半胱氨酸蛋白酶抑制剂C对自发性高血压大鼠心肌纤维化的影响

Effect of cystatin C on myocardial fibrosis in spontaneously hypertensive rats

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 Cardiac fibroblasts（CFs） of SHR aged 16 weeks were cultured in vitro.The recombinant adenovirus vector for cystatin C was transfected into CFs and the ventricular myocardium.All rats were randomly divided into 3 groups： the normal control group,the Ad-GFP group and the Ad-CysC group.Expression of cystatin C in CFs and the ventricular myocardium was determined by means of Western blot.MRNA expressions of collagen Ⅰ and Ⅲ were determined by RT-PCR.Echocardiography was used to evaluate the cardiac structure and function after transfection for 4 weeks.Myocardial fibrosis was analyzed with Masson′s staining.Results Protein expression of cystatin C was increased in CFs and the ventricle myocardium in the Ad-CysC group when compared with the normal control group and the Ad-GFP group（both P0.05）.Compared with the normal control group and the Ad-GFP group,mRNA expressions of collagenⅠ and Ⅲ were increased in the Ad-CysC group（both P0.05）;The LVMI,IVSD,LVPWD,CVF and PVCA were more increased and the LVEDD was more decreased in the Ad-CysC group than the normal control group and the Ad-GFP group（all P0.05）,while there was no statistical significance between the Ad-GFP group and the normal control group（P0.05）.Conclusion Cystatin C promotes collagen synthesis in CFs and may play a role in the progression of myocardial fibrosis in SHR.