吡格列酮对游离脂肪酸作用下βTC-3细胞GPR40表达的干预作用

Effects of Pioglitazone on the Expression of βTC-3 Cell GPR40 Induced by Free Fatty Acids

目的探讨游离脂肪酸(FFA)作用下胰岛βTC-3细胞G蛋白受体40(GPR40)mRNA表达的改变以及吡格列酮(Piog)对此改变的干预作用。方法以βTC-3细胞为研究对象,分为对照组及FFA组(0.25,0.5及1mmol/L)。半定量RT-PCR方法检测GPR40的表达。用不同浓度的Piog(0,0.1,1,10μmol/L)预孵育βTC-3细胞6h,加入1mmol/LFFA继续孵育24h,半定量RT-PCR方法检测GPR40的表达。结果(1)FFA孵育12h后,各组间GPR40的表达差别无统计学意义。(2)FFA孵育24h后,与对照组比较,0.25mmol/LFFA组GPR40的表达无差异,但0.5mmol/L和1mmol/LFFA组GPR40表达下调(P<0.05);0.5mmol/L与1mmol/LFFA组比较GPR40表达差别无统计学意义。(3)与1mmol/LFFA组比较,0.1μmol/LPiog+1mmol/LFFA组GPR40mRNA表达无差异,而1μmol/LPiog+1mmol/LFFA组和10μmol/L+1mmol/LFFA组GPR40表达升高(P<0.01)。结论长期高浓度FFA作用能够下调βTC-3细胞GPR40的表达,而Piog有助于保护或减轻FFA水平异常导致的βTC-3细胞GPR40表达的损害。

Objective To investigate gene expressions of GPR40 inβTC-3 cells induced by free fatty acids（FFA） and the intervention effect of pioglitazone（Piog） on those expressions. Methods βTC-3 cells studied in the experiment were subject to different treatments with a blank and three concentrations of FFA （0.25, 0.5 and 1.0 mmol/L）. Semi-quantitative RT-PCR analysis was used to determine the RNA expression level of GPR40 in βTC-3 cells after 12 hours and 24 hours respectively. Afterwards, βTC-3 cells were preincubated with different concentrations of Piog（0, 0. 1, 1, 10 μmol/L）for 6 hours, and 1 mmol/L FFA was then added and the cells were incubated for another 24 hours. The RNA expression level of GPR40 was determined by semi-quantitative RT-PCR at last. Results （1） After the cells were incubated with FFA of different concentrations for 12 hours, there was no statistically significant difference among all the groups. （2） After the 24-hour incubation, no difference was shown between the group with 0.25 mmol/L FFA and the control group. But compared to the control group and the group with 0.25 mmol/L FFA, the expression of GPR40 in βTC-3 cells in both groups with 0.5 mmol/L FFA and that with 1 mmol/L FFA decreased after the 24-hour incubation （P〈0.05）. There was, however, no statistical difference between the group with 0.5 mmol/L FFA and that with 1 mmol/L FFA. （3） Though no statistically significant difference was found between the group with 0. 1 /μmol/L Piog＋ 1.0 mmol/L FFA and that with 1.0 mmol/L FFA, the RNA expression level of GPR40 in both the group with 1 μmol/L Piog＋1. 0 mmol/LFFA and that with 10 μmol/L Piog＋1. 0 mmol/L FFA increased compared to the group with 1 mmol/L FFA （P〈0.01）. Conclusion The expression of GPR40 in βTC-3 cells may be inhibited by long-term administration of FFA of high concentration, while Piog can help to protect the expression of GPR40 against FFA-induced impairment in βTC-3 cell.