雷公藤红素通过调节血管平滑肌细胞表型转化改善血管重塑的作用机制研究  

Mechanism of celastrol on alleviating the vascular remodeling by modulating phenotypic transformation of vascular smooth muscle cells

作　　者：谷佳 贺卫和[1,2] 张银羽 石雅宁 邱韵 龚勇珍 覃丽[1,2,4] GU Jia;HE Wei-he;ZHANG Yin-yu;SHI Ya-ning;QIU Yun;GONG Yong-zhen;QIN Li(Laboratory of Stem Cell Regulation with Chinese Medicine and its Application,Hunan University of Chinese Medicine,Changsha 410208,Hunan Province,China;Department of Pharmacology,School of Pharmacy,Hunan University of Chinese Medicine,Changsha 410208,Hunan Province,China;Science and Technology Innovation Center,Hunan University of Chinese Medicine,Changsha 410208,Hunan Province,China;Hunan Provincial Key Laboratory of Vascular Biology and Translational Medicine,Changsha 410208,Hunan Province,China)

机构地区：[1]湖南中医药大学,干细胞中药调控与应用实验室,湖南长沙410208 [2]湖南中医药大学药学院药理学教研室,湖南长沙410208 [3]湖南中医药大学科技创新中心,湖南长沙410208 [4]血管生物学与转化医学湖南省高校重点实验室,湖南长沙410208

出　　处：《中国临床药理学杂志》2023年第14期2033-2038,共6页The Chinese Journal of Clinical Pharmacology

基　　金：国家自然科学基金项目面上基金资助项目(82274159,81973668,81774130);湖南省自然科学基金科药联合基金资助项目(2809);湖南省教育厅重点基金资助项目(20A375,22B0355);湖南省卫生健康委员会重点指导课题基金资助项目(202213055529);湖南省中医药管理局科研课题基金资助项目(B2023051);长沙市科技局科研基金资助项目(kq2004060);中药粉体与创新药物省部共建国家重点实验室培育基地开放基金资助项目(21PTKF1004);湖南中医药大学研究生创新课题立项基金资助项目(2022CX76)。

摘　　要：目的 研究雷公藤红素(CeT)通过调节血管平滑肌细胞(VSMCs)表型转化对血管重塑的作用及机制。方法 将A7r5细胞随机分为对照组(正常培养)、模型组(30 ng·mL^(-1)PDGF-BB)、CeT低剂量组(30 ng·mL^(-1)PDGF-BB+1.5μmol·L^(-1))、CeT中剂量组(30 ng·mL^(-1)PDGF-BB+2.0μmol·L^(-1))和CeT高剂量组(30 ng·mL^(-1)PDGF-BB+2.5μmol·L^(-1))。用划痕实验检测A7r5细胞迁移情况;用蛋白质印迹(Western blot)法检测细胞表型转化相关蛋白及细胞中信号转导及转录激活蛋白3(STAT3)和磷酸化STAT3(p-STAT3)蛋白表达情况。通过手术剥夺动脉方式建立小鼠股动脉导丝拉伤模型;将50只C57BL/6小鼠随机分为5组,每组10只,分别为假手术组、手术组、溶媒组(DMSO组)、低剂量CeT组(1 mg·kg^(-1))、高剂量CeT组(2 mg·kg^(-1))。以苏木精-伊红(HE)染色检测CeT对小鼠股动脉血管重塑的影响;以免疫组化染色检测CeT对VSMCs表型转化相关蛋白的影响。结果 与对照组比较,模型组24 h存活率和迁移率增加;与模型组相比,CeT中、高剂量实验组24、48 h存活率和迁移率均明显降低(均P<0.05)。与对照组相比,模型组的平滑肌肌球蛋白重链(SM-MHC)、α平滑肌肌动蛋白(α-SMA)蛋白表达水平均明显降低,骨桥蛋白(OPN)蛋白水平明显升高(均P<0.05);与模型组相比,CeT低、中、高剂量实验组的SM-MHC、α-SMA、平滑肌22α蛋白(SM22α)和p-STAT3/STAT3蛋白表达水平均明显升高,OPN蛋白水平明显降低(均P<0.05)。假手术组、手术组、DMSO组和低、高剂量CeT组股动脉SM-MHC水平分别为(41.55±2.21)%、(16.46±1.74)%、(16.15±1.96)%、(39.33±1.98)%、(38.92±5.97)%;α-SMA水平分别为(40.35±1.24)%、(10.60±5.50)%、(16.55±1.45)%、(41.47±4.05)%、(40.65±3.14)%;SM22α水平分别为(39.04±1.08)%、(7.19±0.81)%、(7.81±4.02)%、(31.20±4.86)%、(42.28±2.31)%;钙调蛋白(Calponin)蛋白水平分别为(37.21±2.15)%、(9.99±4.97)%、(11.27±3.69)%、(37.04±2.25)%、(36.27±3.21)Objective To investigate the effect and mechanism of celastrol on vascular remodeling by regulating phenotypic transformation of vascular smooth muscle cells(VSMCs).Methods A7r5 cells were randomly divided into control group(normal culture),model group(30 ng·mL^(-1)PDGF-BB),Ce T low-dose group(30 ng·mL^(-1)PDGF-BB+1.5μmol·L^(-1)),Ce T medium-dose group(30 ng·mL^(-1)PDGF-BB+2.0μmol·L^(-1))and Ce T high-dose group(30 ng·mL^(-1)PDGF-BB+2.5μmol·L^(-1)).The migration of A7r5 cells was examined by scratch assay;the expression of phenotypic transformation-related proteins and signal transducer and activator of transcription 3(STAT3)and phosphorylated STAT3(p-STAT3)proteins in cells was examined by Western blot.A mouse model of intimal hyperplasia induced by femoral artery guide wire strain was established;50 C57BL/6 mice were randomly divided into 5 groups of 10 mice each,which were sham group,operated group,lysed group(DMSO group),lowdose Ce T group(1 mg·kg^(-1)),and high-dose Ce T group(2 mg·kg^(-1)).Hematoxylin-eosin(HE)staining was used to detect the effect of Ce T on vascular remodeling in mice femoral arteries;immunohistochemical staining was used to detect the effect of Ce T on phenotypic transformation-related proteins in VSMCs.Results Compared with control group,the 24 h survival rate and migration rate of the model group increased;compared with the model group,the 24h and 48 h survival rates and migration rates of Ce T medium-dose and high-dose groups were significantly lower(all P<0.05).Compared with the control group,the expression levels of smooth muscle myosin heavy chain(SM-MHC)andαsmooth muscle actin(α-SMA)protein were significantly lower and the level of bone bridge protein(OPN)protein was significantly higher in the model group(all P<0.05);compared with model group,the SM-MHC,α-SMA,smooth muscle 22αprotein(SM22α)and p-STAT3/STAT3 protein expression levels in Ce T low-dose,medium-dose,and high-dose groups were significantly higher and OPN protein levels were significantly lower(all P<0.

关 键 词：雷公藤红素 血管重塑 内膜增生 血管平滑肌细胞 表型转化 

分 类 号：R28[医药卫生—中药学]