九龙藤总黄酮对缺氧/复氧心肌损伤的作用

Effects of Bauhinia championii Flavones on Hypoxia-Reoxygenation Injury in Myocardial Cells

目的研究九龙藤总黄酮对缺氧/复氧乳鼠心肌细胞的影响及作用机制。方法建立心肌细胞缺氧/复氧损伤模型,给予不同剂量九龙藤总黄酮(终质量浓度分别50、100、200 mg·L-1)预处理,倒置显微镜下观察心肌细胞形态学变化;酶联免疫法测定肿瘤坏死因子活性;免疫组化方法观察Bcl-2、Bax、NF-κB蛋白的表达;Annex v-FITC/PI双染法测定心肌细胞凋亡率。结果与模型组相比,九龙藤总黄酮预处理能减轻心肌细胞损伤,降低肿瘤坏死因子-α、诱导型一氧化氮合酶、维持内皮型一氧化氮合酶水平,上调Bcl-2、下调Bax、NF-κβ蛋白表达,抑制心肌细胞凋亡(P<0.01或P<0.05)。结论九龙藤总黄酮对缺氧/复氧心肌损伤具有保护作用,其机制可能与调节诱导型一氧化氮合酶、核转录因子-κβ信号通路,上调Bcl-2、下调Bax和核转录因子-κB蛋白表达,抑制心肌细胞凋亡有关。

OBJECTIVE To observe the effect and mechanisms of Bauhinia championii flavones （BCF） on hypoxia/reoxygenation （H/R） myocardial ceils of neoanatal rats. METHODS The cardiomyocytes hypoxia/reoxygenation injury model was developed, and Bauhinia championii flavones was pretreated （final concentration as 50, 100, 200 mg·L-1 respectively）. The morphological changes of cardiomyocytcs were observed by invert microscope. ELISA was used to evaluate the activities of tumor necrosis factor alpha, the protein expression of Bcl-2, Bax and NF-κB was observed by immunohisto-chemistry, and Annex v-FITC/PI staining was used to detect apoptosis rate. RESULTS Compared with model group, Bauhinia championii flavones pretreatment eased cardiomyocytes injury, decreased the activities of TNF-a and iNOS, maintained the eNOS level, up-regulated the expression of Bcl-2, down-regulated Bax and NF-κB, and inhibited cadiocyte apoptosis（P 〈 0. 01 or P 〈 0. 05）. CONCLUSION BCF has protective effects on hypoxia/reoxygen- ation injury, which are associated with adjusting iNOS and NF-κB signal channel, up-regulating Bcl-2, down-regulating Bax and NF- κB, and inhibiting cadiocyte apoptosis.