辛伐他汀对高糖损伤血管内皮细胞的保护作用研究

The effect of simvastatin on high glucose induced vascular endothelial cells injury

目的观察辛伐他汀对高糖损伤血管内皮细胞的保护作用,并探讨其作用的机制。方法实验分为4组:对照组(健康体检者,A组),高血糖组(B组,33.3mmol/L葡萄糖),高血糖+低浓度辛伐他汀组(C组,33.3mmol/L葡萄糖+1.0μmol/L辛伐他汀),高血糖+高浓度辛伐他汀组(D组,33.3mmol/L葡萄糖+10.0μmol/L辛伐他汀)。细胞增殖使用CCK-8实验检测,细胞凋亡使用TUNEL检测法,蛋白表达使用Western blot方法检测,基因检测采用实时荧光定量PCR(real time PCR)。结果 B、C、D组的内皮细胞存活率分别为(42.5±6.4)%、(58.6±7.8%)和(71.3±11.7)%,各组之间差异有统计学意义(P<0.05)。A、B、C、D组内皮细胞的凋亡率分别为(1.8±0.6)%、(45.8±8.9)%、(22.7±6.4)%和(12.6±4.2)%,各组之间差异有统计学意义(P<0.05)。B组(0.13±0.03)内皮细胞Bcl-2蛋白的表达明显低于A组(1.02±0.16),C组(0.28±0.04)明显高于B组,D组(0.68±0.11)又明显高于C组(P<0.05,P<0.01)。B组内皮细胞bax基因的表达明显高于A组,C组明显低于B组,D组又明显低于C组(P<0.05,P<0.01)。结论辛伐他汀可以明显抑制高糖诱导的内皮细胞凋亡,上调Bcl-2蛋白表达和下调bax基因表达可能是其主要的作用机制。

Objective To study the effect of simvastatin on high glucose induced vascular endothelial cells injury and explore its action mechanism .Methods Endothelial cells were devided into four group：control group（A） ,high glucose（33 .3 mmol/L）group （B） ,high glucose（33 .3 mmol/L）＋simvastatin（1 .0 μmol/L）group（C） ,and high glucose（33 .3 mmol/L）＋simvastatin（10.0 μmol/L）group（D） .The inhibition of cells was detected by CCK-8 assay .The apoptosis was determined by TUNEL assay .The protein ex-pression was detected by Western blot analysis .Results The cells survival rates were （42 .5 ± 6 .4% ） ,（58 .6 ± 7 .8% ） ,and （71 .3 ± 11 .7% ） in B ,C ,and D group respectively .There was significant difference among the three groups（P〈0.05） .The per-cents of apoptosis were （1 .8 ± 0.6% ） ,（45 .8 ± 8 .9% ） ,（22 .7 ± 6 .4% ） and （12 .6 ± 4 .2% ） in A ,B ,C and D group ,respectively . There was significant difference among the four groups（P〈0.05） .The protein expression of Bcl-2 was lower in B group（0.13 ± 0. 03）than that in A group（1 .02 ± 0.16） .The Bcl-2 expression was higher in C group（0.28 ± 0.04）than that in B group ,and in D group（0.68 ± 0.11）than that in C group（P〈0.05 ,P〈0.01） .The gene expression of bax was higher in B group than that in A group .The bax expression was lower in C group than that in B group ,and the bax expression was lower in D group than that in C groups（P〈0.05 ,P〈0.01） .Conclusion Simvastatin can inhibit high glucose induced vascular endothelial cells apoptosis .Upregu-lation of Bcl-2 and downregulation of bax may be its action mechanism .