葫芦茶苷调控NF-κB/NLRP3/Caspase-1信号通路抑制肝星状细胞活化的作用机制  

Study on the mechanism of tadehaginoside inhibiting the activation of hepatic stellate cells based on the NF-κB/NLRP3/Caspase-1 signal pathway

作　　者：卢秋玉[1] 韦璐莹 李鑫[3] 夏冰雨 唐爱存[2] Lu Qiu-yu;Wei Lu-ying;Li Xin;Xia Bing-yu;Tang Ai-cun(Department of Pharmacy,People's Hospital of Guangxi Zhuang Autonomous Region,Nanning,Guangxi 530021,China;The First Affiliated Hospital of Guangxi Traditional Chinese Medical University,Nanning,Guangxi 530023,China;Guangxi University of Chinese Medicine,Nanning,Guangxi 530200,China)

机构地区：[1]广西壮族自治区人民医院药学部,广西南宁530021 [2]广西中医药大学第一附属医院,广西南宁530023 [3]广西中医药大学,广西南宁530200

出　　处：《中国现代医学杂志》2025年第7期48-53,共6页China Journal of Modern Medicine

基　　金：广西自然科学基金(No:2023GXNSFAA026199);广西中医药管理局科研课题(No:GXZYA20220180,GXZYZ20210209);广西中医药大学校级科研项目(No:2021MS015)。

摘　　要：目的探讨葫芦茶苷调控NF-κB/NLRP3/Caspase-1信号通路抑制肝星状细胞(HSCs)活化的作用机制。方法制备人HSCs的体外细胞(LX-2细胞)模型,采用CCK-8法检测葫芦茶苷对LX-2增殖的影响。将对数生长期的LX-2细胞随机分成5组:空白组、TGF-β_(1)活化组、TGF-β_(1)+葫芦茶苷低剂量组、TGF-β_(1)+葫芦茶苷中剂量组、TGF-β_(1)+葫芦茶苷高剂量组,除空白组外,其余各组分别加入含终浓度为5μg/L的TGF-β_(1)的培养液,使其增殖活化24 h,再对葫芦茶苷低、中、高剂量组加入终浓度分别为20、40和80μg/mL的葫芦茶苷进行干预。CCK-8法检测葫芦茶苷对LX-2细胞增殖的影响。酶联免疫吸附试验检测细胞上清液白细胞介素-1β(IL-1β)、白细胞介素-18(IL-18)、白细胞介素-6(IL-6)、干扰素α(INF-α)表达水平。实时荧光定量聚合酶链反应和Western blotting实验检测细胞中NF-κB、NLRP3、Caspase-1基因和蛋白相对表达量。结果葫芦茶苷能显著抑制LX-2细胞的增殖。与空白组比较,TGF-β_(1)活化组细胞上清液IL-1β、IL-18、IL-6、INF-α表达水平,以及细胞NF-κB、NLRP3、Caspase-1基因和蛋白相对表达量明显上升(P<0.05)。葫芦茶苷干预后,与TGF-β_(1)活化组比较,葫芦茶苷不同剂量组降低细胞上清液IL-1β、IL-18、IL-6、INF-α表达水平,下调细胞NF-κB、NLRP3、Caspase-1基因和蛋白表达(P<0.05)。结论葫芦茶苷能显著抑制HSCs增殖活化,其机制可能与调控NF-κB/NLRP3/Caspase-1信号通路有关。Objective To explore the mechanism of Tadehaginoside in regulating the NF-κB/NLRP3/Caspase-1 signaling pathway and inhibiting hepatic stellate cell activation.Methods An in vitro cell model of human hepatic stellate cell LX-2 was developed,and using CCK-8 method to detect the effect of Tadehaginoside on LX-2 proliferation.Randomly divide LX-2 cells in logarithmic growth phase into 5 groups:a blank control group,a TGF-β_(1)activation group,a TGF-β_(1)+Tadehaginoside low-dose group,a TGF-β_(1)+Tadehaginoside medium-dose group,and a TGF-β_(1)+Tadehaginoside high-dose group.All other groups except for the blank group were added to a culture medium containing TGF-β_(1)(final concentration of 5μg/L)to promote proliferation and activation for 24 hours.Then,low,medium,and high-dose Tadehaginoside groups(final concentrations of 20μg/mL,40μg/mL,and 80μg/mL)were added for intervention.The CCK-8 method was used to detect the effect of Tadehaginoside on LX-2 proliferation.ELISA method detected the expression levels of IL-1β,IL-18,IL-6,and INF-αin the cell supernatant.qRT-PCR and Western blot were used to detect the mRNA and protein expression levels of NF-κB,NLRP3,and Caspase-1 in cells.Results Tadehaginoside can significantly inhibit the proliferation of LX-2 cells.Compared with the blank group,the TGF-β_(1)activation group significantly increased the expression levels of IL-1β,IL-18,IL-6,and INF-αin the cell supernatant,as well as the mRNA and protein expression levels of NF-κB,NLRP3,and Caspase-1 in the cells,with statistical differences(P<0.05).After intervention with Tadehaginoside,compared with the TGF-β_(1)activation group,various doses of Tadehaginoside could reduce the expression levels of IL-1β,IL-18,IL-6,and INF-αin cell supernatant and downregulate the mRNA and protein expression of NF-κB,NLRP3,and Caspase-1 in cells,with statistical differences(P<0.05).Conclusions Tadehaginoside can significantly inhibit the proliferation and activation of hepatic stellate cells,and its mechanism may be relat

关 键 词：肝纤维化 葫芦茶苷 肝星状细胞 NF-κB/NLRP3/Caspase-1通路 

分 类 号：R575.2[医药卫生—消化系统]