摘要
目的:观察痰瘀同治方含药血清对氧化型低密度脂蛋白(ox-LDL)所致人脐静脉内皮细胞(HUVECs)损伤的保护作用及探讨其抗动脉粥样硬化(AS)的作用机制。方法:体外培养HUVECs,分别以痰瘀同治方和辛伐他汀含药血清预处理细胞2 h,然后加入100 mg.L-1ox-LDL作用24 h。MTT法检测细胞活力;Griess法检测细胞上清中一氧化氮(NO)含量变化;Real-time RCR法检测小窝蛋白-1(Cav-1)和内皮型一氧化氮合酶(eNOS)mRNA表达;Western blott检测Cav-1和eNOS蛋白表达。结果:HUVECs经100 mg.L-1ox-LDL刺激后细胞活力显著降低(P<0.01);加入不同剂量痰瘀同治方和辛伐他汀含药血清后,细胞活力显著升高,细胞上清中NO含量明显提高(P<0.05)。此外,痰瘀同治方和辛伐他汀含药血清可下调Cav-1mRNA和蛋白表达和上调eNOS mRNA和蛋白表达,其中以辛伐他汀和痰瘀同治方高剂量含药血清作用尤为显著(P<0.01)。结论:痰瘀同治方能够通过提高NO含量,下调Cav-1表达和上调eNOS表达起到内皮细胞保护作用,可能是其抗AS分子机制之一。
Objective： To observe the protective effect of serum containing Tanyu Tongzhi formula（TYTZF） on ox-LDL-induced human umbilical vein endothelial cells（HUVECs） injury and detect its mechanism on anti-artherosclerosis.Method： HUVECs were pretreated with serum containing TYTZF and simvastatin respectively for 2 hours and then added with 100 mg·L-1 of ox-LDL and laid aside for 24 hours.The activity of HUVECs were measured by the methyl thiazolyl tetrazolium（MTT） method and the NO content in cell culture supernatants were examined by the Griess method.The mRNA levels of Cav-1 and eNOS were measured by the Real-time PCR method.The protein expression of Cav-1 and eNOS were detected by Western blotting.Result： The activity of HUVECs was significantly decreased after ox-LDL treatment（P0.01） and this decrease was significantly inhibited by serum containing TYTZF and simvastatin of different doses（P0.05）.They could enhance the NO content in cell culture supernatants,down-regulate the expression of Cav-1 and up-regulate the expression of eNOS at mRNA and protein levels,which was especially notable after treatment with serum containing TYTZF and simvastatin in large doses.Conclusion： TYTZF has the protective effect on HUVECs by increasing the production of NO and up-regulating Cav-1 expression and down-regulating eNOS expression.
出处
《中国中药杂志》
CAS
CSCD
北大核心
2012年第7期974-978,共5页
China Journal of Chinese Materia Medica
基金
国家"重大新药创制"科技重大专项(2009ZX09502-017
2009ZX09301-005
2009ZX09301-005-2-6)
