葛根素对兔动脉粥样硬化基质金属蛋白酶-9及其组织抑制物-1表达的影响

Effect of puerarin on expression of matrix metalloproteinase-9 and matrix metalloproteinase tissue ihibitor-1 in rabbit with atherosclerosis

目的:观察葛根素对动脉粥样硬化兔髂动脉分泌和表达基质金属蛋白酶-9(MMP-9)及其组织抑制物-1(TIMP-1)的影响。方法:20只家兔分为正常对照组(正常饮食,6只)、病理对照组(球囊和高脂饮食,8只)和葛根素组(球囊、高脂饮食和葛根素,8只)。球囊损伤后4周处死,取一侧病变髂动脉做病理切片,应用免疫组化法测定MMP-9和TIMP-1的蛋白表达;取另一侧病变髂动脉抽提总RNA应用半定量逆转录多聚酶链式反应(RT-PCR)测定MMP-9和TIMP-1 mRNA的表达。结果:兔动脉粥样斑块MMP-9 mRNA(mRNA/GAP-DH mRNA)表达:正常对照组、病理对照组、葛根素组的分别为0.81±0.17,1.52±0.24,1.03±0.19,病理对照组、葛根素组的较正常对照组显著增加(P<0.05),而葛根素组的较病理对照组显著下降(P<0.05),上述三组的TIMP-1 mRNA的表达依次为1.44±0.14,2.63±0.16,2.67±0.12,病理对照组与葛根素组的较正常对照组显著增加(P<0.05),但病理对照组与葛根素组间无显著差异(P>0.05)。免疫组化检测显示葛根素抑制MMP-9蛋白质表达(P<0.05),但对TIMP-1蛋白质的表达无影响。结论:葛根素可能是通过调节兔动脉粥样斑块分泌MMP-9途径发挥稳定动脉粥样硬化斑块的作用。

Objective： To investigate the effect of puerarin on expression of matrix metalloproteinase-9 （MMP-9） and matrix metalloproteinase tissue inhibitor-1 （TIMP-1） in iliac artery of rabbits with atherosclerosis. Methods： Twenty male New Zealand white rabbits were studied. The rabbits were fed with a normal diet （normal control group, n=6） ; or a cholesterol diet （cholesterol. 1 g/d, and lard, 10 g/d for 6 weeks） and underwent balloon injury of iliac arteries two weeks after initiation of the diet （n= 16）, 8 of whom were pathological control group, 8 of whom were intravenous injected with puerarin （25 mg/kg · d per rabbit） after injury （puerarin group）. Four weeks after balloon injury, untreated and balloon injury iliac arteries were harvested for immunocytochemical staining. The mRNA and protein expression of MMP-9 and TIMP-1 were studied by RT PCR and immunucytochemistry. Results： The expression of MMP 9 mRNA （nRNA GAPDH mRNA） in normal control group, pathological control group and puerarin group was 0. 81±0. 17, 1.52±0. 24, 1.03±0. 19. respectively, with significant difference among them （P〈0.05）. The expression of TIMP-1 mRNA in above stated three-group was 1.44±0.14, 2.63±0. 16. 2.67±0. 12 respectively; the expression of TIMP-1 mRNA in pathological eontrol group and puerarin group were more than that of normal group （P〈0. 05）. but was no significant difference between pathological group and puerarin group （P〉 0. 05）. Puerarin inhibited MMP-9 protein expression （P〈0.05） and had no influence on TIMP-1 protein expression. Conclusion： Puerarin may make atheroseclerotic plaque stable by regulating of the expression of MMP-9.