摘要
目的:研究Caspase-1抑制剂zVAD对大鼠血管平滑肌细胞钙化的影响。方法:体外分离、培养大鼠血管平滑肌细胞,加入β-磷酸甘油酯(β-GP)诱导细胞发生钙化;在细胞中加入Caspase-1抑制剂zVAD或DMSO对照,按照不同时间点收取细胞进行半定量PCR,检测骨保护素(OPG)/核因子κB受体活化因子配体(RANKL)的表达变化;通过茜素红钙化染色,直观观察zVAD对血管平滑肌细胞钙化的影响。结果:β-GP加入细胞后,可见细胞内OPG/RANKL的mRNA表达水平逐步增加,加入zVAD后增加趋势减缓;茜素红钙化染色显示,zVAD可抑制血管平滑肌细胞钙化。结论:分离、体外培养了大鼠的血管平滑肌细胞,并且发现在钙化过程中OPG/RANKL表达增加,而Caspase-1抑制剂zVAD可有效抑制OPG/RANKL的表达,提示炎症小体可能通过OPG/RANKL诱导动脉钙化的产生。
Objective： To explore the effect of caspase-1 inhibitor zVAD on rat vascular smooth muscle cells （VSMC） in the process of calcification. Methods： To build calcification model in vitro, we added β-glycerol phos phate（β-GP） to VSMC isloted from SD rat. Semi-quantative PCR was used to detect the expression levels of os teoprotegerin（OPG） and receptor activator of nuclear factor KB ligand（RANKL）. Alizarin Red S stain assay was used to evaluate the level of arterial calcification. Results： mRNA level of OPG and RANKL was increased dur ing VSMC calcification, however this increasing was downregulated by caspase-1 inhibitor zVAD. In the meantime, Alizarin Red S stain showed that calcification was also improved with additon of zVAD. Conclusion： These re suits suggest that inflammasome activation might affect VSMC calcification through OPG/RANKL modulation
出处
《生物技术通讯》
CAS
2014年第2期234-236,共3页
Letters in Biotechnology
