Abstract
Taurochenodeoxycholic acid (TCDCA) is one of the main components of bile acids (BAs). TCDCA has been reported as a signaling molecule, exerting anti-inflammatory and immunomodulatory functions. However, it is not well known whether those effects are mediated by TGR5. This study aimed to elucidate the interaction between TCDCA and TGR5. To achieve this aim, first, the TGR5 eukaryotic vector was constructed. The expression level of TGR5 in 293T cells was determined by immunofluorescence, real-time quantitative PCR (RT-PCR, qPCR), and Western blot. The luciferase assay, fluorescence microscopy, and enzyme-linked immunosorbent assay (ELISA) were recruited to check the interaction of TCDCA with TGR5. TCDCA treatment in 293T cells resulted in TGR5 internalization coupled with a significant increase in cAMP luciferase expression. Our results demonstrated that TCDCA was able to bind to the TGR5 receptor and activate it. These results provide an excellent potential therapeutic target for TCDCA research. Moreover, these findings also provide theoretical evidence for further TCDCA research.
Highlights
BAs are major components of bile and can be divided into free bile acids and conjugated bile acids based on their chemical structure
TGR5 targeted gene was cloned from pMD19-T-TGR5 plasmid, which acted as a pair of primers cloned to pCMV-EGFP plasmid tested by the double enzyme digestion and comparison on the GeneBank
We found that the cAMP content was augmented in the 293T cells that had been treated with 1 μM Taurochenodeoxycholic acid (TCDCA), and it continued to increase in the 100 μM of TCDCA
Summary
BAs are major components of bile and can be divided into free bile acids and conjugated bile acids based on their chemical structure. Free bile acids include cholic acid (CA), deoxycholic acid (DCA), chenodeoxycholic acid (CDCA), and lithocholic acid (LCA). Conjugated bile acids include glycocholic acid (GCA), taurocholic acids (TCA), glycochenodeoxycholic acid (GCDCA), and TCDCA [1]. BAs are amphipathic molecular, meaning that they can prevent precipitation of cholesterol crystals, decreasing cholesterol stone formation [4,5]. BAs have another function in anti-inflammation and immunology. Current evidence suggests that ursodeoxycholic acid (UDCA) can provide multiple benefits, including the decreased hydrophobicity of the bile acid pool, increased hepatobiliary secretion, reduced inflammation, and cell death [7]. In 2011, Yusuke Iguchi and colleagues reported that UDCA activated TGR5, providing significant evidence and a method to continue the research on UDCA and other BAs [8]
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