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    • TFPI对大鼠心肌缺血再灌注损伤的影响及其机制的初步探索

    李嘉舒, 高薇, 代悦, 徐睿, 沈丽,闫汝楠, 刘越,陈文佳,刘文秀,傅羽

    cnki（2023）

    Cited 0|Views9
    Abstract
    目的 探究组织因子途径抑制物（TFPI）对大鼠心肌缺血再灌注（I/R）及心肌细胞缺氧复氧（H/R）损伤的影响，并从心肌细胞凋亡的变化探索其机制。方法 在体内实验中，通过SD大鼠心脏原位结扎法可逆阻断前降支建立大鼠心肌I/R模型。将大鼠随机分为对照组、I/R组和I/R+rTFPI组，再灌注后3天采用HE染色观察大鼠心肌组织形态学变化，TTC染色法评估心肌梗死区范围，扫描透射电镜观察心肌超微结构损伤情况，Western-Blot法检测各组大鼠心肌组织中Bcl-2、Bax和cleaved-caspase-3蛋白的表达。在体外实验中，采用胰酶消化法及差速贴壁法培养SD乳鼠原代心肌细胞，用MIC101系统模拟心肌细胞I/R损伤，缺氧2小时、复氧12小时后建立体外心肌细胞缺氧/复氧（H/R）模型。将心肌细胞分为对照组、H/R组和H/R+rTFPI（10μg/L）组，用CCK8法检测心肌细胞活力，TUNEL法检测心肌细胞凋亡率，Western-blot方法检测心肌细胞中Bax、Bcl-2及cleaved-caspase-3蛋白的表达水平。结果 体内实验中，成功建立大鼠在体心肌I/R模型。HE染色结果显示I/R组较对照组心肌细胞坏死程度加重，I/R+rTFPI组较I/R组心肌细胞坏死程度减低；TTC染色示I/R+rTFPI组较I/R组心肌梗死范围减少了39.76%（P＜0.05）；扫描透射电镜观察显示I/R组凋亡及损伤程度较对照组加重，I/R+rTFPI组凋亡及损伤较I/R组减轻；Western-Blot结果示，再灌注3天后I/R组心肌组织Bcl-2的表达较对照组降低了53.43%（P＜0.05）、Bax和cleaved-caspase-3（P＜0.05）的表达较对照组分别增加了29.05%和73.25%（P＜0.05），而I/R+rTFPI组Bcl-2的表达水平较I/R组升高了55.01%（P＜0.05），Bax和cleaved-caspase-3的表达水平较I/R组分别降低了13.77%和24.25%（P＜0.05）。在体外实验中，CCK8检测结果显示H/R组细胞活力较对照组下降了29.70%（P＜0.05），H/R+rTFPI组细胞活力较H/R组升高了19.77%（P＜0.05）。TUNEL结果显示H/R组较对照组凋亡率增加了56.76%，H/R+rTFPI组细胞凋亡率较H/R组降低了24.55%（P＜0.05）。Western-blot结果示：H/R组细胞Bcl-2表达较对照组降低了46.92%，Bax表达较对照组增加了41.90%（P＜0.05），cleaved-caspase-3表达较对照组升高了2.68倍（P＜0.05）。H/R+rTFPI组Bcl-2表达较H/R组增加了28.24%（P＜0.05），Bax及cleaved-caspase-3表达较H/R组分别降低了26.34%和57.60%（P＜0.05）。结论 TFPI可显著拮抗心肌I/R和心肌细胞H/R损伤，此效应与其抑制心肌细胞凋亡有关。
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