软脂酸对人脐静脉内皮细胞PI3K/Akt表达的影响及六味地黄丸的干预作用

Effect of palmitic acid on the expression of PI3K/Akt in human umbilical vein endothelial cells and the intervention of liuweidihuang pill

目的:研究软脂酸(palmitic acid,PA)对人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVEC)PI3K/Akt表达的影响及六味地黄丸(liuweidihuang pill,LWDHP)含药血清的干预效应。方法:首先制备LWDHP含药血清,然后将培养的HUVEC作为研究对象,分为正常对照组,模型组,5%LWDHP含药血清治疗组,二甲双胍(metformin,MET)组,药物处理后采用RT-PCR及Western blot检测细胞内PI3K/Akt的m RNA和蛋白的表达。结果:RT-PCR结果显示模型组细胞的PI3K/Akt的m RNA表达量较正常对照组显著减少(PPI3K=0.000,PAkt=0.000),5%LWDHP含药血清组(PPI3K=0.012,PAkt=0.000)及MET组(PPI3K=0.000,PAkt=0.000)细胞内的PI3K/Akt m RNA表达量较模型组增加,5%LWDHP含药血清组细胞内PI3K/Akt m RNA表达量与MET组比较有统计学意义(PPI3K=0.000,PAkt=0.000)。同时Western blot结果显示模型组细胞的PI3K/P-Akt的蛋白表达较正常对照组也显著减少(PPI3K=0.000,PP-Akt=0.000),5%LWDHP含药血清组(PPI3K=0.000,PP-Akt=0.000)和MET组(PPI3K=0.000,PP-Akt=0.000)细胞内PI3K/P-Akt蛋白表达量较模型组同样明显增加,5%LWDHP含药血清组细胞内PI3K/P-Akt蛋白表达量与MET组比较有统计学意义(PPI3K=0.003,PP-Akt=0.001)。结论:LWDHP干预保护后可以促进PA诱导损伤的HUVEC内PI3K/Akt m RNA和蛋白的表达。

Objective：To observe the effect of palmitic acid（PA） on the expression of PI3K/Akt m RNA and protein in human umbilical vein endothelial cells（HUVEC） and the intervention of the serum containing liuweidihuang pill（LWDHP）. Methods：Firstly the serum containing LWDHP was prepared,and then the human umbilical vein endothelial cells were divided into normal group,model group,LWDHP group with serum containing concentration of 5% and metformin（MET） group. The m RNA and protein expression levels of PI3K/Akt were determined by RT-PCR and Western blot,respectively. Results ：RT-PCR showed that m RNA expression of PI3K/Akt was significantly lower in model group than in normal group（PPI3K=0.000,PAkt=0.000）,while m RNA expression of PI3K/Akt was significantly higher in 5% LWDHP group（PPI3K=0.012,PAkt=0.000） and in MET group（PPI3K=0.000,PAkt=0.000） than in model group.There were statistically significant differences in m RNA expression of PI3K/Akt between MET group and 5% LWDHP group（PPI3K=0.000,PAkt=0.000）. Western blot showed that protein expression of PI3K/P-Akt was also significantly lower in model group than in normal group（PPI3K=0.000,PP-Akt=0.000）,while protein expression of PI3K/P-Akt was also significantly higher in 5% LWDHP group（PPI3K=0.000,PP-Akt=0.000） and in MET group（PPI3K=0.000,PP-Akt=0.000） than in model group. There were statistically significant differences in protein expression of PI3K/Akt between MET group and 5%LWDHP group（PPI3K=0.003,PP-Akt=0.001）. Conclusion ：The serum containing LWDHP could promote the expression of PI3K/Akt m RNA and protein in HUVEC damaged by PA.