摘要
目的研究胱抑素C(CysC)对人动脉平滑肌细胞(ASMC)中组织蛋白酶S(CatS)和细胞外基质的影响。方法以正常ASMC为空白对照组,RT-PCR及Western-blot法测定CysC高表达质粒转染(CysC高表达组)及小干扰RNA(siRNA)分别干扰CysC(干扰组)0、12、24、48、72h后CatS的表达;ELISA法测定空白对照组、CysC高表达24h组和48h组、干扰48h组和72h组中Ⅰ型胶原、层黏连蛋白(LN)和纤连蛋白(FN)的含量。结果与空白对照组比较,CysC高表达组中CatS的表达较少,差异无统计学意义(P>0.05);随干扰时间延长,干扰组中CatS的表达明显增强(P<0.01);Ⅰ型胶原含量在各组虽有轻微增加或减少,但差异均无统计学意义(P>0.05);LN和FN含量在CysC高表达24h组和48h组均有不同程度增加,LN含量在干扰72h组明显减少,FN含量在干扰48h组和72h组明显减少(P均<0.05)。结论正常ASMC中CatS表达极少;CysC和CatS在ASMC中的表达呈负相关;CysC和CatS的平衡关系对层黏连蛋白和纤连蛋白均有不同程度的影响,但对Ⅰ型胶原的影响不明显。
Objective To study the effect of Cystatin C on Cathepsin S and extracellular matrix（ECM）in human arterial smooth muscle cells（ASMCs）.Methods ASMCs were divided into the control group（normal cells）,the Cystatin C highly expressed group,the Cystatin C silenced group and the independent interference group.Expression of Cathepsin S was detected by RT-PCR and Western blot at different times.Contents of collagenⅠ,laminin（LN）and fibronectin（FN）were detected by ELISA in ASMCs of the control group,the Cystatin C highly expressed group（24 h and 48 h）and the Cystatin C silenced group（48 h and 72 h）.Results Expression of Cathepsin S in the Cystatin C highly expressed group was lower than in the control group,and the difference between the two groups was not statistically significant（P〉0.05）.Expression of Cathepsin S in the Cystatin C silenced group was obviously strengthened with the increase of interference time（P〈0.01）.At the same time,expression of Cystatin C was weakened.The result of ELISA showed that content of collagenⅠ in each group was increased or decreased,while the differences were not statistically significant（P〉0.05）.Contents of LN and FN increased in the Cystatin C highly expressed group（24 h and 48 h）,and the differences were both statistically significant compared with the controls（P〈0.05）.Compared with the control group,content of LN in Cystatin C silenced group（48 h）was insignificantly decreased（P〉0.05）,while it was obvi-ously decreased in the Cystatin C silenced group（72 h）（P〈0.05）.Compared with the control group,contents of FN in the Cystatin C silenced group（48 h and 72 h）markedly decreased,and the differences were both statistically significant（P〈0.05）.Conclusion Expression of Cathepsin S is very low in normal ASMCs.Expressions of Cystatin C and Cathepsin S in ASMCs are inversely correlated.The balance of Cystatin C and Cathepsin S has a significant effect on LN and FN to different degrees,but no significant effec
出处
《山东大学学报（医学版）》
CAS
北大核心
2010年第12期124-128,133,共6页
Journal of Shandong University:Health Sciences
