红景天苷通过激活PI3K/Akt和抑制NF⁃κB信号通路改善川崎病冠状动脉损伤  

Salidroside improves coronary artery injury in Kawasaki disease by activating PI3K/Akt and inhibiting NF⁃κB signaling pathway

作　　者：胡琳 杨艳娟 李洁滢 周忠 田正 焦蓉 HU Lin;YANG Yanjuan;LI Jieying;ZHOU Zhong;TIAN Zheng;JIAO Rong(Xiangyang No.1 People′s Hospital,Hubei University of Medicine,Xiangyang 441000,China)

机构地区：[1]湖北医药学院附属襄阳市第一人民医院儿科,湖北襄阳441000

出　　处：《实用医学杂志》2022年第21期2657-2662,2669,共7页The Journal of Practical Medicine

基　　金：湖北省儿科联盟医学科研项目(编号:HPAMRP202108);襄阳市科学技术局项目(编号:2020ZD34)。

摘　　要：目的观察红景天苷(Salidroside,Sal)对川崎病冠状动脉炎症反应和氧化应激的影响并探讨其作用机制。方法将细胞分为正常对照组、TNF⁃α组、50μmol/L Sal组和100μmol/L Sal组。用CCK⁃8法检测不同浓度Sal和肿瘤坏死因子⁃α(TNF⁃α)对HCAEC增殖活性的影响;通过Western blot法检测各组细胞中磷酸化磷脂酰肌醇3激酶(p⁃PI3K)、磷酸化蛋白激酶B(p⁃Akt)、磷酸化核转录因子⁃κBp65(p⁃P65)、白细胞介素6(IL⁃6)、白细胞介素⁃1β(IL⁃1β)、紧密连接蛋白(ZO⁃1)及血管内皮钙黏蛋白(VE⁃cadherin)的蛋白表达水平;通过RT⁃qPCR法检测各组炎症因子和黏附分子mRNA的表达水平;利用DCFH⁃DA荧光探针观察细胞内活性氧(ROS)的生成情况;分光光度仪检测细胞内超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH⁃Px)的含量。结果与对照组相比,TNF⁃α组p⁃PI3K、p⁃Akt、ZO⁃1、VE⁃cadherin的蛋白表达水平降低(P<0.05),p⁃P65、IL⁃6和IL⁃1β蛋白表达水平升高(P<0.05),IL⁃6、IL⁃8、IL⁃1β、ICAM⁃1、VCAM⁃1的mRNA表达水平显著升高(P<0.05),ROS的生成增加,SOD和GSH⁃Px的活性显著下降,而Sal可显著逆转上述作用(P<0.05)。结论Sal能减轻TNF⁃α诱导的HCAEC细胞炎症反应和氧化应激水平,其机制可能与激活PI3K/Akt和抑制NF⁃κB信号通路有关。Objective To study the effect of salidroside(Sal)on the inflammatory response and oxidative stress of coronary arteries in Kawasaki disease and its mechanism.Methods The cells were divided into normal control group,TNF⁃αgroup,50μmol/L and 100μmol/L Sal group.The effect of Sal and TNF⁃αat different concentrations on HCAEC survival was detected by CCK⁃8.The relative protein expression levels of phosphorylated phosphatidylinositol 3 kinase(p⁃PI3K),phosphorylated protein kinase B(p⁃Akt),phosphorylated NF⁃κBp65(p⁃P65),interleukin 6(IL⁃6),IL⁃1β,tight junction protein(ZO⁃1)and vascular endothelial calherin(VE⁃cad⁃herin)were detected by Western blot.The expression levels of inflammatory factors and adhesion molecule mRNAs were detected by RT⁃qPCR.Intracellular reactive oxygen species(ROS)production was observed by DCFH⁃DA fluorescent probes.Intracellular superoxide dismutase(SOD)and glutathione peroxidase(GSH⁃Px)were analyzed with spectrophotometers.Results Compared with the control group,the protein expression levels of p⁃PI3K,p⁃Akt,ZO⁃1 and VE⁃cadherin were significantly reduced(P<0.05),the protein expression levels of p⁃P65,IL⁃6 and IL⁃1βwere significantly increased(P<0.05),and the mRNA expression levels of IL⁃6,IL⁃8,IL⁃1β,ICAM⁃1,and VCAM⁃1 were significantly increased(P<0.05).The generation of ROS was significantly increased and the viability of SOD and GSH⁃Px were significantly decreased in the TNF⁃αgroup(P<0.05),while Sal could significantly reverse these effects.Conclusion Salidroside can attenuate TNF⁃α⁃induced inflammatory response and oxidative stress levels in HCAEC cells probably by way of activating PI3K/Akt and inhibiting NF⁃κB signal⁃ing pathway.

关 键 词：川崎病 红景天苷 炎症反应 氧化应激 肿瘤坏死因子⁃α 人冠状动脉内皮细胞 

分 类 号：R725.4[医药卫生—儿科] R363.2[医药卫生—临床医学]