论文部分内容阅读
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 anti-hypertrophy effect of Huatan Huoxi Recipe serum and its effect on type I and type III collagen. Methods: Saline, Phlegm Huoxue Yiqi low, medium and high dose (24.42,48.83,97.66 g / kg) and captopril (0.1 g / kg) serum were prepared. Rat H9c2 cardiomyocyte hypertrophy induced by isoproterenol (ISO) was used (10μmol / L, 48 h). Immuno-fluorescent staining and cell surface area determination were performed after intervention of 20% drug-containing serum in each group. Cell protein concentration was measured by BCA method. Atrial natriuretie peptide (ANP) gene was detected by real-time fluorescent quantitative RT-PCR. The expression of brain natriuretic peptide (BNP) and type I and type III collagen mRNA was detected by Western blot. Results: Compared with the normal control group, the expression of mRNA and protein of ANP and BNP in model group were significantly increased (P <0.05). Compared with the model group, the above indexes of Huatan Huoxue Yiqi medium dose group were significantly lower (P <0.05). Conclusion: (1) Huatan Huoxue Qi medicated serum can effectively improve ISO-induced H9c2 cardiomyocyte hypertrophy and reduce the expression of type I and type III collagen; (2) In the group of cells, the medium dose group which is equivalent to the adult dose is the most obvious improvement, and the effect is better than captopril group.