丹参素对缺氧诱导成纤维样滑膜细胞损伤的影响及作用机制研究  

Effect and Mechanism of Danshensu on Hypoxia-induced Fibroblast-like Synovial Cell Injury

作　　者：杨帆[1] 曹姗 吕章明 李云海[1] YANG Fan;CAO Shan;LYU Zhangming;LI Yunhai(Jingui Teaching and Research Section,Hubei University of Chinese medicine,Wuhan 430061,China)

机构地区：[1]湖北中医药大学金匮教研室,武汉430061

出　　处：《世界中医药》2024年第23期3625-3631,共7页World Chinese Medicine

基　　金：国家中医药管理局“田玉美全国名老中医药专家传承工作室”项目[(2012)149];湖北省教育厅科学研究计划指导性项目(B2021116)。

摘　　要：目的:探讨丹参素对缺氧诱导的成纤维样滑膜细胞损伤的作用机制。方法:将大鼠成纤维样滑膜细胞按照简单随机法分为对照组、缺氧组、低剂量组、中剂量组、高剂量组,检测丹参素对成纤维样滑膜细胞的影响。另取同代大鼠细胞按照简单随机法分为对照组、缺氧组、高剂量组、BAY11-7082组,检测核因子κB p65(NF-κB p65)抑制剂BAY11-7082对成纤维样滑膜细胞损伤的影响。以5-乙炔基-2′-脱氧尿苷(Edu)法检测各组细胞增殖,蛋白质印迹法检测细胞蛋白表达水平,实时荧光定量聚合酶链反应(RT-qPCR)检测各组细胞细胞基质金属蛋白酶(MMP)及血管内皮生长因子(VEGF)mRNA表达。结果:缺氧组细胞Edu阳性细胞率、增殖细胞核抗原(Ki67)、细胞周期素蛋白D1(CyclinD1)蛋白表达和MMP-1、MMP-9、MMP-3、VEGF mRNA及磷酸化(p)-NF-κB p65、低氧诱导因子1α(HIF-1α)、白细胞介素-6(IL-6)、IL-8蛋白表达高于对照组;与缺氧组比较,低、中、高剂量组细胞Edu阳性细胞率及Ki67、PCNA、CyclinD1蛋白表达及MMP-1、MMP-9、MMP-3、VEGF mRNA、p-NF-κB p65、HIF-1α、IL-6、IL-8蛋白表达低(P<0.05);与高剂量组比较,BAY11-7082组细胞p-NF-κB p65、HIF-1α、IL-6、IL-8蛋白表达、Edu阳性细胞率及MMP、VEGF mRNA低(P<0.05)。结论:丹参素可能通过抑制NF-κB/HIF-1α信号通路改善缺氧诱导的大鼠成纤维样滑膜细胞损伤。Objective:To explore the mechanism by which Danshensu protects against hypoxia-induced injury in fibroblast-like synovial cells.Methods:Rat fibroblast-like synovial cells were randomly divided into control,hypoxia,low-dose,medium-dose,and high-dose groups.The effect of Danshensu on fibroblast-like synovial cells were observed.Additionally,cells from the same generation of rats were divided into control,hypoxia,high-dose,and BAY11-7082 groups to assess the effect of the NF-κB p65 inhibitor BAY11-7082 on fibroblast-like synovial cell injury.Cell proliferation was detected by 5-ethynyl-2′-deoxyuridine(Edu)assay,protein expression levels were analyzed using Western blot,and mRNA expression of matrix metalloproteinases(MMPs)and vascular endothelial growth factor(VEGF)was measured by RT-qPCR.Results:In the hypoxia group,Edu-positive cell rate,Ki67,CyclinD1 protein expression,MMP-1,MMP-9,MMP-3,VEGF mRNA,and phosphorylated(p)-NF-κB p65,hypoxia-inducible factor 1α(HIF-1α),interleukin-6(IL-6),and IL-8 protein levels were higher than the control group.Compared with the hypoxia group,low,medium,and high-dose groups showed significantly reduced Edu-positive cell rates,Ki67,PCNA,CyclinD1 protein expression,MMP-1,MMP-9,MMP-3,VEGF mRNA,p-NF-κB p65,HIF-1α,IL-6,and IL-8 protein expression(P<0.05).Compared to the high-dose group,BAY11-7082 treatment further decreased the expression of p-NF-κB p65,HIF-1α,IL-6,IL-8,Edu-positive cell rate,and MMP and VEGF mRNA levels(P<0.05).Conclusion:Danshensu may alleviate hypoxia-induced fibroblast-like synovial cell injury in rats by inhibiting the NF-κB/HIF-1αsignaling pathway.

关 键 词：类风湿关节炎 丹参素 缺氧 成纤维样滑膜细胞 核因子κB/低氧诱导因子1α信号通路 细胞增殖 细胞损伤 大鼠 

分 类 号：R285[医药卫生—中药学]