论文部分内容阅读
目的:通过RNA干扰技术特异性抑制LITAF基因,观察HPA-v人前脂肪细胞胰岛素信号通路的变化情况。方法:将HPA-v人前脂肪细胞和THP-1源巨噬细胞共培养,分为3组,分别为阴性对照组(Scramble组)、siRNA组和正常对照组(NC组),通过qRT-PCR,Western blot法检测LITAF,IRS-2,p-PKC,PI3-K及GLUT2的表达。结果:在软脂酸的刺激下Scramble组LITAF的表达较高于LITAF siRNA和IVC组;IRS-2,p-PKC,PI3-K及GLUT2在LITAF RNAi组较NC组和Scramble组明显提高(P<0.05)。结论:LITAF参与了软脂酸引起的脂肪细胞胰岛素抵抗,而脂肪酸作用于巨噬细胞的受体TLR4同样也是LITAF通路的细胞膜受体,软脂酸减弱了对胰岛素信号通路分子的抑制作用,因此LITAF参与胰岛素抵抗对糖尿病的发病起了至关重要的作用。
OBJECTIVE: To observe the changes of insulin signaling pathway in HPA-v human preadipocytes by specific inhibition of LITAF gene by RNA interference technology. METHODS: HPA-v human preadipocytes and THP-1-derived macrophages were co-cultured and divided into three groups: negative control group (Scramble group), siRNA group and normal control group (NC group) The expressions of LITAF, IRS-2, p-PKC, PI3-K and GLUT2 were detected by Western blot. RESULTS: The expression of LITAF in Scramble group was higher than that in LITAF siRNA and IVC group under the stimulation of palmitate. The levels of IRS-2, p-PKC, PI3-K and GLUT2 in LITAF RNAi group were significantly higher than those in NC group and Scramble group <0.05). CONCLUSION: LITAF is involved in the insulin resistance of adipocytes induced by palmitate, and TLR4, a receptor for fatty acids acting on macrophages, is also a cell membrane receptor of the LITAF pathway. Palmitate weakens the inhibitory effect on insulin signaling molecules and therefore LITAF participation in insulin resistance plays a crucial role in the pathogenesis of diabetes.