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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 investigate the effects of cystatin C (CysC) on cathepsin S (CatS) and extracellular matrix in human arterial smooth muscle cells (ASMCs). Methods CysC (interference group) 0, 12, and 24 were respectively transfected with high expression of CysC plasmid (high CysC expression group) and small interfering RNA (siRNA) by RT-PCR and Western- The expression of CatS was detected at 48 and 72h, the expression of collagen Ⅰ, laminin (LN) and fibronectin (FN) in 48h and 72h group were significantly higher than those in control group content. Results Compared with the blank control group, the expression of CatS in the CysC high expression group was less (P> 0.05), and the expression of CatS in the interference group was significantly increased with the prolongation of the interference time (P <0.01) Collagen content increased slightly or decreased in each group, but the differences were not statistically significant (P> 0.05). The contents of LN and FN in CysC high expression 24h group and 48h group were increased in varying degrees, LN content in the intervention 72h group Significantly decreased, FN content in the disturbance 48h group and 72h group was significantly reduced (P all <0.05). Conclusions The expression of CatS in normal ASMC is extremely low. The expression of CysC and CatS in ASMC is negatively correlated. The equilibrium relationship between CysC and CatS has different effects on laminin and fibronectin, but the effect on type I collagen Not obvious.