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蕨麻乙醇提取物抑制大鼠急性心肌缺血再灌注损伤导致的心肌细胞凋亡

Keywords: 蕨麻,心肌缺血再灌注,细胞凋亡,TUNEL,Caspase

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Abstract:

目的:观察蕨麻乙醇提取物对大鼠结扎冠状动脉所致急性心肌缺血再灌注损伤导致的心肌细胞凋亡的保护作用,以及对心肌细胞凋亡信号通路中天冬氨酸特异性半胱氨酸蛋白酶9、天冬氨酸特异性半胱氨酸蛋白酶3(Caspase-9,Caspase-3)表达的影响。方法:选取雄性SD大鼠96只随机分为手术对照组,模型组,蕨麻乙醇提取物0.9,1.8,3.6g·kg-1干预组,阳性药物恬尔心组(盐酸地尔硫卓,diltiazem,30mg·kg-1)。通过结扎大鼠冠状动脉左前降支建立急性心肌缺血再灌注模型。采用原位末端标记法检测心肌细胞凋亡;RT-PCR检测心肌组织Caspase-3,Caspase-9mRNA水平;免疫组化染色半定量检测Caspase-3,Caspase-9蛋白水平。结果:TUNEL结果提示心肌缺血再灌注损伤后,心肌细胞出现大量凋亡,模型组凋亡指数为(31.5±3.6)%;各组蕨麻乙醇提取物均明显抑制缺血再灌注损伤所致的心肌细胞凋亡。模型组细胞凋亡导致Caspase-3,Caspase-9mRNA及蛋白大量表达,而给予蕨麻乙醇提取物后,1.8,3.6g·kg-1剂量组缺氧损伤心肌Caspase-9mRNA水平显著降低(P<0.05),0.9g·kg-1剂量组与模型组相比未见明显差异。各剂量组蕨麻乙醇提取物均可显著降低心肌组织Caspase-3mRNA水平(P<0.05)。免疫组化分析提示各干预组蕨麻乙醇提取物均可显著抑制Caspase-3,Caspase-9蛋白的表达(P<0.05)。结论:蕨麻乙醇提取物能够显著抑制急性心肌缺血再灌注所致心肌细胞凋亡,并抑制Caspase-3,Caspase-9mRNA水平及蛋白水平的表达。

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