Abstract:

Objective    To explore the  effects of cystatin（CysC）on the proliferation of rat cardiac fibroblasts. Methods    Cardiac fibroblasts from 1～3 days old Sprague-Dawley rats (clean grade, weight 10～15g) were cultured by the anchorage velocity-dependent separation method. The recombinant adenovirus vector for CysC gene（Ad-CysC）was constructed with the GatewayTM technique and transfected into rat cardiac fibroblasts in vitro. Rat cardiac fibroblasts were randomly divided into the normal control group, the Ad-GFP group and the Ad-CysC group. Expression of CysC at the protein level was examined by means of Western-blot .Cell proliferation rate was measured by bromodeoxyuridine（BrdU）. Results    DNA sequencing results and Western blot analysis indicated that recombinant Ad-CysC was successfully constructed. The efficiency of Ad-CysC infecting rat cardiac fibroblasts was above 90% when multiplicity of infection(MOI) was 200. Furthermore,  protein expression of CysC was significantly increased at 24h after transfection（P<0.05）. Compared to the normal control group and the Ad-GFP group, the BrdU incorporation rate of the Ad-CysC group was much higher at 72h after transfection（P<0.05）. Conclusions    High expression of CysC can stimulate the proliferation of cardiac fibroblasts，which could promote the myocardial fibrosis.

Key words: CystatinC；Cardiac fibroblast；Cell proliferation