RNA interference for CFTR attenuates lung fluid absorption at birth in rats.
Female; Pregnancy; Rats; Genes; Cells; Animal Population Groups; Animal Studies; Lung; Respiratory Mucosa; Genes – Drug Effects; Biological Transport – Drug Effects; Lung – Drug Effects; Lung – Metabolism; Membrane Proteins – Metabolism; Respiratory Mucosa – Drug Effects; Respiratory Mucosa – Metabolism; RNA – Metabolism; RNA – Pharmacodynamics; Water – Metabolism
Background: Small interfering RNA (siRNA) against αENaC (α-subunit of the epithelial Na channel) and CFTR (cystic fibrosis transmembrane conductance regulator) was used to explore ENaC and CTFR function in newborn rat lungs. Methods: Twenty-four hours after trans-thoracic intrapulmonary (ttip) injection of siRNAgenerating plasmid DNA (pSi-0, pSi-4, or pSi-C2), we measured CFTR and ENaC expression, extravascular lung water, and mortality. Results: αENaC and CFTR mRNA and protein decreased by \textasciitilde80% and \textasciitilde85%, respectively, following αENaC and CFTR silencing. Extravascular lung water and mortality increased after αENaC and CFTR-silencing. In pSi-C2-transfected isolated DLE cells there were attenuated CFTR mRNA and protein. In pSi-4-transfected DLE cells αENaC mRNA and protein were both reduced. Interestingly, CFTR-silencing also reduced αENaC mRNA and protein. αENaC silencing, on the other hand, only slightly reduced CFTR mRNA and protein. Conclusion: Thus, ENaC and CFTR are both involved in the fluid secretion to absorption conversion around at birth.
Li T; Koshy S; Folkesson H G
Respiratory research
2008
2008-01
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/10.1186/1465-9921-9-55" target="_blank" rel="noreferrer noopener">10.1186/1465-9921-9-55</a>
RNA interference for CFTR attenuates lung fluid absorption at birth in rats.
*RNA Interference; Animals; Biological Transport/drug effects; Cells; Cultured; Cystic Fibrosis Transmembrane Conductance Regulator/*genetics/*metabolism; Epithelial Sodium Channels/genetics/metabolism; Female; Gene Expression Regulation/drug effects; Lung/cytology/drug effects/*metabolism; Messenger/metabolism; Newborn; Pregnancy; Rats; Respiratory Mucosa/cytology/drug effects/metabolism; RNA; Small Interfering/*pharmacology; Sprague-Dawley; Water/*metabolism
BACKGROUND: Small interfering RNA (siRNA) against alphaENaC (alpha-subunit of the epithelial Na channel) and CFTR (cystic fibrosis transmembrane conductance regulator) was used to explore ENaC and CTFR function in newborn rat lungs. METHODS: Twenty-four hours after trans-thoracic intrapulmonary (ttip) injection of siRNA-generating plasmid DNA (pSi-0, pSi-4, or pSi-C2), we measured CFTR and ENaC expression, extravascular lung water, and mortality. RESULTS: alphaENaC and CFTR mRNA and protein decreased by approximately 80% and approximately 85%, respectively, following alphaENaC and CFTR silencing. Extravascular lung water and mortality increased after alphaENaC and CFTR-silencing. In pSi-C2-transfected isolated DLE cells there were attenuated CFTR mRNA and protein. In pSi-4-transfected DLE cells alphaENaC mRNA and protein were both reduced. Interestingly, CFTR-silencing also reduced alphaENaC mRNA and protein. alphaENaC silencing, on the other hand, only slightly reduced CFTR mRNA and protein. CONCLUSION: Thus, ENaC and CFTR are both involved in the fluid secretion to absorption conversion around at birth.
Li Tianbo; Koshy Shyny; Folkesson Hans G
Respiratory research
2008
2008-07
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/10.1186/1465-9921-9-55" target="_blank" rel="noreferrer noopener">10.1186/1465-9921-9-55</a>
Stimulation of MAP kinase pathways after maternal IL-1beta exposure induces fetal lung fluid absorption in guinea pigs.
Absorption/drug effects; Animal/*metabolism; Animals; Body Fluids/*metabolism; Extracellular Signal-Regulated MAP Kinases/metabolism; Female; Fetus/metabolism; Gestational Age; Guinea Pigs; Hydrocortisone/*metabolism; Injections; Interleukin-1beta/administration & dosage/*pharmacology; Lung/*embryology; Mitogen-Activated Protein Kinase Kinases/metabolism; Mitogen-Activated Protein Kinases/*metabolism; Pregnancy; Rats; Recombinant Proteins/administration & dosage/pharmacology; Subcutaneous
BACKGROUND: We tested the hypothesis that maternal interleukin-1beta (IL-1beta) pretreatment and induction of fetal cortisol synthesis activates MAP kinases and thereby affects lung fluid absorption in preterm guinea pigs. METHODS: IL-1beta was administered subcutaneously daily to timed-pregnant guinea pigs for three days. Fetuses were obtained by abdominal hysterotomy and instilled with isosmolar 5% albumin into the lungs and lung fluid movement was measured over 1 h by mass balance. MAP kinase expression was measured by western blot. RESULTS: Lung fluid absorption was induced at 61 days (D) gestation and stimulated at 68D gestation by IL-1beta. Maternal IL-1beta pretreatment upregulated ERK and upstream MEK expression at both 61 and 68D gestation, albeit being much more pronounced at 61D gestation. U0126 instillation completely blocked IL-1beta-induced lung fluid absorption as well as IL-1beta-induced/stimulated ERK expression. Cortisol synthesis inhibition by metyrapone attenuated ERK expression and lung fluid absorption in IL-1beta-pretreated fetal lungs. JNK expression after maternal
Bhattacharjee Reshma; Li Tianbo; Koshy Shyny; Beard LaMonta L; Sharma Kapil; Carter Ethan P; Garat Chrystelle; Folkesson Hans G
Respiratory research
2007
2007-03
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/10.1186/1465-9921-8-27" target="_blank" rel="noreferrer noopener">10.1186/1465-9921-8-27</a>