益肺活血颗粒对缺氧大鼠肺动脉平滑肌细胞内低氧诱导因子-1α和活性氧的影响  

Effect of Yifeihuoxue granule on the hypoxia inducible factor-1 alpha and reactive oxygen species of rat pulmonary artery smooth muscle cells cultured in hypoxia condition in vitro

作　　者：张凌云[1] 欧敏[2] 顾珏[2] 宋秀杰[2] 

机构地区：[1]三峡大学第一临床医学院-宜昌市中心人民医院呼吸内科,湖北宜昌443003 [2]海军总医院干部呼吸内科,北京100048

出　　处：《转化医学杂志》2012年第2期69-72,共4页Translational Medicine Journal

基　　金：全军医药卫生科研基金资助项目(36040)

摘　　要：目的观察益肺活血颗粒对低氧培养大鼠肺动脉平滑肌细胞(pulmonary artery smooth musclecells,PASMCs)内低氧诱导因子-1α(hypoxia inducible factor-1 alpha,HIF-1α)和活性氧(reactive oxygen species,ROS)的影响。方法采用血清药理学方法制备不同浓度的益肺活血颗粒(yifeihuoxue granule,YFHXG)含药血清,采用组织块贴壁法原代培养大鼠PASMCs,取对数生长期PASMCs随机分为常氧组、缺氧组、缺氧+YFHXG组(16.5、3.3、0.66 g/kg)。用噻唑蓝比色法测定各组PASMCs的增殖效应,免疫组化法测定细胞内HIF-1α蛋白的表达,激光共聚焦显微镜测定细胞内ROS的含量。结果与常氧组相比,缺氧组PASMCs增殖明显活跃,HIF-1α蛋白表达及ROS含量增加;与缺氧组相比,缺氧+YFHXG高、中浓度组大鼠PASMCs的生长明显受抑制,而且HIF-1α蛋白表达及ROS含量降低。结论缺氧可以直接刺激PASMCs的增殖。YFHXG可以显著抑制低氧对大鼠PASMCs的促增殖作用,其机制可能是通过降低细胞内HIF-1α蛋白表达及ROS的水平来实现的。Objective To investigate the effect of Yifeihuoxue granule （YFHXG） on the hypoxia inducible factor-1 alpha （ HIF-1α ） and reactive oxygen species （ ROS ） of rat pulmonary artery smooth muscle cells（PASMCs） exposed to hypoxic conditions. Methods Serum-pharmacology methods were used in the preparation of YFHXG serum, tissue block anchorage were performed in the primary culture of rat PASMCs. PASMCs in exponential phase were randomly divided into normoxia group,hypoxiagroup, hypoxia ＋YFHXG group （ 16.5,3.5 g/kg and 0.66 g/kg ）. Cell viability was assessed with 3- （ 4,5-Dimethyhhiazol-2-yl ） -2,5-diphenyhetrazolium bromide （ MTT） assay. In addi- tion, HIF-1α protein expression was evaluated by immunocytochemistry analysis, the concentration of intracellular ROS was determined by laser scanning confocal microscopy（LSCM）. Results MTI＂ assay showed that hypoxia could directly activate the proliferation of PASMCs, while YFHXG dose- dependently inhibited hypoxia-induced proliferation of rat PASMCs. Immunocytochemistry showed that hypoxia enhanced HIF-Iot protein expression, and LSCM showed that hypoxia significantly increased intracellular ROS, while YFHXG decreased the expression of HIF-1α and attenuated the hypoxia-induced increase in intracellular concentration of ROS. Conclusion YFHXG can inhibit hypoxia-induced proliferation of rat PASMCs. The anti-hypoxia effect of YFHXG may be explained by its regulation of HIF-1α expression and the levels of intracellular ROS.

关 键 词：益肺活血颗粒 肺动脉平滑肌细胞 低氧 低氧诱导因子-1Α 活性氧 

分 类 号：R916.3[医药卫生—药学] R322.121