【摘要】 [目的]探讨瞬时受体电位香草酸亚型1（TRPV1）对Ca²⁺依赖性转录调节因子表达的影响以及大鼠血管平滑肌细胞（RVSMC）增殖的作用。[方法]体外培养RVSMC,利用TRPV1特异性激动剂辣椒素（CAP）或抑制剂辣椒卓平（CPZ）促进或抑制TRPV1的表达。免疫细胞化学染色技术检测TRPV1的表达；噻唑蓝法检测细胞增殖活性；Western blot检测增殖细胞核抗原（PCNA）蛋白表达；Western blot和实时荧光定量PCR检测Ca²⁺依赖性转录调节因子活化T细胞核因子c1（NFATc1）、钙衰蛋白（CSEN）和肌细胞增强因子2C（MEF2C）的蛋白表达和mRNA水平。[结果]免疫细胞化学染色显示TRPV1主要表达在RVSMC的胞膜和胞质，CAP或CPZ能激活或抑制TRPV1的表达。与对照组相比，CAP可明显刺激RVSMC增殖活性和上调PCNA蛋白的表达（P<0.01）,同时，NFATc1、MEF2C的蛋白和mRNA水平显著上调（P<0.01）;CSEN蛋白表达显著增高（P<0.01）,CSEN mRNA水平无明显变化（P>0.05）。CPZ作用后，与对照组比较，RVSMC增殖活性、PCNA蛋白表达降低（P<0.01）,NFATc1、MEF2C的蛋白和mRNA水平显著下调（P<0.01）;CSEN蛋白表达显著降低（P<0.01）,CSEN mRNA水平无明显变化（P>0.05）。[结论] TRPV1可能通过上调Ca²⁺依赖性转录调节因子的表达参与促RVSMC增殖。更多还原

【Abstract】 Aim To investigate the effect of transient receptor potential vanilloid subtype 1（TRPV1） on the expression of Ca²⁺-dependent transcriptional regulator and the proliferation of rat vascular smooth muscle cell（RVSMC）.Methods RVSMCs were cultured in vitro,and TRPV1-specific agonist capsaicin（ CAP） or inhibitor capsazepine（ CPZ）was used to promote or inhibit the expression of TRPV1. TRPV1 expression was detected by immunocytochemical staining technique. Cell proliferation activity was detected by methyl thiazolyl tetrazolium method. Proliferating cell nuclear antigen（ PCNA） protein expression was detected by Western blot. Western blot and quantitative real-time PCR were used to detect the protein expression and m RNA level of Ca²⁺-dependent transcriptional regulator such as nuclear factor of activated T cell c1（ NFATc1）,calsenilin（ CSEN） and myocyte enhancer factor 2C（ MEF2C）. Results Immunocytochemical staining showed that TRPV1 was mainly expressed in the membrane and cytoplasm of RVSMC,and CAP or CPZ could activate or inhibit the expression of TRPV1. Compared with the control group,CAP could significantly stimulate the proliferation activity of RVSMCs and up-regulate the expression of PCNA protein（ P<0. 01）. Meanwhile,the protein and m RNA levels of NFATc1 and MEF2C were significantly up-regulated（ P<0. 01）; The protein expression of CSEN was significantly increased（ P<0. 01）,and there was no significant change in CSEN m RNA level（ P>0. 05）. After CPZ treatment,compared with the control group,the proliferation activity of RVSMC and the expression of PCNA protein were decreased（ P<0. 01）,the protein and m RNA levels of NFATc1 and MEF2C were significantly down-regulated（ P< 0. 01）; The protein expression of CSEN was significantly decreased（ P<0. 01）,and there was no significant change in CSEN m RNA level（ P>0. 05）. Conclusion TRPV1 may be involved in promoting RVSMC proliferation by up-regulating the expression of Ca²⁺-dependent transcriptional regulator.更多还原