate that auditory processing in NA neurons is subject to neuromodulation mediated by metabotropic receptors activated by native neurotransmitters released at NA.
Shi Wei; Lu Yong
Hearing research
2017
2017-03
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1016/j.heares.2017.01.011" target="_blank" rel="noreferrer noopener">10.1016/j.heares.2017.01.011</a>
Noise-induced cochlear synaptopathy: Past findings and future studies.
*Auditory Perception; *Hearing; *Hearing loss; *Molecular approach; *Preclinical model; *Spiral ganglion; *Synaptic loss; *Synaptic Transmission; Animals; Auditory; Hair Cells; Hearing Loss; Hearing Tests; Humans; Inner/*pathology; Noise-Induced/diagnosis/*pathology/physiopathology/psychology; Noise/*adverse effects; Predictive Value of Tests; Psychoacoustics; Spiral Ganglion/*pathology/physiopathology; Synapses/*pathology
For decades, we have presumed the death of hair cells and spiral ganglion neurons are the main cause of hearing loss and difficulties understanding speech in noise, but new findings suggest synapse loss may be the key contributor. Specifically, recent preclinical studies suggest that the synapses between inner hair cells and spiral ganglion neurons with low spontaneous rates and high thresholds are the most vulnerable subcellular structures, with respect to insults during aging and noise exposure. This cochlear synaptopathy can be "hidden" because this synaptic loss can occur without permanent hearing threshold shifts. This new discovery of synaptic loss opens doors to new research directions. Here, we review a number of recent studies and make suggestions in two critical future research directions. First, based on solid evidence of cochlear synaptopathy in animal models, it is time to apply molecular approaches to identify the underlying molecular mechanisms; improved understanding is necessary for developing rational, effective therapies against this cochlear synaptopathy. Second, in human studies, the data supporting cochlear synaptopathy are indirect although rapid progress has been made. To fully identify changes in function that are directly related this hidden synaptic damage, we argue that a battery of tests including both electrophysiological and behavior tests should be combined for diagnosis of "hidden hearing loss" in clinical studies. This new approach may provide a direct link between cochlear synaptopathy and perceptual difficulties.
Kobel Megan; Le Prell Colleen G; Liu Jennifer; Hawks John W; Bao Jianxin
Hearing research
2017
2017-06
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1016/j.heares.2016.12.008" target="_blank" rel="noreferrer noopener">10.1016/j.heares.2016.12.008</a>
Pre-pharyngeal Swallow Effects of Recurrent Laryngeal Nerve Lesion on Bolus Shape and Airway Protection in an Infant Pig Model.
*Aspiration; *Deglutition; *Deglutition disorders; *Infant; *Oro-pharyngeal pathophysiology; *Recurrent laryngeal nerve; Animal Studies; Animals; Aspiration – Etiology; Aspiration/etiology; Deglutition – Physiology; Deglutition Disorders – Etiology; Deglutition Disorders – Physiopathology; Deglutition Disorders/*etiology/physiopathology; Deglutition/physiology; Laryngeal Nerve Injuries/*complications; Laryngeal Nerves – Injuries; Newborn; Pneumonia; Swine
Recurrent laryngeal nerve (RLN) damage in infants leads to increased dysphagia and aspiration pneumonia. Recent work has shown that intraoral transport and swallow kinematics change following RLN lesion, suggesting potential changes in bolus formation prior to the swallow. In this study, we used geometric morphometrics to understand the effect of bolus shape on penetration and aspiration in infants with and without RLN lesion. We hypothesized (1) that geometric bolus properties are related to airway protection outcomes and (2) that in infants with RLN lesion, the relationship between geometric bolus properties and dysphagia is changed. In five infant pigs, dysphagia in 188 swallows was assessed using the Infant Mammalian Penetration-Aspiration Scale (IMPAS). Using images from high-speed VFSS, bolus shape, bolus area, and tongue outline were quantified digitally. Bolus shape was analyzed using elliptical Fourier analysis, and tongue outline using polynomial curve fitting. Despite large inter-individual differences, significant within individual effects of bolus shape and bolus area on airway protection exist. The relationship between penetration-aspiration score and both bolus area and shape changed post lesion. Tongue shape differed between pre- and post-lesion swallows, and between swallows with different IMPAS scores. Bolus shape and area affect airway protection outcomes. RLN lesion changes that relationship, indicating that proper bolus formation and control by the tongue require intact laryngeal sensation. The impact of RLN lesion on dysphagia is pervasive.
Gould Francois D H; Yglesias B; Ohlemacher J; German R Z
Dysphagia
2017
2017-06
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1007/s00455-016-9762-2" target="_blank" rel="noreferrer noopener">10.1007/s00455-016-9762-2</a>
Sucrose, But Not Glucose, Blocks IL1-beta-Induced Inflammatory Response in Human Chondrocytes by Inducing Autophagy via AKT/mTOR Pathway.
*AUTOPHAGY; *CHONDROCYTES; *OSTEOARTHRITIS; *SUCROSE; Adult; Aged; Autophagy/*drug effects; Chondrocytes/*metabolism/pathology; Female; Glucose/*pharmacology; Humans; Inflammation/chemically induced/metabolism/pathology; Interleukin-1beta/*pharmacology; Male; Middle Aged; Osteoarthritis/*metabolism/pathology; Proto-Oncogene Proteins c-akt/*metabolism; Signal Transduction/*drug effects; Sucrose/*pharmacology; TOR Serine-Threonine Kinases/*metabolism
Pathogenesis of osteoarthritis (OA) is multifactorial but interleukin-1beta (IL-1beta) is known to be an important mediator of cartilage degradation. Autophagy is an essential cellular homeostasis mechanism and has been proposed to protect against cartilage degradation and chondrocyte death under pathological conditions. We investigated the role of autophagy activated by sucrose, a natural disaccharide, in suppressing inflammatory mediator's expression and cell death under pathological conditions in human chondrocytes. Autophagy activation was investigated by Western blotting for LC3 and Beclin-1, immunofluorescence staining for LC3 puncta, and measuring autophagic flux. Activation of mTOR, AKT, and P70S6K was evaluated by Western blotting. Chondrocyte apoptosis was evaluated by propidium iodide (PI) staining using flowcytometry, expression of Bax by Western blotting, gene expression by TaqMan assays and caspase 3/7 activity was measured using a luminescence-based assay. We found that sucrose-induced active autophagy in OA chondrocytes in vitro was dependent on the activation of AKT/mTOR/P70S6K signaling pathways but was independent of reactive oxygen species (ROS) production. Sucrose activated autophagy blocked IL-1beta-induced apoptosis and mRNA expression of MMP-13, COX-2, and IL-6 in human OA chondrocytes. Glucose or fructose, the two metabolites of sucrose, failed to induce autophagy indicating that autophagy was specifically mediated by sucrose. In conclusion, sucrose attenuated IL-1beta induced apoptosis and the expression of catabolic mediators by inducing autophagy, and the autophagy in part was mediated through the activation of AKT/mTOR/P70S6K signaling pathway in human OA chondrocytes. J. Cell. Biochem. 118: 629-639, 2017. (c) 2016 Wiley Periodicals, Inc.
Khan Nazir M; Ansari Mohammad Y; Haqqi Tariq M
Journal of cellular biochemistry
2017
2017-03
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1002/jcb.25750" target="_blank" rel="noreferrer noopener">10.1002/jcb.25750</a>
Surprising absence of heparin in the intestinal mucosa of baby pigs.
*antibiotics; *heparin; *intestine; *microbiome; *pig; Animals; Anticoagulants/*metabolism; Chondroitin Sulfates/metabolism; Heparin/biosynthesis/isolation & purification/*metabolism; Heparitin Sulfate/metabolism; Intestinal Mucosa/*metabolism; Mast Cells/metabolism; Newborn/*metabolism; Swine
Heparin, a member of a family of molecules called glycosaminoglycans, is biosynthesized in mucosal mast cells. This important anticoagulant polysaccharide is primarily produced by extraction of the mast cell-rich intestinal mucosa of hogs. There is concern about our continued ability to supply sufficient heparin to support the worldwide growth of advanced medical procedures from the static population of adult hogs used as food animals. While the intestinal mucosa of adult pigs is rich in anticoagulant heparin (containing a few hundred milligrams per animal), little is known about how the content of heparin changes with animal age. Using sophisticated mass spectral analysis we discovered that heparin was largely absent from the intestinal mucosa of piglets. Moreover, while the related, nonanticoagulant heparan sulfate glycosaminoglycan was present in significant amounts we found little chondroitin sulfate E also associated with mast cells. Histological evaluation of piglet intestinal mucosa showed a very low mast cell content. Respiratory mast cells have been reported in baby pigs suggesting that there was something unique about the piglets used in the current study. These piglets were raised in the relatively clean environment of a university animal facility and treated with antibiotics over their lifetime resulting in a depleted microbiome that greatly reduced the number of mast cells and heparin content of the intestinal mucosal in these animals. Thus, from the current study it remains unclear whether the lack of intestinal mast cell-derived heparin results from the young age of these animals or their exposure to their depleted microbiome.
Yu Yanlei; Chen Yin; Mikael Paiyz; Zhang Fuming; Stalcup Apryll M; German Rebecca; Gould Francois; Ohlemacher Jocelyn; Zhang Hong; Linhardt Robert J
Glycobiology
2017
2017-01
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1093/glycob/cww104" target="_blank" rel="noreferrer noopener">10.1093/glycob/cww104</a>
Wogonin exerts anti-inflammatory effect by disrupting keap-1/Nrf2 interactions and activating Nrf2 in human OA chondrocytes.
Khan N M; Ansari M Y; Haynie S; Haqqi T M
Osteoarthritis & Cartilage
2017
2017-04-02
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1016/j.joca.2017.02.063" target="_blank" rel="noreferrer noopener">10.1016/j.joca.2017.02.063</a>
Wogonin exerts anti-inflammatory effect by disrupting KEAP-1/NRF2 interactions and activating NRF2 in human OA chondrocytes.
Khan N M; Ansari M Y; Haynie S; Haqqi T M
Osteoarthritis & Cartilage
2017
2017-04-02
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1016/j.joca.2017.02.775" target="_blank" rel="noreferrer noopener">10.1016/j.joca.2017.02.775</a>
Wogonin, a natural flavonoid, intercalates with genomic DNA and exhibits protective effects in IL-1beta stimulated osteoarthritis chondrocytes.
Apoptosis/drug effects; Binding Sites; Cells; Chondrocytes/cytology/*drug effects/metabolism; Chondroprotective effects; Cultured; Denaturation; DNA binding; DNA/chemistry/*metabolism; Flavanones/chemistry/metabolism/*pharmacology; Flavonoids/chemistry/pharmacology; Fluorescence Resonance Energy Transfer; Humans; Intercalating Agents/chemistry/metabolism/*pharmacology; Interleukin-1beta/*pharmacology; Molecular Docking Simulation; Nucleic Acid Conformation; Osteoarthritis; Osteoarthritis/metabolism/pathology; Protective Agents/chemistry/metabolism/*pharmacology; Reactive Oxygen Species/metabolism; Up-Regulation/drug effects; Wogonin
Wogonin has recently been shown to possess anti-inflammatory and chondroprotective properties and is of considerable interest due to its broad pharmacological activities. The present study highlights that Wogonin binds DNA and exerts chondroprotective effects in vitro. Wogonin showed strong binding with chondrocytes genomic DNA in vitro. The mode of binding of Wogonin to genomic-DNA was assessed by competing Wogonin with EtBr or DAPI, known DNA intercalator and a minor groove binder, respectively. EtBr fluorescence reduced significantly with increase in Wogonin concentration suggesting possible DNA intercalation of Wogonin. Further, in silico molecular docking of Wogonin on mammalian DNA also indicated possible intercalation of Wogonin with DNA. The denaturation and FRET studies revealed that Wogonin prevents denaturation of DNA strands and provide stability to genomic DNA against a variety of chemical denaturants. The cellular uptake study showed that Wogonin enters osteoarthritis chondrocytes and was mainly localized in the nucleus. Wogonin treatment to OA chondrocytes protects the fragmentation of genomic DNA in response to IL-1beta as evaluated by DNA ladder and TUNEL assay. Treatment of chondrocytes with Wogonin resulted in significant suppression of IL-1beta-mediated induction of ROS. Further, Wogonin exhibited protective potential through potent suppression of extrinsic and intrinsic apoptotic pathways and induction of anti-apoptotic proteins in
Khan Nazir M; Ahmad Imran; Ansari Mohammad Y; Haqqi Tariq M
Chemico-biological interactions
2017
2017-08
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1016/j.cbi.2017.06.025" target="_blank" rel="noreferrer noopener">10.1016/j.cbi.2017.06.025</a>
Wogonin, a plant derived small molecule, exerts potent anti-inflammatory and chondroprotective effects through the activation of ROS/ERK/Nrf2 signaling pathways in human Osteoarthritis chondrocytes.
*ERK1/2; *Nrf2; *Osteoarthritis; *Redox; *Wogonin; Anti-Inflammatory Agents/administration & dosage; Chondrocytes/drug effects/pathology; Flavanones/*administration & dosage; Gene Expression Regulation/drug effects; Humans; Inflammation/*drug therapy/metabolism/pathology; Kelch-Like ECH-Associated Protein 1/chemistry/*genetics/metabolism; MAP Kinase Signaling System/drug effects; Molecular Docking Simulation; NF-E2-Related Factor 2/antagonists & inhibitors/chemistry/*genetics/metabolism; Osteoarthritis/*drug therapy/metabolism/pathology; Protein Binding; Reactive Oxygen Species/metabolism; Signal Transduction/drug effects
Osteoarthritis (OA), characterized by progressive destruction of articular cartilage, is the most common form of human arthritis. Here, we evaluated the potential chondroprotective and anti-inflammatory effects of Wogonin, a naturally occurring flavonoid, in IL-1beta-stimulated human OA chondrocytes and cartilage explants. Wogonin completely suppressed the expression and production of inflammatory mediators including IL-6, COX-2, PGE2, iNOS and NO in
Khan Nazir M; Haseeb Abdul; Ansari Mohammad Y; Devarapalli Pratap; Haynie Sara; Haqqi Tariq M
Free radical biology & medicine
2017
2017-05
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1016/j.freeradbiomed.2017.02.041" target="_blank" rel="noreferrer noopener">10.1016/j.freeradbiomed.2017.02.041</a>
Zcchc6 and Zcchc11 exhibit cell autonomous effects on bone remodeling in mice.
Sondag G; Haynie S; Safadi F F; Haqqi T M
Osteoarthritis & Cartilage
2017
2017-04-02
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1016/j.joca.2017.02.041" target="_blank" rel="noreferrer noopener">10.1016/j.joca.2017.02.041</a>