姜黄素通过调节NF-κB/NLRP3通路减轻LPS诱导小胶质细胞神经炎症损伤  

Curcumin alleviates LPS-induced neuroinflammatory damage of microglia by regulating the NF-κB/NLRP3 pathway

作　　者：季加翠[1] 孙春斌 罗恩丽 Ji Jiacui;Sun Chunbin;Luo Enli(Shandong Mental Health Center,Jinan 250014,China;School of Chemistry and Biological Engineering,University of Science and Technology Beijing,Beijing 100083,China;Second Department of Traditional Chinese Medicine,South China Hospital,Shenzhen University,Shenzhen 518116,China)

机构地区：[1]山东省精神卫生中心,济南250014 [2]北京科技大学化学与生物工程学院,100083 [3]广东深圳大学附属华南医院中医二科,深圳518116

出　　处：《中华细胞与干细胞杂志（电子版）》2024年第4期193-203,共11页Chinese Journal of Cell and Stem Cell（Electronic Edition）

基　　金：国家自然科学基金(82374567);广东省自然基金面上项目(2023A1515012779);深圳市科技创新委员会基础研究面上项目(JCYJ20210324093213035)。

摘　　要：目的研究姜黄素对脂多糖(LPS)诱导的神经细胞炎症的抑制作用,并探讨姜黄素抑制炎症的分子机制。方法以小胶质细胞(BV2)为研究对象,对照不做任何干预,模型组给予100 ng/mL LPS干预24 h构建神经炎症细胞模型,1、5、10μmol/L姜黄素+100 ng/mL LPS干预24 h,应用细胞免疫荧光法检测BV2细胞内白细胞介素6(IL-6)、IL-10、p-NF-κB、NLRP3、Bax和Bcl-2蛋白表达水平;应用Western blot检测NF-κB/NLRP3信号通路蛋白表达水平;采用ELISA法检测各组细胞上清肿瘤坏死因子α(TNF-α)、IL-1β和iNOS含量。给予NF-κB抑制剂BAY 11-7082后,检测BV2细胞内IL-6、IL-10、Bax和Bcl-2蛋白的表达水平。实验数据采用t检验、单因素方差分析和Dunnett-t检验方法分析。结果与对照比较,模型组炎症因子TNF-α(398.80±11.39比113.00±5.93)、IL-1β(592.40±11.32比206.70±6.81)、IL-6(1.73±0.09比0.99±0.03)、iNOS(88.39±2.14比32.48±1.71)、p-NF-κB(1.92±0.02比1.02±0.05)、NLRP3蛋白表达水平(10.33±0.61比1.02±0.05)和凋亡相关蛋白Bax表达水平(2.68±0.10比1.05±0.08)升高,IL-10蛋白表达水平(0.82±0.03比1.01±0.06)和Bcl-2蛋白表达水平(0.70±0.02比1.11±0.06)降低(P均<0.05)。Western blot结果表明,与对照比较,模型组p-NF-κB(2.65±0.06比1.04±0.05)和NLRP3蛋白表达水平(4.380±0.08比1.07±0.04)升高(P<0.001)。与模型组相比,高浓度姜黄素处理后TNF-α(207.90±7.37比398.80±11.39)、IL-1β(405.40±6.56比592.40±11.32)、IL-6(1.09±0.05比1.73±0.09)、iNOS(39.52±1.32比88.39±2.14)、p-NF-κB(01.21±0.05比1.92±0.02)和NLRP3蛋白表达水平(1.51±0.11比10.33±0.61)降低,IL-10蛋白表达水平(2.61±0.03比0.82±0.03)升高;与模型组相比,高浓度姜黄素处理后Bax蛋白表达水平(0.80±0.04比2.68±0.10)降低,Bcl-2蛋白表达水平(1.93±0.02比0.70±0.02)升高(P<0.001)。与模型组相比,NF-κB抑制剂作用后,IL-6蛋白表达水平(0.88±0.06比2.70±0.12)和Bax蛋白表达水平(1.14±0.04比3.63±0.16)降Objective To investigatethe inhibitory effect and mechanisms of curcumin on lipopolysaccharide(LPS)-induced inflammation of nerve cells.Methods Microglia(BV2)was used in current study and named control group(without any treatment),model group(treated with LPS 100 ng/mL for 24 h)and treatment group(curcumin at 1,5,10μmol/L concentration plus 100 ng/ml LPS for 24 h).The expression levels of interleukin-6(IL-6),IL-10,p-NF-κB,NLRP3,Bax and Bcl-2 in BV2 cells were detected by immunofluorescence assay.Western blot was used to detect the protein expression of NF-κB/NLRP3 signaling pathway.The contents of tumor necrosis factorα(TNF-α),IL-1βand iNOS were detected by ELISA.In addition,the expression levels of IL-6,IL-10,Bax and Bcl-2 proteins in BV2 cells were detected after administration of NF-κB inhibitor BAY 11-7082.The experimental data were analyzed by t test,one-way ANOVA and Dunnett-t test.Results Compared with the control group,the inflammatory cytokines TNF-α(398.80±11.39 vs 113.00±5.93),IL-1β(592.40±11.32 vs 206.70±6.81),IL-6(1.73±0.09 vs 0.99±0.03),iNOS(88.39±2.14 vs 32.48±1.71),p-NF-κB(1.92±0.02 vs 1.02±0.05),INOS(88.39±2.14 vs 32.48±1.71),NLRP3(10.33±0.61 vs 1.02±0.05)and apoptosis-related protein Bax(2.68±0.10 vs 1.05±0.08)in the model group were increased.IL-10(0.82±0.03 vs 1.01±0.06)and Bcl-2 protein expression levels(0.70±0.02 vs 1.11±0.06)were decreased(P<0.05).Western blot results showed that compared with the control group,the protein expression levels of p-NF-κB(2.65±0.06 vs 1.04±0.05)and NLRP3(4.38±0.08 vs 1.07±0.04)in the model group were increased(P<0.001).Compared to the model group,the expression levels of TNF-α(207.90±7.37 vs 398.80±11.39),IL-1β(405.40±6.56 vs 592.40±11.32),IL-6(1.09±0.05 vs 1.73±0.09),iNOS(39.52±1.32 vs 88.39±2.14),p-NF-κB(01.21±0.05 vs 1.92±0.02)and NLRP3(1.51±0.11 vs 10.33±0.61)were decreased,and the expression levels of IL-10(2.61±0.03 vs 0.82±0.03)were increased after treatment with high concentration of curcumin.Compared

关 键 词：姜黄素 NF-κB/ NLRP3信号通路 炎症损伤 细胞凋亡 

分 类 号：R741[医药卫生—神经病学与精神病学]