摘要
目的探究甘西鼠尾草对培养H9c2大鼠心肌细胞缺氧/复氧损伤的保护作用及机制。方法采用体外细胞培养方法培养乳鼠心肌细胞,建立缺氧/复氧损伤心肌细胞模型,用甘西鼠尾草进行细胞给药,干预细胞生长过程。将培养的H9c2大鼠心肌细胞随机分为6组,分别为缺氧/复氧模型组(hypoxia/reoxygenation,H/R组);正常对照组(control,C组);缺氧/复氧模型+维拉帕米(verapamil)阳性对照组(H/R+V组);缺氧/复氧模型+甘西鼠尾草低、中和高剂量(0.01、0.1和1.0mg·L^(-1))干预组(H/R+L、M、H组)。使用噻唑蓝(thiazolyl blue tetrazolium,MTT)法测定细胞存活率;收集所培养的细胞上清液测定丙二醛(malondialdehyde,MDA)的含量和乳酸脱氢酶(lactate dehydrogenase,LDH)的活性;使用荧光酶标仪,测定能够反映细胞内活性氧(reactive oxygen species,ROS)水平高低的荧光吸光度值。结果甘西鼠尾草可明显提高心肌细胞存活率,有效减少细胞内LDH漏出量和胞浆内MDA的含量,并降低细胞内ROS水平。结论甘西鼠尾草对缺氧/复氧损伤心肌细胞具有显著的保护作用,其作用机制可能与其增强细胞清除氧自由基能力、减少脂质过氧化物的产生有关。
Objective To study the protective effects and the mechanism of Salvia przewalskii on cultured H9c2 rat cardiomyocytes treated by hypoxia/reoxygenation. Methods the hypoxia/reoxygen （H/R） injury cardiomyocytes model was established in H9c2 cell strain with or without Salvia przewalskii treated. Cultured H9c2 rat cardiomyocytes were randomly divided into six groups ： control group （ C group ） ; H/R group ; H/R ＋ Verapamil （ H/R ＋ V 组） group ; H/R ＋ Salvia przewalskii low concentration group （ 0.01 mg·L^-1 ） , H/R ＋ Salvia przewalskii middle concentration group（0. 1 mg·L^-1 ）, H/R ＋ Salvia przewalskii high concentration group （ 1.0 mg·L^-1 ）. Cell survival rate was determined using the 3- （4,5-dimethylthiazol-2-yl） -2,5- diphenyltetrazolium bromide （MTT） assay. The activity of lactate dehydrogenase （LDH） and the content of Malondialdehyde （MDA）was measured respectively in cellular supemate. The fluorescence absorbance values reflecting the level of intracellular reactive oxygen species （ ROS ） was measured by using Fluorescence microplate reader. Results The groups being treated with Salvia przewalskii could effectively increase the cell survival rate and decrease the release of LDH. Moreover, they could decrease the production of MDA and the level of ROS. Conclusions It is suggested that Salvia przewalskii has a protective effect on H9c2 rat cardiomyocyte treated by H/R injury. The mechanisms of the protective effects of Salvia przewalskii are related to enhance the capability of the cell clearing the oxygen free radical and to decrease the production of lipid peroxidation.
出处
《沈阳药科大学学报》
CAS
CSCD
北大核心
2016年第2期140-143,164,共5页
Journal of Shenyang Pharmaceutical University
基金
深圳市基础研究项目-中药丹参质量的化学模式识别研究资助项目(JYCJ20130402144215891)
