黄芩苷调节let-7i-3p/PI3K/Akt/NF-κB信号轴减轻类风湿关节炎成纤维样滑膜细胞NLRP3炎性小体活化  

Baicalin modulates let-7i-3p/PI3K/Akt/NF-κB signaling axis to reduce NLRP3 inflammasome activation in fibroblast like synoviocytes of rheumatoid arthritis

作　　者：张炜 王莉[2] 杨雨欣 马锐 王丽 黄菱 万巧凤 ZHANG Wei;WANG Li;YANG Yu-xin;MA Rui;WANG Li;HUANG Ling;WAN Qiao-feng(Dept of Pathogenic Biology and Immunology,School of Basic Medicine,Ningxia Medical University,Yinchuan 750004,China;Dept of Rheumatology,Ningxia Medical University General Hospital,Yinchuan 750003,China;School of Clinical Medicine,Ningxia Medical University,Yinchuan 750004,China)

机构地区：[1]宁夏医科大学基础医学院病原生物学与医学免疫学系,宁夏银川750004 [2]宁夏医科大学总医院风湿科,宁夏银川750003 [3]宁夏医科大学临床医学院,宁夏银川750004

出　　处：《中国药理学通报》2023年第12期2313-2319,共7页Chinese Pharmacological Bulletin

基　　金：宁夏自然科学基金资助项目(No 2022AAC03152)。

摘　　要：目的研究黄芩苷减轻类风湿关节炎成纤维样滑膜细胞(human fibroblast like synoviocytes of rheumatoid arthritis,HFLS-RA)NLRP3炎性小体活化的作用及机制。方法为证实黄芩苷减轻HFLS-RA中NLRP3炎性小体活化,采用免疫荧光观察黄芩苷处理前后NLRP3的表达;Western blot检测黄芩苷处理48 h后,p-PI3K、p-Akt、NF-κB p65、NLRP3、ASC及caspase-1蛋白的表达;ELISA检测细胞上清中IL-1及IL-18的含量。为探究黄芩苷减轻NLRP3炎性小体活化的机制,采用双荧光素酶及Western blot验证let-7i-3p与PIK3CA的对应关系;RT-qPCR检测黄芩苷干预前后let-7i-3p及PI3K的表达;采用let-7i-3p干扰,验证黄芩苷是否减轻了增强的NLRP3炎性小体的活化。结果50、100 mg·L^(-1)黄芩苷明显减轻了NLRP3炎性小体的活化,抑制p-PI3K、p-Akt、NF-κB p65、NLRP3、ASC、caspase-1蛋白的表达,抑制IL-1、IL-18的分泌。let-7i-3p与PIK3CA存在靶向对应关系,黄芩苷上调了let-7i-3p的表达、下调了PIK3CA的表达;黄芩苷减轻了因let-7i-3p干扰而增强的NLRP3炎性小体的活化。结论黄芩苷经上调let-7i-3p表达、抑制PI3K/Akt/NF-κB信号转导,从而减轻NLRP3炎性小体的活化。Aim To study the effect of baicalin on the activation of NLRP3 inflammasomes in human fibroblast like synoviocytes of rheumatoid arthritis(HFLS-RA)and its mechanism.Methods To confirm that baicalin alleviated the activation of NLRP3 inflammasome in HFLS-RA,immunofluorescence was used to observe the expression of NLRP3 before and after baicalin treatment.Western blot was used to detect the protein expression of p-PI3K,p-Akt,NF-κB p65,NLRP3,ASC and caspase-1 after baicalin treatment for 48 h,and ELISA was employed to detect the contents of IL-1 and IL-18 in the supernatents.In order to explore the mechanism of baicalin alleviating the activation of NLRP3 inflammasome,double luciferin and Westen blot analysis were applied to verify the corresponding relationship between let-7i-3p and PIK3CA.RT-qPCR was utilized to determine the expression of let-7i-3p and PI3K before and after baicalin intervention.let-7i-3p interference was used to verify whether baicalin mitigated the activation of enhanced NLRP3 inflammasomes.Results Baicalin(50,100 mg·L^(-1))significantly reduced the activation of NLRP3 inflammasomes,inhibited the protein expressions of p-PI3K,p-Akt,NF-κB p65,NLRP3,ASC and caspase-1,and the secretion of IL-1 and IL-18.let-7i-3p and PIK3CA had a targeted correspondence,and baicalin up-regulated the expression of let-7i-3p and down-regulated the expression of PIK3CA.Baicalin attenuated the activation of NLRP3 inflammasomes enhanced by let-7i-3p interference.Conclusions Baicalin can up-regulate let-7i-3p expression,inhibit PI3K/Akt/NF-κB signal transduction,and thus reduce the activation of NLRP3 inflammasomes in HFLS-RA.

关 键 词：黄芩苷 类风湿关节炎 类风湿关节炎成纤维样滑膜细胞 NLRP3炎性小体 miRNA 双荧光素酶 

分 类 号：R284.1[医药卫生—中药学] R342.2[医药卫生—中医学] R364.5R593.22R977.3