Preliminary study on mechanism of tricarboxylic acid cycle disruption in diabetic nephropathy cell model

Abstract

Abstract:

AIM: To explore the difference of tricarboxylic acid cycle (TCA) intermediates levels between diabetic nephropathy (DN) model cells and normal HK-2 cells and the metabolic disruption of DN model. METHODS: HK-2 cells were cultured in vitro and were divided into normal group, glucose group, palmitic acid(PA) group, PA high glucose group. Kidney cell damage was observed by fluorescence microscopy after mitochondrial staining. Metabolic disruption was evaluated by a GC-MS based metabolomics method. Concentration of TCA intermediates was measured by a GC-MS based quantification method. Expressions of relative enzymes were measured by qPCR method. RESULTS:High glucose did not exert as strong an effect on cell damage or metabolic disruption as PA, which caused obvious cell damage or metabolic disruption in a short period. The combined inducement presented the strongest effect. TCA intermediates variation was the main change in PA caused HK-2 cells metabolic disruption. Further, PA could increase the level of citrate and succinate in HK-2 cells. And instead of SDH in model cell, the increase of succinate level was closely related to succinate-CoA ligase. CONCLUSION: Metabolic disruption of TCA induced by PA may be closely related to diabetic nephropathy.

Key words: diabetic nephropathy, HK-2, palmitic acid, succinate

CLC Number:

R965.2

YU Xiaoyi, ZHOU Zhiruo, XIE Yuan, SUN Runbin, FEI Fei, GAO Haoxue, HUANG Jingqiu, A Jiye, WANG Guangji. Preliminary study on mechanism of tricarboxylic acid cycle disruption in diabetic nephropathy cell model[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2017, 22(5): 481-489.

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