Modulating Of Chromium Gluconate On Cell Proliferation, Differentiation And Apoptosis In3T3-L1Adipocytes

Trivalent chromium are necessary trace elements in human and animal, as the component ofglucose tolerance factor, trivalent chromium is associated with glucose, fat and protein metabolism.Lack of trivalent chromium interfere with the metabolism of carbohydrases, and causehyperglycemia and hyperlipidemia, however, these symptoms can be alleviated by supplementingtrivalent chromium. It can promote fat burning and reduce body fat and weight. The main way tocurb obesity include reducing energy consumption, increasing energy expenditure, inhibiting3T3-Ll adipocytes differentiation and proliferation, lowering fat synthesis in fat cells, promotingfat decomposition and oxidation and so on.3T3-Ll adipocytes, as consecutive subculture cellsublimes, were isolated from3T3cells cloning. Cell has experienced the transformation of3T3-Lladipocytes into fat cells from the rapid proliferation to confluence and contact inhibition,3T3-Lladipocytes are used to for simulation research on body fat cells proliferation and differentiation.The main purposes of this study is to explore the effects of CrGlu and CrPic on cellproliferation, differentiation and apoptosis in3T3-Ll adipocytes by the vitro culture method. MTTassay, LDH assay, flow cytometry and fluorescence microscope were used for measuring cellgrowth inhibition (IC50), membrane permeability, cell apoptosis and the morphological features ofapoptotic cells, respectively. At the same time, it had carried on the effect of the differentconcentrations of trivalent chromium on lipid deposition efficiency of3T3-Ll adipocytes, and thepreliminary research on related genes. This study compared physiological activity differencebetween different ligands of trivalent chromium, and provided theoretical support for theapplication of trivalent chromium.The results were as follows:1. Trivalent chromium inhibited the proliferation of3T3-Ll adipocytes in a does-andtime-independent way. Compared to the untreated cells, after incubation with50,100,200and200μmol/L of trivalent chromium for48h, cell viability was reduced significantly, meanwhile it caninduced cell apoptosis. It was also found that200μmol/L CrGlu had higher physiological activityon inhibition of3T3-Ll adipocytes cell proliferation, compared with CrPic.2. When3T3-Ll adipocytes were treated with100~200μmol/L CrGlu, it can significantlyinhibited3T3-L1adipocytes differentiation, reduced intracellular lipid accumulation and promotedthe degradation of intracellular triglyceride. With the increase of the concentration of the CrGlu,its physiological activity is stronger; Low concentration (10~50μmol/L) CrGlu had no effect on 3T3-L1adipocytes differentiation. It was also found that200μmol/L CrGlu had higherphysiological activity on inhibition of3T3-Ll adipocytes cell differentiation, compared with CrPic.3. Trivalent chromium can inhibited the activity of PPARγ and C/EBPα mRNA, meanwhilelowered PPARγ and C/EBPα mRNA and protein expression, and it showed that higherconcentrations, stronger inhibition activity; It was also found that200μmol/L CrGlu had higherphysiological activity on inhibition of PPARγ and C/EBPα mRNA and protein expression,compared with CrPic. We also found that the low concentration (10~50μmol/L) CrGlu had noobvious effect on PPARγ and C/EBPα mRNA expression content(P＜0.05).Through the above results we can arrive at some preliminary conclusions: Trivalent chromiumfrom had an inhibitory effect on3T3-Ll adipocytes in vitro, and could induce apoptosis; The studyalso confirmed that high concentrations of CrGlu (100~200μmol/L) can inhibit adipocytedifferentiation, reduce intracellular lipid accumulation. Under the same concentration (200μmol/L), CrGlu compared with CrPic has higher inhibition efficiency of intracellular lipidaccumulation, further study showed that in the process, can significantly reduced PPARγ andC/EBPα mRNA expression.