红景天苷通过抑制TLR 4/NF-κB通路抑制高磷诱导的血管平滑肌细胞钙化  

Salidroside Attenuates High Phosphate-induced Calcification of Vascular Smooth Muscle Cells Via Inhibiting TLR 4/NF-κB Pathway

作　　者：刘芳[1] 李明熹 颜建云 LIU Fang;LI Mingxi;YAN Jianyun(Department of Cardiology,Laboratory of Heart Center,Zhujiang Hospital,Southern Medical University,Guangzhou 510280,China;Department of Pathophysiology,Zhongshan School of Medicine,Sun Yat-Sen University,Guangzhou 510080,China)

机构地区：[1]南方医科大学珠江医院心血管内科,珠江医院心脏中心实验室,广东广州510280 [2]中山大学中山医学院病理生理学教研室,广东广州510080

出　　处：《中山大学学报（医学科学版）》2024年第6期883-890,共8页Journal of Sun Yat-Sen University:Medical Sciences

基　　金：国家自然科学基金(82070246)。

摘　　要：【目的】本文旨在探讨红景天苷对血管平滑肌细胞钙化的作用及机制。【方法】本研究采用人血管平滑肌细胞钙化模型,用不同浓度的红景天苷处理细胞,茜素红染色及钙离子浓度定量检测细胞钙化,Western blot检测红景天苷对成骨样分化蛋白Runt相关转录因子2(RUNX2)和骨形态发生蛋白2(BMP2)的影响;检测红景天苷对Toll样受体4(TLR4)、核因子κB p65(NF-κB p65)和磷酸化的核因子κB p65(p-NF-κB p65)的影响;检测红景天苷对白细胞介素6(IL-6)、白细胞介素1β(IL-1β)的影响。采用TLR4小干扰RNA(si-TLR4)处理血管平滑肌细胞,研究TLR4介导红景天苷对平滑肌细胞钙化的抑制作用。【结果】茜素红染色及钙离子浓度定量结果提示不同浓度的红景天苷改善高磷诱导的人血管平滑肌细胞的钙化(P<0.05)。与高磷模型组相比,红景天苷(250μmol/L、500μmol/L)下调成骨样分化蛋白RUNX2(P<0.05)和BMP2(P<0.05);250μmol/L、500μmol/L浓度的红景天苷还下调高磷诱导的TLR4(P<0.05)和p-NF-κB p65(P<0.05)的表达,而对NF-κB p65影响无显著差异(P>0.05);红景天苷(250μmol/L、500μmol/L)抑制炎症因子IL-1β(P<0.05)、IL-6(P<0.05)的蛋白水平。此外,茜素红染色及钙定量提示,红景天苷可增强敲低TLR4对高磷诱导的血管平滑肌细胞钙化的抑制作用(P<0.05)。【结论】红景天苷可通过抑制TLR4/NF-κB通路抑制高磷诱导的血管平滑肌细胞钙化。【Objective】To investigate the effect of salidroside on calcification of vascular smooth muscle cells(VSMCs)and explore its underlying mechanism.【Methods】VSMCs were cultured in high-phosphate media to induce vascular calcification and treated with different concentrations of salidroside.Cell calcification was tested by alizarin red staining and calcium content assay.qRT-PCR and western blotting were used to determine the expression levels of osteogenic proteins including runt-related transcription factor 2(RUNX2)and bone morphogenetic protein 2(BMP2),toll-like receptor 4(TLR 4),nuclear factor kappa-B p 65(NF-κB p 65),phosphorylated nuclear factor kappa-B p 65(p NFκB p 65),inflammatory factors including Interleukin-1 beta(IL-1β)and Interleukin-6(IL-6).We transfected VSMCs with TLR 4 small interfering RNA(si-TLR 4)and examined whether TLR 4 mediated the inhibitory effect of salidroside on calcification of VSMCs.【Results】Alizarin red staining and calcium content assay revealed that different concentrations of salidroside ameliorated high phosphate-induced calcification of human VSMCs(P<0.05).Salidroside at concentrations of 250 and 500μmol/L decreased the expression levels of RUNX 2,BMP 2,TLR 4,p-NF-κB p 65,IL-1βand IL-6(all P<0.05),but did not affect NF-κB p 65 expression(P>0.05).The examination of VSMCs transfected with si-TLR 4 showed that salidroside enhanced the inhibitory effect of TLR 4 knockdown on calcification of VSMCs(P<0.05).【Conclusions】Salidroside attenuates high phosphate-induced calcification of VSMCs via inhibiting TLR 4/NF-κB pathway.

关 键 词：红景天苷 血管平滑肌细胞 血管钙化 炎症 NF-ΚB 

分 类 号：R543[医药卫生—心血管疾病]