摘要
目的:以肠吸收液为药物载体,利用拆方重组法筛选脑心通胶囊组方中发挥体外心肌保护作用的单味或多味中药,并丰富脑心通胶囊心肌保护作用的分子机制。方法:应用外翻肠囊法制备肠吸收液,体外构建过氧化氢(H2O2)损伤大鼠胚胎心肌细胞株(H9c2)模型,分为正常组、模型组、脑心通胶囊肠吸收液低、中、高剂量组(15.63,31.25,62.50 mg·L-1),每组3个复孔,实验重复3次。噻唑蓝(MTT)比色法检测心肌细胞的存活率,AnnexinⅤ-FITC/PI双染结合流式细胞仪检测心肌细胞凋亡率,蛋白免疫印迹法(Western blot)检测水通道蛋白1(AQP1)蛋白的表达;同时拆方或构建重组方,MTT比色法定位脑心通胶囊组方中发挥心肌保护作用的单味或多味中药。结果:与正常组比较,模型组AQP1的表达升高(P<0.01),心肌细胞凋亡数量增多(P<0.05)。与模型组比较,脑心通胶囊肠吸收液中、高剂量组均可下调AQP1的表达(P<0.05),脑心通胶囊肠吸收液高剂量组可抑制心肌细胞凋亡(P<0.05)。与脑心通全方组比较,丹参、地龙和全蝎这3种单味药心肌保护作用与脑心通全方组相当,此3味药重组方高剂量组心肌保护作用强于脑心通全方组(P<0.05),去3味药重组方心肌保护作用弱于脑心通全方组(P<0.05)。结论:脑心通胶囊通过抑制AQP1表达和细胞凋亡,发挥心肌保护作用;丹参、地龙和全蝎是组方中发挥心肌保护作用的主要药味。
Objective: Intestinal absorption liquid as medicine carriers,one or more kinds of traditional Chinese medicine protecting cardiac injury in Naoxintong capsule will be found through separation and combination of prescription. This study also can expand protecting mechanisms of Naoxintong capsule. Method: Naoxintong intestinal absorption liquids of single,combination and total prescription were prepared. H9c2 cell line exposed to H2O2 was established. Cell survival rate was determined with methyl thiazolyl tetrazolium ( MTT) . Cell apoptosis rate was examined by Annexin Ⅴ-FITC /PI staining with a flow cytometer. Aquaporin1 ( AQP1) expression was detected by Western blot. One or more kinds of traditional Chinese medicine in Naoxintong capsule which exerted protective effect from cardiac injury were screened through separation and combination of prescription. Result: As compared with control group,the protein expression level of AQP1 was significantly increased ( P < 0. 01) ,and the apoptotic H9c2 cells increased ( P < 0. 05) . Compared with model group,Naoxintong intestinal absorption liquid low,medium and high dose group decreased AQP1 expression ( P < 0. 05 ) ,and naoxintong intestinal absorption liquid high dose group inhibited cardiac cells apoptosis ( P < 0. 05) ,thus protected H9c2 cells from oxidative damages. Salviae Miltiorrhizae Radix et Rhizoma,Pheretima,Scorpio are main constituents which had protective actions in total prescription. Protective effect of those three combination prescription in high dose group was the best ( P < 0. 05) ,and effect of combination prescription without those three was the second ( P < 0. 05) . Conclusion: Naoxintong capsule acts as a protective role in myocardial injury through decreasing AQP1 expression and inhibiting cell apoptosis. Salviae Miltiorrhizae Radix et Rhizoma,Pheretima,Scorpio are important components in Naoxintong capsule.
作者
何子龙
赵晔
张毅
李德凤
张方博
杨洪军
HE Zi-long;ZHAO Ye;ZHANG Yi;LI De-feng;ZHANG Fang-bo;YANG Hong-jun(Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beijing 100700,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2019年第4期29-35,共7页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家"重大新药创制"科技重大专项(2011ZX09201-201-26)
国家自然科学基金青年基金项目(81403207)
关键词
脑心通胶囊
肠吸收液
心肌损伤
过氧化氢
有效药味筛选
Naoxintong capsule
intestinal absorption liquid
cardiac injury
hydrogen peroxide
active medicinal identification
