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目的探讨半胱氨酸蛋白酶抑制剂C(cystatin C)对于自发性高血压大鼠(SHR)心肌纤维化的影响。方法培养16周龄的自发性高血压大鼠心肌成纤维细胞(CFs),将构建成功含cystatin C的腺病毒载体转染SHR的CFs及心室肌中,随机分为空白对照组、Ad-GFP组和Ad-CysC组。通过w estern blot检测cystatin C在CFs和SHR心室肌中的表达情况。用实时定量PCR测定CFs的Ⅰ、Ⅲ胶原mRNA水平。行超声心动检查评定转染后4周SHR的心脏结构和心功能。通过Masson染色分析SHR的心肌纤维化程度。结果 Cystatin C在Ad-CysC组CFs和心室肌中表达较空白对照组和Ad-GFP组明显增强(P均<0.05);与空白对照组和Ad-GFP组相比,Ad-CysC组CFs中Ⅰ、Ⅲ胶原mRNA表达显著增加(P均<0.05);Ad-CysC组LVMI、IVSD、LVPWD、CVF和PVCA明显高于空白对照组和Ad-GFP组,LVEDD较其余两组明显减小(P均<0.05)。上述指标在空白对照组和Ad-GFP组中差异均无统计学意义(P>0.05)。结论 Cystatin C使SHR成纤维细胞胶原合成增多,具有促进心肌纤维化进程的作用。
Objective To investigate the effect of cystatin C on myocardial fibrosis in spontaneously hypertensive rats (SHR). Methods 16-week-old spontaneously hypertensive rat cardiac fibroblasts (CFs) were cultured and transfected with cystatin C-containing adenovirus vector into CFs and ventricular muscle of SHR. The cells were randomly divided into blank control group, Ad-GFP Group and Ad-CysC group. The expression of cystatin C in ventricular myocytes of CFs and SHRs was detected by Western blot. Real-time quantitative PCR was used to detect the mRNA levels of collagen Ⅰ, Ⅲ in CFs. Echocardiography was performed to assess cardiac structure and cardiac function in SHR 4 weeks after transfection. The degree of myocardial fibrosis in SHR was analyzed by Masson staining. Results The expression of Cystatin C in CFs and ventricular myocytes in Ad-CysC group was significantly higher than that in blank control group and Ad-GFP group (all P <0.05). Compared with blank control group and Ad-GFP group, (P <0.05). LVMI, IVSD, LVPWD, CVF and PVCA in Ad-CysC group were significantly higher than those in blank control group and Ad-GFP group (P <0.05) All <0.05). There was no significant difference between the blank control group and Ad-GFP group (P> 0.05). Conclusion Cystatin C increases collagen synthesis in SHR fibroblasts and plays a role in promoting myocardial fibrosis.