摘要
目的:研究当归红芪超滤膜提取物(UFE-AH)对阿霉素所致心肌细胞凋亡的影响。方法:取新生1~2 d的Wistar乳鼠,用差速贴壁法原代提取心肌细胞,调整细胞密度1×106/mL,于37℃5%CO2培养箱中密闭培养48 h。用2.5 mg.L-1阿霉素作用于心肌细胞24 h成功建立心肌细胞凋亡模型。实验分5组:正常对照组、阿霉素组(2.5 mg.L-1)、UFE-AH低、中、高剂量干预组(阿霉素造模前2 h预先分别给予3.75,7.5,15 g.L-1当归红芪超滤膜提取物)。四氮唑蓝(MTT)法检测各组细胞生长抑制率,单细胞凝胶电泳检测DNA损伤,Western blot(蛋白印迹法)检测B淋巴细胞瘤-2蛋白(Bcl-2),Bcl-2相关X蛋白(Bax)表达水平。结果:与正常组相比,阿霉素组心肌细胞生长抑制率明显增加,且头部DNA含量占总含量的百分数(HDNA%),尾部DNA含量占总含量的百分数(TDNA%),尾长(TL),尾短(TM),尾距(OTM)均明显升高,Bcl-2蛋白表达水平下降、Bax表达水平升高,均有统计学意义(P<0.01);UFE-AH干预组的心肌细胞生长抑制率明显降低,且HDNA%,TDNA%,TL,TM,OTM数值均降低,Bcl-2蛋白表达水平上调,Bax表达水平下调,有显著性差异(P<0.01)。结论:UFE-AH可减轻阿霉素对心肌细胞DNA的损伤,上调Bcl-2/Bax而抑制心肌细胞的凋亡,具有抗心肌细胞凋亡的保护作用。
Objective: To investi gate the influence of ultra-filtration extract from the mixture of Angelica sinensis and Hedysarum polybotrys(UFE-AH) on myocardial cell apoptosis induced by adriamycin(ADR) of neonatal rats.Method: Myocardial cells of neonatal rats born 1 to 2 days were primarily cultured by differential adherent method for 48 h at 37 ℃,5% CO2 incubator,adjusting the cell concentration to 1 × 106/ mL,then randomly divided into five group: normal control group,adriamycin group,low-does,medium-does,and highdoes UFE-AH.to detect MTT method was used to inhibition of cell growth rate,and to detect the level of deoxyribonucleic acid damage(DNA damage) the single cell gel electrophoresis was used,western blot was used to detect the expression of Bcl-2 and Bax.Result: Compared with the normal group,the inhibition of cell growth rate in adriamycin group increased significantly,head DNA%(HDNA%),tail DNA%(TDNA%),tail length(TL),tail moment(TM),olive tail moment(OTM) were significantly increased,the protein expressions of Bcl2 were decreased and the protein expressions of Bax were increased significantly(P〈0.01);In comparison of the four other groups with model group,UFE-AH,HDNA%,TDNA%,TL,TM,OTM numerical and the protein expressions of Bax,but increase the protein expressions of Bcl-2 of the myocardial ceels in neonatal rats and the radio of Bcl-2 / Bax.Conclusion: can reduce the DNA damage of the myocardial cells induced by adriamycin and increase the radio of Bcl-2 / Bax,and has the protective effects on myocardial cell apoptosis induced by adriamycin of neonatal rats.
出处
《中国实验方剂学杂志》
CAS
北大核心
2013年第15期222-226,共5页
Chinese Journal of Experimental Traditional Medical Formulae
基金
甘肃省中医药科研课题(07-gzk-31)