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Studies On Interacation Of TCDCA And TGR5

Posted on:2015-11-22Degree:MasterType:Thesis
Country:ChinaCandidate:L K ShiFull Text:PDF
GTID:2323330536950423Subject:Basic veterinary science
Abstract/Summary:PDF Full Text Request
Taurochenodeoxycholic acid(TCDCA) is a main component of bile acids, and it has been reported that as signalingmolecules, it can regulate anti-inflammatory and immunomodilatory through a variety of signaling pathways mediated by TGR5. This study aimed is to research whether TCDCA can active TGR5 and mediate the signaling pathways.This study based on construced a TGR5 eukaryotic expression vector which expressed in 293 T cells. The expression of TGR5 were determine by immunofluorescence, real-time quantitative PCR and western blotting in 293 T cell,respectively. Receptor internalization wsa observed by fluorescence microscope; ElISA method was used to determine the level of intracellular cAMP, and analyse the dose-effect relationship of TCDCA and CRE-driven luciferase reporter in 239 T cells.The results show that TGR5 eukaryotic expression vector was constructed successfully and expressed in 293 T cells. After treatment with TCDCA, TGR5 was internalized, cAMP and luciferase reporter were increased significantly(p<0.01) in a dose- dependent manner(r2=0.9958) in 293 T cells which was transfected with TGR5.Conclusions: TCDCA can bind with TGR5.
Keywords/Search Tags:239T cell, TGR5, TCDCA, Internalization, c AMP, Luciferase reporter activity, Interaction
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