牛磺酸对3T3-L1前脂肪细胞分化的影响

Effect of taurine on the differentiation of 3T3-L1 preadipocytes

目的:有研究发现,喂食牛磺酸的糖尿病鼠脂肪细胞摄糖的能力增加,但牛磺酸是否能影响脂肪细胞分化,目前还尚不清楚。本实验观察牛磺酸对3T3-L1前脂肪细胞分化的影响,并探讨牛磺酸影响3T3-L1前脂肪细胞分化的机制。方法:实验于2006-07/09在中南大学湘雅三医院实验中心完成。①实验材料:实验中应用的3T3-L1细胞由中国科学院上海细胞库提供。②实验方法:将3T3-L1细胞用含体积分数为0.10胎牛血清的高糖DMEM培养液培养,内含108u/L青霉素,80×1010U/L链霉素。细胞达汇片后,按5×107L-1密度接种于培养瓶中,应用0.5mmol/LIBMX、0.5mg/L胰岛素和1μmol/L地塞米松诱导分化,实验组加入牛磺酸,对照组未实施牛磺酸干预。油红O染色观察脂肪细胞分化情况。10,20mmol/L牛磺酸分别干预C3T3-L1前脂肪细胞24,48h,抽提实验组和对照组细胞RNA及蛋白。③实验评估:采用RT-PCR及Western-blot方法观察脂肪分化相关基因的表达变化。结果:①10mmol/L牛磺酸干预后14d,实验组油红O染色阳性脂肪细胞明显少于对照组。②10,20mmol/L牛磺酸分别干预3T3-L1前脂肪细胞24,48h后,对脂肪分化相关基因Insig-2蛋白、PPAR、Insing-2、脂联素、脂联素受体、GLUT-4、AP-2mRNA表达无明显影响。结论:牛磺酸可抑制前脂肪细胞分化,但具体机制还需要进一步研究。

AIM： Some studies found that the glucose uptake ability of diabetes mellitus mice is stronger after feeding with taurine. Whether the taurine can affect adipocytes or not deserves further studies. This article investigates the effects of taurine on the differentiation of 3T3-L1 preadipocytes and detects the mechanics. METHODS： Experiments were performed in the Laboratory Center of Xiangya Third Hospital of Central South University from July to September 2006. （1）3T3L1 preadipocytes were provided by Shanghai Cell Bank of China Academy of Sciences. （2）3T3-L1 preadipocytes were cultured in high glucose DMEM medium containing fetal bovine serum of 0.10 volume fraction, 10^8 u/L benzylpenicillin and 80 ×10^10 U/L streptaquaine. After confluence, cells at 5 × 10^7 L^-1 were incubated in culture flask, and induced with 0.5 mmol/L IBMX, 0.5 mg/L insulin and 1 μmol/L dexamethasone. The cells in an experimental group were intervened with taurine, whereas these in a control group were not treated with taurine. Oil O staining was used to observe the development of adipose cells. C3T3-L1 preadipocytes were treated with 10, 20 mmol/L taurine respectively for 24 and 48 hours. RNA and protein were extracted in the control group and the experimental group. （3）Genes related to adipose cells development was examined by RT-PCR and Western-blot. RESULTS： （1）The number of cells stained by oil O was less in the experimental group than the control group in 3T3-L1 preadipocytes treated with 10 mmol/L taurine for 14 days. （2）After C3T3-L1 preadipocytes were treated with 10, 20 mmol/L taurine respectively for 24 and 48 hours, there were no changes in expression of Insig-2 protein, PPAR, Insing-2, adiponectin, adiponectin receptor, GLUT-4, AP-2 mRNA. CONCLUSION： Taurine inhibits the differentiation of 3T3-L1 preadipocytes, but the mechanics still need to be investigated.