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α- ketoglutarate receptor activation hGPCRc Effects on cells of diabetic nephropathy model

Author: HuangLinYi
Tutor: LiuYongXue
School: PLA Military Academy of Medical Sciences
Course: Pharmacology
Keywords: hGPCRc α- ketoglutarate sodium Diabetic nephropathy Tubular epithelial cells Mesangial cells Cytokines
CLC: R96
Type: Master's thesis
Year: 2007
Downloads: 43
Quote: 0
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Abstract


In a preliminary study on hGPCRc, we dropped from a human colon tissue grams and was named a member of the receptor hGPCRc, and its tissue distribution, subcellular localization and possible ligands tested. 2004 has been reported hGPCRc endogenous ligand for the α-ketoglutarate, which results in the traditional sense of the nature of GPCR ligands far, it is prepared surprise. In this study, stable cell lines expressing hGPCRc engineering, and as cells detect α-ketoglutarate sodium activation after hGPCRc intracellular calcium ion concentration, and then create an in vitro diabetic nephropathy (Diabetic nephropathy, DN) model preliminarily discussed α-ketoglutarate receptor and the possible relationship between DN. The main results are as follows: a, α-ketoglutarate receptor is activated successfully established the biochemical effects of CHO-hGPCRc engineered cell lines, RT-PCR results showed that the resulting engineered cell lines stably expressing CHO-hGPCRc hGPCRc; α - ketoglutarate sodium can cause cell line engineering intracellular calcium ion concentration, expressed as calcium volatility increased significantly and dose-dependent manner; and α-ketoglutarate sodium induced intracellular calcium fluctuations, not by extracellular calcium concentration effect is the result of mobilization of cellular calcium stores. Two, α-ketoglutarate receptors on the DN model tubular epithelial cells (RTEC) of primary cultured renal tubular epithelial cells, and in 25mM established under high glucose conditions DN model. RT-PCR results showed that compared with normal cells RTEC, DN model intracellular TGF-β, α-SMA, CTGF, MCP-1 and ICAM-1, and many other cytokine mRNA expression levels were significantly increased; α-amyl ketone two sodium can upregulate the expression of hGPCRc, and the DN model cells increased mRNA expression of cytokines inhibited. Three, α-ketoglutarate DN model cell receptors on mesangial cells (MC) of primary cultured mesangial cells, and in 25mM established under high glucose conditions DN model. MTT results showed that, α-ketoglutarate sodium inhibits high glucose-induced MCs proliferation; RT-PCR results showed that, α-ketoglutarate sodium induced by high glucose can be lowered under TGF-β/MCP-1 mRNA expression increased. Conclusion: 1, α-ketoglutarate receptor hGPCRc be activated sodium α-ketoglutarate, and through the increased release of intracellular calcium stores intracellular calcium ion concentration. 2, the first time that α-ketoglutarate sodium DN model can adjust a variety of renal tubular epithelial cell cytokine mRNA expression changes, can inhibit renal tubular epithelial cells DN model myofibroblast transdifferentiation (TEMT) role and extracellular matrix (EMC) formation. 3, α-ketoglutarate sodium inhibits proliferation of mesangial cells and reducing DN model intracellular TGF-β, MCP-1 secretion.

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