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机构地区:[1]解放军第451医院心血管内科,陕西西安710054
出 处:《生物技术通讯》2014年第2期234-236,共3页Letters in Biotechnology
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
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