白藜芦醇调节TRPC1/STIM1介导的SOCC抗动脉粥样硬化机制

Effect of Resveratrol in Regulating TRPC1/STIM1-mediated SOCC to Inhibit Atherosclerosis

目的：研究白藜芦醇通过调节瞬时受体电位通道蛋白1（transient receptor potential channel protein 1,TRPC1）及基质交互分子1（stromal interaction molecule 1,STIM1）介导的钙库操纵性钙通道（store-operated calcium channel,SOCC）发挥抗动脉粥样硬化（atherosclerosis,AS）的具体机制。方法：雌性C57BL/6J小鼠卵巢去势后高脂饮食建立AS模型,分为空白组,模型组,白藜芦醇低、中、高剂量组（50,100,150 mg·kg^-1）,雌激素组（0.3 mg·kg^-1）。高脂喂养16周后,收集血清检测各组小鼠血清中血脂4项及Ca^2＋水平;油红O染色从形态学观察动脉粥样硬化斑块形成;蛋白免疫印迹法（Western blot）检测小鼠胸主动脉TRPC1,STIM1及内皮型一氧化氮合成酶（endothelial nitric oxide synthase,eNOS）蛋白表达水平;实时荧光定量PCR法（Realtime PCR）检测小鼠胸主动脉血管中TRPC1与STIM1 mRNA信号表达。结果：与模型组相比,白藜芦醇高剂量组及雌激素组小鼠血清总胆固醇（TC）,甘油三酯（TG）,低密度脂蛋白（LDL）均降低（P〈0.05）,但高密度脂蛋白表达（HDL）升高,血清Ca^2＋浓度下降;油红O染色提示,与空白组相比,模型组AS病理改变明显,内膜增厚,不完整,连续性被破坏,有较明显的脂质斑块;补充白藜芦醇或雌激素后,动脉管壁AS病理改变明显减轻,脂滴明显减少。Western blot结果显示,补充白藜芦醇中、高剂量及雌激素后,与模型组相比,TRPC1与STIM1蛋白表达明显减少,而eNOS蛋白表达增加（P〈0.05）;通过Real-time PCR检测发现,与模型组相比,白藜芦醇中、高剂量组及雌激素组TRPC1与STIM1 mRNA基因表达的水平明显降低（P〈0.05）。结论：白藜芦醇可通过下调TRPC1及STIM1基因表达及蛋白水平,抑制SOCC介导的Ca^2＋内流,增加eNOS蛋白表达及活性,发挥抗动脉粥样硬化的作用。

Objective： To study the anti-atherosclerosis（AS） mechanism of resveratrol（RES） by regulating store-operated calcium channel（SOCC） induced by transient receptor potential channel protein 1（TRPC1） and stromal interaction molecule 1（STIM1）.Method： Atherosclerosis model was established with female C57 BL/6 J mice that were fed with high-fat diet after ovariectomy,and then the mice were divided into control group,model group,low-dose resveratrol group,medium-dose resveratrol group,high-dose resveratrol group（50,100,150 mg·kg^-1） and estradiol（E2） group（0.3 mg·kg^-1）.After 16 weeks,serum was collected,and the lipid level and Ca^2＋concentration were detected.The morphological changes were measured by oil red O staining.The protein expressions of TRPC1,STIM1 and endothelial nitric oxide synthase（eNOS） were detected by Western blot and Real-time PCR.The activity of eNOS was measured by enzyme-linked immunosorbent assay（ELISA）.Result： Compared with model group,total cholesterol（TC）,triglyceride（TG）,low density lipoprotein（LDL） were decreased,while the high density lipoprotein（HDL） was increased in supplementary high-dose resveratrol or estrogen groups（P〈0.05）.The pathological changes in the model group were obvious compared with control group,after supplementation with RES or estrogen,the pathological changes were significantly reduced in thoracic aorta.Western blot and Real-time PCR results showed that in model group,the protein expressions of TRPC1 and STIM1 were increased,while the protein expression of eNOS was decreased（P〈0.05）; in supplementary high-dose resveratrol or estrogen group,these changes were reversed（P〈0.05）.Conclusion： RES could down-regulate TRPC1 and STIM1 gene and protein expressions,inhibit SOCC-mediated Ca^2＋ influx,increase eNOS expression and resist atherosclerosis.