摘要
目的观察小鼠过氧化物酶体增殖物激活受体γ1(PPARγ1)基因高表达对游离脂肪酸(FFA)介导的胰岛βTC3细胞损伤的影响。方法克隆构建重组载体pcDNA3.1/PPARγ1,并在βTC3细胞中进行稳定表达,用半定量RTPCR对其表达进行鉴定。之后,采用四唑盐(MTT)比色试验比较细胞暴露于较高水平FFA48h后的细胞活力。结果克隆出中国昆明小鼠PPARγ1全长基因,其cDNA序列与Genbank的PPARγ1基因序列基本相同,仅编码第421位天冬酰胺的密码子由AAU变成了AAC。经鉴定PPARγ1基因有效地在βTC3细胞中获得了表达。野生型βTC3细胞暴露于较高水平FFA48h后细胞活力下降(P<0.01),且FFA浓度越高细胞活力下降越明显(r=-0.962,P<0.01);而PPARγ1高表达βTC3细胞的细胞活力无明显改变(P>0.05)。结论PPARγ1高表达可保护胰岛βTC3细胞免于FFA介导的损伤。
To observe the protective effect of high expression of mouse proxisome proliferator activated receptor γ1 （PPARγ1） on free fatty acid （FFA）-induced βTC3 cell impairment. Methods The recombinant plasmid pcDNA3.1/PPARγ1 was generated with cloning and was stably transfected into pancreatic β TC3 cells. The expression was detected with semi-quantitative RT-PCR. Then the cell viability of wild βTC3 cells was compared with that of the βTC3 cells with high expressed PPARγ1 by MTT viability assay after they were exposed to high-level FFA for 48 hours. Results The sequencing results for amplified target gene showed that the sequence of PPARγ1 from Chinese Kunming mouse is similar to that of mouse PPARγ1 in Genebank, only the codon coding Asp at the site of 421 amino acid changed from AAU to AAC. PPARγ1 was efficiently expressed in βTC3 cells in vitro. The cell viability of wild βTC3 cells reduced after being exposed to high-level FFA for 48 hours（P＜0. 01）. Higher the level of FFA was, more obvious the reduction of the cell viability was （r=-0. 962, P＜0.01）. However, at the same condition, the cell viability of the βTC3 cells high expressing PPARγ1 had no significant change （P＞0.05）. Conclusion The high expression of PPARγ1 could protect βTC3 cells from FFA-induced impairment.
基金
福建省自然科学基金资助项目(C0410022)
