川陈皮素抑制BV2小胶质细胞炎症反应的机制  

Mechanism by which nobiletin inhibits inflammatory response of BV2 microglia

作　　者：迟文鑫 张存鑫 高凯 吕超亮[2] 张科峰[2] Chi Wenxin;Zhang Cunxin;Gao Kai;Lyu Chaoliang;Zhang Kefeng(School of Clinical Medicine,Jining Medical University,Jining 272067,Shandong Province,China;Department of Spine Surgery,Jining No.1 People’s Hospital,Jining 272000,Shandong Province,China;Department of Joint Surgery,Jining No.1 People’s Hospital,Jining 272000,Shandong Province,China)

机构地区：[1]济宁医学院临床医学院,山东省济宁市272000 [2]济宁市第一人民医院脊柱外科,山东省济宁市272000 [3]济宁市第一人民医院关节外科,山东省济宁市272000

出　　处：《中国组织工程研究》2025年第7期1321-1327,共7页Chinese Journal of Tissue Engineering Research

基　　金：山东省自然科学基金(ZR2021LZY008),项目负责人:高凯;济宁市重点研发计划项目(2021YXNS137),项目负责人:高凯;济宁市重点研发计划项目(2021YXNS001),项目负责人:吕超亮。

摘　　要：背景:有研究证实,川陈皮素可改善脂多糖诱导的小胶质细胞异常激活、炎症因子的过度释放和氧化还原失衡,但具体的作用机制尚不充分。目的:探讨川陈皮素抑制脂多糖诱导BV2小胶质细胞炎症反应的分子机制。方法:将第3代BV2小胶质细胞分3组处理:对照组常规培养24 h(不进行任何处理),脂多糖组加入10μg/mL脂多糖处理24 h,脂多糖+川陈皮素组加入20μmol/L川陈皮素处理6 h后加入10μg/mL脂多糖处理24 h。处理结束后,采用CCK-8法检测细胞增殖,活性氧荧光探针检测细胞内活性氧水平,qRT-PCR法检测细胞内核因子κB p65、肿瘤坏死因子α、白细胞介素1βmRNA表达,Western Blot法检测细胞内核因子κB p65、p-核因子κB p65、肿瘤坏死因子α、白细胞介素1β蛋白表达。结果与结论:(1)与对照组比较,脂多糖组细胞增殖活性降低(P<0.001);与脂多糖组比较,脂多糖+川陈皮素组细胞增殖活性升高(P<0.001);(2)与对照组比较,脂多糖组细胞内活性氧水平升高(P<0.001);与脂多糖组比较,脂多糖+川陈皮素组细胞内活性氧水平降低(P<0.01);(3)与对照组比较,脂多糖组细胞内肿瘤坏死因子α、白细胞介素1βmRNA表达升高(P<0.001,P<0.01);与脂多糖组比较,脂多糖+川陈皮素组细胞内肿瘤坏死因子α、白细胞介素1βmRNA表达降低(P<0.01,P<0.05);(4)与对照组比较,脂多糖组细胞内p-核因子κB p65、肿瘤坏死因子α、白细胞介素1β蛋白表达升高(P<0.001);与脂多糖组比较,脂多糖+川陈皮素组细胞内p-核因子κB p65、肿瘤坏死因子α、白细胞介素1β蛋白表达降低(P<0.001);(5)结果表明,川陈皮素可能通过抑制核因子κB信号通路减轻脂多糖诱导的BV2小胶质细胞炎症反应。BACKGROUND:Nobiletin has been found to improve lipopolysaccharide-induced abnormal activation of microglia,excessive release of inflammatory factors and redox imbalance.However,the specific mechanism is not fully understood.OBJECTIVE:To investigate the molecular mechanism by which nobiletin can inhibit lipopolysaccharide-induced inflammation in BV2 microglia.METHODS:Passage 3 BV2 microglia were divided into three groups:control group was cultured for 24 hours(without any treatment).Lipopolysaccharide group was treated with 10μg/mL lipopolysaccharide for 24 hours.Lipopolysaccharide+nobiletin group was treated with 20μmol/L nobiletin for 6 hours and then 10μg/mL lipopolysaccharide for 24 hours.After the processing,cell proliferation was detected by CCK-8 assay.The level of intracellular reactive oxygen species was detected by fluorescent probe.The mRNA expression levels of nuclear factorκB p65,tumor necrosis factorα,and interleukin-1βwere detected by qRT-PCR.The protein expression levels of nuclear factorκB p65,p-nuclear factorκB p65,tumor necrosis factorα,and interleukin-1βwere detected by western blot assay.RESULTS AND CONCLUSION:(1)Compared with the control group,the proliferation activity of lipopolysaccharide group was decreased(P<0.001).Compared with the lipopolysaccharide group,the cell proliferation activity of lipopolysaccharide+nobiletin group was increased(P<0.001).(2)Compared with the control group,the level of intracellular reactive oxygen species was increased in the lipopolysaccharide group(P<0.001).Compared with the lipopolysaccharide group,the level of intracellular reactive oxygen species was decreased in the lipopolysaccharide+nobiletin group(P<0.01).(3)Compared with the control group,the mRNA expression levels of tumor necrosis factorαand interleukin-1βwere increased in the lipopolysaccharide group(P<0.001,P<0.01).Compared with the lipopolysaccharide group,mRNA expression levels of tumor necrosis factorαand interleukin-1βwere decreased in the lipopolysaccharide+nobiletin group(P<0.

关 键 词：川陈皮素 核因子ΚB 小胶质细胞 脂多糖 脊髓损伤 细胞增殖 活性氧 炎性因子 

分 类 号：R459.9[医药卫生—治疗学] R319[医药卫生—临床医学] R681.57