化痰活血益气方含药血清对异丙肾上腺素作用下大鼠H9c2细胞株Ⅰ、Ⅲ型胶原的影响  

Effects of Drug Serum of Huatan Huoxue Yiqi Formula on Collagen Ⅰ and Collagen Ⅲin H9c2 Cells with Isoproterenol-Induced Cardiac Hypertrophy

作　　者：尹宝[1] 曹思[1,2] 张林[1] 陈涛[2] 韩立民[2] 

机构地区：[1]湖南中医药大学,湖南长沙410208 [2]赣南医学院,江西赣州341000

出　　处：《中药材》2017年第4期931-936,共6页Journal of Chinese Medicinal Materials

基　　金：湖南中医药大学博士研究生创新课题项目(2016CX07)

摘　　要：目的:探讨化痰活血益气方含药血清的抗心肌肥厚作用及对细胞间质Ⅰ、Ⅲ型胶原的影响。方法:分别制备生理盐水、化痰活血益气方低、中、高剂量(24.42、48.83、97.66 g/kg)及卡托普利(0.1 g/kg)含药血清。采用异丙肾上腺素(Isoproterenol,ISO)诱导的大鼠H9c2心肌细胞肥厚模型(10μmol/L,48 h)。各组细胞给予相应20%含药血清干预后,行免疫荧光染色并测定细胞表面积,BCA法检测细胞蛋白浓度,实时荧光定量RT-PCR检测胚胎基因心房利钠肽(atrial natriuretie peptide,ANP)、脑钠肽(brain natriuretic peptide,BNP)及Ⅰ、Ⅲ型胶原mRNA的表达,Western blot检测Ⅰ、Ⅲ型胶原蛋白的表达。结果:与正常对照组比较,模型组细胞的表面积、蛋白浓度、胚胎基因(ANP、BNP)mRNA及Ⅰ、Ⅲ型胶原mRNA和蛋白表达均显著升高(P<0.05)。与模型组比较,化痰活血益气方中剂量组上述指标均显著降低(P<0.05)。结论:(1)化痰活血益气方含药血清可有效改善ISO诱导的H9c2心肌细胞肥厚并降低Ⅰ、Ⅲ型胶原的表达;(2)在化痰活血益气方含药血清干预的各组细胞中,等效于成人给药剂量的中剂量组细胞改善最为显著,且效果优于卡托普利组。Objective：To investigate the effects of drug serum of Huatan Huoxue Yiqi formula on collagen Ⅰ and collagen m in H9c2 cells with isoproterenol （ ISO ） -induced cardiac hypertrophy. Methods ： Drug serum of normal saline, different doses of Huatan Huoxue Yiqi formula（24. 42,48.83,97.66 g/kg） and captopril（0. 1 g/kg） were prepared,respectively. 10 μmol/L ISO was used to stimulate cardiomyocyte cell hypertrophy model for 48 h, and respective 20% drug serum was given to the cells. Cell surface area was measured by immunofluorescence method. Cell protein concentration was detected by BCA method, mRNA levels of atrial natriuretie peptide （ANP）, brain natriuretic peptide（ BNP）, collagen Ⅰ and collagen Ⅲ were detected by Real-time RT-PCR, respectively. Protein expressions of collagen Ⅰ and collagen m were detected by Western blot. Results ： Compared with control group,the cell surface area, cell protein concentration, mRNA levels of mRNA（ ANP ,BNP）, collagen Ⅰ and collagen Ⅲ in mRNA and protein expressions were all increased significantly（P 〈0. 05）. Compared with model group, all the indexes above were significantly decreased at different dose in Huatan Huoxue Yiqi formula groups and Captopril group （P 〈 0. 05 ）. Conclusion ：（1）Drug serum of Huatan Huoxue Yiqi formula can attenuate ISO-induced cardiac hypertrophy and decrease the expression of collagen Ⅰ and collagen Ⅲ in vitro. （2）In Huatan Huoxue Yiqi formula groups, the effect of formula middle dose equivalent to human clinical dose is the best and which is better than captopril.

关 键 词：化痰活血益气方 含药血清 H9C2 心肌肥厚 Ⅰ型胶原 Ⅲ型胶原 

分 类 号：R285.5[医药卫生—中药学]