不同浓度理气化痰活血方含药血清对大鼠心脏微血管内皮细胞活力的影响  

Effect of Different Concentration of LiQiHuaTanHuoXue Formula Containing Serum on Proliferation of Cardiac Microvascular Endothelial Cells in Rats

作　　者：葛昭 任思霖 周梦雪 李默涵 宁旭瑾 王贤良[1] GE Zhao;REN Si-lin;ZHOU Meng-xue;LI Mo-han;NING Xu-jin;WANG Xian-liang(The First Affiliated Hospital of Tianjin University of Traditional Chinese Medicine,Tianjin 300381,China;Graduate School of Tianjin University of Traditional Chinese Medicine,Tianjin 301617,China)

机构地区：[1]天津中医药大学第一附属医院,国家中医针灸临床医学研究中心,天津300381 [2]天津中医药大学研究生院,天津301617

出　　处：《时珍国医国药》2024年第12期2712-2718,共7页Lishizhen Medicine and Materia Medica Research

基　　金：国家自然科学基金面上项目(82174326);国家中医药管理局中医药创新团队及人才支持计划项目(ZYYCXTD-C-202203);国家教育部创新团队发展计划项目(IRT_16R54)。

摘　　要：目的探讨理气化痰活血方(LQHTHX)含药血清对缺氧/复氧诱导原代大鼠心脏微血管内皮细胞(CMECs)活力的影响。方法采用水煎煮法制备LQHTHX提取液并浓缩至高(2.7 g生药/mL)、中(1.35g生药/mL)、低(0.675 g生药/mL)3个剂量,按照10 mL/kg连续灌胃大鼠3d,在末次给药后0.5,1,1.5,2,2.5h采取血液样本分离血清,配置成5%、10%、15%以及20%的含药血清细胞培养基。提取新生大鼠原代CMECs并进行免疫荧光鉴定,体外构建细胞缺氧模型,分别加入不同浓度含药血清,采用CCK8法检测各组细胞活力,比较不同浓度含药血清处理后的细胞活力差异。结果成功提取了大鼠原代CMECs并构建了细胞缺氧模型。与模型组相比,阳性药物尼可地尔(200μmol/L)、缬沙坦(50μmol/L)以及中、高剂量组中不同浓度含药血清均能提高CMECs活力(P<0.01),其中以高剂量组中10%、15%LQHTHX含药血清组效果最好。结论LQHTHX可有效改善缺氧诱导的CMECs活力下降,从而发挥保护心脏微血管内皮功能的作用。Objective To investigate the effects of LiQiHuaTanHuoXue Formula(LQHTHX)on the viability of hypoxia/reoxygenation-induced primary cardiac microvascular endothelial cells(CMECs)in rats.Methods The LQHTHX extract was prepared by boiling in water and concentrated in three doses of high(2.7 g of raw material/mL),medium(1.35 g of raw material/mL),and low(0.675 g of raw material/mL),and then gavaged into rats for 3 days at 10 mL/kg.Blood samples were taken at 0.5 h,1 h,1.5 h,2 h,and 2.5 h after the last drug administration to isolate serum,which was prepared into 5%,10%,15%,and 20%drug-containing serum cell culture medium.The primary CMECs of neonatal rats were extracted and identified by immunofluorescence,and the cell hypoxia model was constructed in vitro.Different concentrations of drug-containing serum were added,and cell viability of each group was detected by the CCK8 method,comparing the differences in cell viability after treatment with different concentrations of drug-containing serum.Results In this study,we successfully extracted rat primary CMECs and constructed a cellular hypoxia model.Compared with the model group,the positive drugs,nicorandil(200μM)and valsartan(50μM),as well as different concentrations of drug-containing serums in the middle-and high-dose groups could increase the viability of CMECs(P<0.01),among which 10%and 15%LQHTHX-containing serums in the high-dose group had the best effect.Conclusion LQHTHX can effectively improve the hypoxia-induced decrease in the viability of CMECs,thus exerting a protective effect on the endothelial function of cardiac microvessels.This study established the foundation for further in-depth investigation of the pharmacological mechanism of the cardioprotective effect of LQHTHX.

关 键 词：理气化痰活血方 心脏微血管内皮细胞 细胞活力 冠状动脉微循环障碍 

分 类 号：R285.5[医药卫生—中药学]