高糖高脂、PAF对肾小球系膜细胞PKCβI mRNA表达的影响及阿托伐他汀干预作用观察

摘要

目的 观察阿托伐他汀对高糖高脂、血小板活化因子(PAF)诱导下蛋白激酶C-βI(PKCβI)mRNA表达的影响.方法 取培养好的、分化成熟的人肾小球系膜细胞(HMCs)细胞分为A正常对照组、B高糖高脂+PAF组、C高糖高脂+PAF+PKCβI抑制剂LY333531组、D高糖高脂+PAF+阿托伐他汀组共4个组.A组加入5.5mmol/L的D-葡萄糖;B组加入30 mmol/L的D-葡糖糖、20 mg/L的溶血磷脂酰胆碱(LPC)和2×10-8 mol/L的PAF C-16;C组经2×10-7 mol/L的LY333531预处理1 h后,加入30 mmol/L的D-葡萄糖、20 mg/L的LPC、2×10-8 mol/L的PAF C-16;D组经10μmol/L的阿托伐他汀处理1 h后,加入30 mmol/L的D-葡萄糖、20 mg/L的LPC、2×10-8 mol/L的PAF C-16.各组细胞培养24 h后,提取细胞RNA.采用实时荧光定量PCR法检测细胞中的PKCβI mRNA.结果 A、B、C、D组细胞中PKCβImRNA相对表达量分别为1.00±0.00、3.59±0.41、2.76±0.57、1.02±0.00.B组细胞中PKCβI mRNA相对表达量高于其余三组,D组PKCβI mRNA相对表达量低于B组和C组(P均<0.05).结论 高糖、高脂、PAF诱导下肾小球系膜细胞中PKCβI mRNA表达上调,阿托伐他汀作用后可抑制PKCβI mRNA表达.%Objective To investigate the effect of atorvastatin on the PKCβI mRNA expression of high glucose and lysophosphatidylcholine (LPC)and platelet activating factor (PAF)-induced mesangial cells. Methods Human glomeru-lar mesangial cells (HMCs)were divided into four groups:the normal control group (group A),high glucose and LPC +PAF group (group B),high glucose and LPC + PAF + LY333531 group (group C),and high glucose and high LPC +PAF + atorvastatin group (group D). We added 5. 5 mmol/ L D-glucose to group A;group B was treated with 30 mmol/ L D-glucose,20 mg/ L LPC,and 2 × 10 - 8 mol/ L PAF C-16;group C was pretreated with 2 × 10 - 7 mol/ L LY333531 for 1 h, and then was added with 30 mmol/ L D-glucose,20 mg/ L LPC,and 2 × 10 - 8 mol/ L PAF C-16;group D was pretreated with 10 μmol/ L atorvastatin for 1 h,and then was added with 30 mmol/ L D-glucose,20 mg/ L LPC,2 × 10 - 8 mol/ L PAF C-16. At 24 h after culture,RNA was extracted. The expression of PKCβI mRNA in the mesangial cells was detected by real-time fluorescence quantitative PCR. Results The relative expression of PKCβI-mRNA in the groups A,B,C,and D was 1. 00 ± 0. 00,3. 59 ± 0. 41,2. 76 ± 0. 57,and 1. 02 ± 0. 00,respectively. The expression of PKCβI-mRNA in the group B group was higher than that of the other three groups,and the expression of PKCβI-mRNA in the group D was sig-nificantly lower than that of the groups B and C (all P < 0. 05). Conclusions The expression of PKCβI mRNA in high glucose and LPC and PAF-induced glomerular mesangial cells is up-regulated,and atorvastatin can inhibit the PKCβI ex-pression.