葛根素对低O高CO肺动脉高压大鼠肺动脉平滑肌细胞线粒体途径凋亡的影响_论文.docx

葛根素对低O2高CO2肺动脉高压大鼠肺动脉平滑肌细胞线粒体途径凋亡的影响 【摘要】 目的: 研究在慢性低O2高CO2情况下，葛根素对肺动脉平滑肌细胞(PASMCs)线粒体途径凋亡的影响。方法:雄性SD大鼠40只，随机分为正常对照组(N组)、慢性低O2高CO2 2周组(H2w组)、慢性低O2高CO2 4周组(H4w组)、慢性低O2高CO2 4周＋葛根素组(H4w+G组)，每组10只。TUNEL法检测PASMCs凋亡；免疫组化测定PASMCs线粒体细胞色素C（Cyt-c）释放量及胞浆Bcl-2、Bax含量；分光光度法检测肺组织匀浆caspase 9酶活性。结果:①H2w组、H4w组和H4w+G组mPAP、RV/(LV+S)、PAMT、PASMCs胞浆 Bcl-2含量均显著高于N组（P ）。结论:葛根素有抑制慢性低O2高CO2肺动脉高压和肺血管重建的作用，促进PASMCs通过线粒体途径凋亡可能是其作用机制之一。 【关键词】 低氧 慢性 高血压 肺性 线粒体 凋 Abstract: Objective: To study the effects of puerarin on mitochondrial-dependent apoptosis pathway in pulomnary hypertension induced by hypoxic hypercapnia. Methods: Fourty Sprague-Dawley rats were randomly divided into four groups: normal control group(N)，hypoxic hypercapnic 2 weeks group (H2w)，hypoxic hypercapnic 4 weeks group (H4w) and hypoxic hypercapnic 4 weeks + Puerarin group (H4w+G). The apoptsis of PASMCs was tested with TUNEL. The release of Cyt-c from mitochondria and the content of Bcl-2 and Bax in cytoplasm were determined with immunohistochemistry. The activity of caspase 9 was detected with spectropho-tometric method. Results: ①The mPAP, RV/(LV+S), PAMT and the content of Bcl-2 of PASMCs of group H2w， group H4w and group H4w+G were significantly higher than those of group N (P ). Conclusion: Puerarin can lower pulmonary hypertension and inhibit structural remodeling of pulmonary arteries. Facilitating mitochondrial-dependent apoptosis of PASMCs may be one of the mechanisms of this action. Key words: hypoxia，chronic；hypertension，pulmonary；mitochondria；apoptosis；Puerarin 低氧性肺血管重建中各种类型细胞凋亡减少而增殖增加[1]。肺动脉平滑肌细胞(pulmonary ar-tery smooth muscle cells，PASMCs)是肺血管壁的主要组成部分，增加PASMCs凋亡可以逆转肺动脉高压肺血管重建[2]。细胞色素C(cytochrome C，Cyt-c) 从线粒体中释放是细胞线粒体途径凋亡的关键步骤[3,4]。线粒体Cyt-c的释放受到Bcl-2蛋白家族的调控，Bcl-2和Bax蛋白是该家族中具有代表性的蛋白，分别起到抑制和促进凋亡的作用[5,6]。葛根素(Puerarin)是豆科植物野葛干燥根中的主要有效成分,大量研究发现，葛根素能促进主动脉平滑肌细胞的凋亡[7]。本实验拟通过研究葛根素对PASMCs线粒体Cyt-c释放、caspase 9活性以及Bcl-2、Bax蛋白表达的影响，探讨葛根素促进PASMCs线粒体途径凋亡是否为其抑制肺动脉高压肺血管重建的作用机制。 1 材料和方法 材料 实验动物：雄性Sprague-Dawley（SD）大鼠40只，体重