二甲双胍对兔动脉粥样硬化血管平滑肌细胞增殖的影响

Effects of metformin on the e proliferation of vascular smooth muscle cells in rabbit atherosclerosis model

目的研究二甲双胍对兔动脉粥样硬化过氧化物酶体增殖物激活受体γ(PPARγ)、细胞间黏附分子-1(ICAM-1)的表达及血管平滑肌细胞(VSMCs)增殖的影响。方法按照体重将新西兰兔随机分为4组:正常组、正常给药组、模型组和模型给药组,每组10只。模型组和模型给药组通过高脂饮食+主动脉内膜损伤术建立动脉粥样硬化模型。造模成功后,模型给药组和正常给药组均给予二甲双胍100 mg·kg^-1 d^-1;正常组和模型组给予0.9%NaCl,均灌胃8周。以免疫组织化学染色和实时荧光定量-PCR分别检测PPARγ和ICAM^-1的蛋白和基因表达水平,以四甲基氮唑蓝法测定细胞增殖水平(OD值)。结果正常组、正常给药组、模型组和模型给药组PPARγ蛋白阳性表达率分别为12.67%,10.80%,60.32%和21.49%;这4组的ICAM^-1蛋白阳性表达率分别为20.80%,19.01%,68.63%和43.24%;这4组的PPARγ基因的相对表达量分别为1,0.96±0.28,4.87±1.23和2.39±0.68;这4组的ICAM^-1基因的相对表达量分别为1,0.92±0.31,6.18±1.35和3.75±1.04;这4组的VSMCs细胞增殖水平分别为1,0.94±0.27,4.23±1.59和2.37±0.54。上述指标:模型组和模型给药组与正常组和正常给药组相比,差异均有统计学意义(均P<0.05);模型给药组与模型组相比,差异均有统计学意义(均P<0.05)。结论二甲双胍对兔动脉粥样硬化PPARγ和ICAM^-1的蛋白和基因表达以及血管平滑肌细胞增殖活性具有抑制作用。

Objective To investigate the effects of metformin on the expression of peroxisome proliferator-activated receptor γ(PPARγ),intercellular adhesion molecule-1(ICAM-1)and proliferation of vascular smooth muscle cells(VSMCs)in rabbit atherosclerosis model.Methods New Zealand rabbits were randomly divided into four groups according by weight:normal group,normal+drugs group,model group,model+drugs group,10 rabbits for each group.The model group and model+drugs group establishedcoronary atherosclerosis by high-fat diet and aortic intima damage atherosclerosis.While the normal+drugs group and model+drugs group were gavaged 100 mg·kg^-1 metformin group for 8 weeks,the other two groups officer give equal 0.9%NaCl.The protein and gene expression levels of PPARγand ICAM-1 were detected by immunohistochemistry and RT-PCR,respectively.Rabbit VSMCs proliferation(OD value)were detected by methyl thiazolyl tetrazolium assay.Results The positive expression rates of PPAR gamma protein in normal group,normal+drugs group,model group,model+drugs group were 12.67%,10.80%,60.32% and 21.49%,respectively;the positive expression rate of ICAM-1 protein in the four groups were 20.80%,19.01%,68.63% and 43.24%,respectively;the relative expression levels of PPAR gamma mRNA in the four groups were 1,0.96±0.28,4.87±1.23 and2.39±0.68,respectively;the relative expression levels of ICAM-1 mRNA in the four groups were 1,0.92±0.31,6.18±1.35,3.75±1.04,respectively;the proliferation levels of VSMCs cells in the four groups were 1,0.94±0.27,4.23±1.59 and 2.37±0.54,respectively.Comparison between model group,model+drugs group and normal group,normal+drugs group,or comparison between model+drugs group and model group,the difference of the factors were significantly(all P<0.05).Conclusion Metformin could inhibit the protein and gene expression of PPARγand ICAM^-1 and the proliferation of VSMCs in rabbit atherosclerosis model.