欣力胶囊抑制Angiotensin-2刺激引起的心肌细胞损伤

Xinli Capsule prevents cardiomyocytes from injury induced by angiotensin Ⅱ

目的在原代培养新生大鼠心肌细胞水平,观察欣力胶囊对血管紧张素Ⅱ(AngⅡ)刺激引起的心肌细胞损伤的保护作用。方法采用免疫组织化学方法(α-actin)鉴定原代心肌细胞纯度。以欣力胶囊灌胃大鼠进行血清药理学实验;将心肌细胞分组为:正常对照组、AngⅡ刺激组和欣力胶囊含药血清治疗组(包括3个不同剂量组)。倒置显微镜下于不同时段计数心肌细胞搏动频率;用反转录聚合酶链反应(RT-PCR)检测心肌细胞肥大早期基因(c-myc、c-jun和c-fos)的表达改变;利用SABA—18生化分析仪测定不同分组细胞培养液上清乳酸脱氢酶(LDH)、肌酸激酶(CK-MB)以及天冬氨酸转氨酶(GOT)含量。结果通过α-actin免疫组织化学染色鉴定,心肌细胞纯度达95%,满足实验需要。欣力胶囊含药血清在体外能够有效抑制AngⅡ刺激引起的搏动频率增加(P<0.05);抑制心肌细胞肥大早期基因(c-myc、c-jun和c-fos)的表达;减少细胞培养液上清中心肌酶(LDH、CK-MB、GOT)的含量(P<0.05)。结论欣力胶囊可以抑制AngⅡ刺激对原代培养大鼠心肌细胞的损伤。

Objective To investigate the treatment effects of Xinli capsule （XL） on Angiotensin Ⅱ-induced injury in primary＇ cuhure of neonatal rat eardiomyoeytes. Methods The ratio of cultured cardiac myoeytes obtained from neonatal hearts were calculated by immunohistoehemieal expression of α-aetin.To investigate the effects of XL in vitro, the method of serum pharmacology was introduced, Cultured cells were divided into three groups： ① normal controls, ②cells stimulated by angiotensin- Ⅱ （1 μmol/L）, and ② cells treated with XL and angiotensin- Ⅱ. In third group, there are three subgroups according to the variety of dosage. Then the pulsation frequency of cardiac myoeytes were, calculated, and the expression of early response genes, e.g. c-myc, c-jun and c-los, were determined by reverse transcription polymerase chain reaction （RT-PCR）. In addition, the concentrations of myocardial enzymes were obtained by biochemistry analysis. Results The ratio of cardiac myoeytes reached 95% and met the requirements of subsequent experiment. Treatment with Ang Ⅱ increased neonatal rat cardiaomyoeyte pulsation frequency, expression of early response genes, and concentrations of myocardial enzymes （vs, normal control, P〈0.01）. Co-treatment with serum from rats treated with XL inhibited the Ang Ⅱ -induced injury （vs. Ang Ⅱ alone, all P〈0.01）. Conclusion These findings indicated that Xinli Capsule may attenuate cardiomyocyte injury induced by Angiotensin Ⅱ.