PKCδ在游离脂肪酸诱导内皮细胞凋亡过程中的作用研究

The investigation of the role of PKCδin free fatty acid-induced endothelial cell apoptosis

目的探讨蛋白激酶Cδ(PKCδ)在游离脂肪酸(FFAs)诱导内皮细胞凋亡过程中的作用。方法培养人脐静脉内皮细胞,分别给予不同浓度的FFAs刺激,转染PKCδsiRNA以抑制PKCδ的表达。使用比色法检测细胞的增殖情况,采用定量流式细胞术测定细胞凋亡情况,免疫印迹法检测PKCδ蛋白及磷酸化蛋白的表达水平。结果在人脐静脉内皮细胞内FFAs可产生多种效应,包括浓度依赖性的抑制细胞增殖、诱导细胞凋亡、增加PKCδ的表达和磷酸化等。抑制PKCδmRNA的表达可导致FFAs诱导细胞凋亡减少。结论 PKCδ可能介导内皮细胞中FFAs诱导的细胞凋亡。

Objective To investigate the role of PKCδ in free fatty acid-induced endothelial cell apoptosis. Methods In addi- tion, we looked for evidence of apoptosis-related PKCδ signal pathway. Human umbilical vein endothelial cells were treated with va- rious concentrations of free fatty acids and transiently transfected with PKCδ siRNA to inhibit PKCδ expression. Cell proliferation was determined through colorimetric assays,and apoptosis was quantified using flow cytometry. Protein expression was determined from cell lysates use Western blots with antibodies against p-PKCδ Tyr512,PKCδ. Statistical analyses were performed. Results Free fatty acids had multiple effects on human umbilical vein endothelial cells, including concentration-dependent inhibition of cell proliferation,induction of apoptosis, increased Fas expression, and increased PKCδ expression and phosphorylation. Inhibition of PKCδ mRNA expression by PKCδ siRNA led to a reduction in free fatty acid-induced apoptosis. Conclusion The free fatty acids- induced apoptosis in endothelial cells are possibly mediated by PKCS.