苓桂术甘汤抑制脂多糖诱导小胶质细胞由M2向M1型极化作用机制  

Mechanism of Linggui Zhugan decoction on polarization of microglia from M2 to M1 induced by lipopolysaccharide

作　　者：赵小云 李心仪 周春祥[1] ZHAO Xiaoyun;LI Xinyi;ZHOU Chunxiang(Nanjing University of Chinese Medicine,Nanjing 210046,China)

机构地区：[1]南京中医药大学,江苏南京210046

出　　处：《陕西中医》2023年第11期1503-1508,共6页Shaanxi Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金资助项目(82074504,81774021);国家中医药管理局高水平中医药重点学科建设项目[国中医药人教函(2023)85号]。

摘　　要：目的:探讨苓桂术甘汤对阿尔茨海默病(AD)神经炎症细胞模型M2、M1型极化的影响。方法:用浓度为1μg/ml的脂多糖(LPS)刺激小胶质细胞(BV2)24 h,建立细胞神经炎症模型,对细胞进行分组,对照组、模型组、米诺环素组、苓桂术甘汤组。给药结束后,倒置显微镜观察细胞形态变化;硝酸还原酶法测细胞一氧化氮(NO)释放量;ELISA法测细胞上清肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)、白细胞介素-10(IL-10)含量;免疫荧光细胞化学染色法检测M1型标志物COX-2、M2型标志物CD206平均荧光强度。结果:苓桂术甘汤可减少并逆转M2型向M1型转变。模型组NO含量高于对照组,苓桂术甘汤组NO低于模型组,差异有统计学意义(均P<0.05)。模型组TNF-α、IL-1β、IL-6高于对照组,抗炎因子IL-10低于对照组,差异有统计学意义(均P<0.05)。苓桂术甘汤组TNF-α、IL-1β、IL-6高于模型组,IL-10低于模型组,差异有统计学意义(均P<0.05)。模型组COX-2平均荧光强度高于对照组,CD206低于对照组,差异有统计学意义(均P<0.05)。苓桂术甘汤组COX-2低于模型组,CD206高于模型组,差异有统计学意义(均P<0.05)。结论:苓桂术甘汤通过减少TNF-α、IL-1β、IL-6等促炎因子分泌,增加IL-10抗炎因子分泌,减少并逆转小胶质细胞由M2型向M1型极化,发挥抗炎修复作用。Objective:To investigate effect of Linggui Zhugan decoction on M2 and M1 polarization of neuroinflammatory cell model of Alzheimer’s disease(AD).Methods:Microglia(BV2)were stimulated with lipopolysaccharide(LPS)at concentration of 1μg/ml for 24 hours to establish model of cellular neuritis.Grouping the cells,divided into control group,model group,minocycline group and Linggui Zhugan decoction group.After administration,the morphological changes of cells were observed under inverted microscope.The release of nitric oxide(NO)was measured by nitrate reductase method.The contents of tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),interleukin-6(IL-6)and interleukin-10(IL-10)in the supernatant were measured by ELISA method.The average fluorescence intensity of M1 marker COX-2 and M2 marker CD206 were detected by immunofluorescence cytochemical staining.Results:Linggui Zhugan decoction can reduce and reverse the transformation from M2 to M1.The content of NO in the model group was higher than that in the control group,and that in the Linggui Zhugan decoction group lower than that in the model group,difference statistically significant(all P<0.05).TNF-α,IL-1βand IL-6 in the model group were higher than those in the control group,and the anti-inflammatory factor IL-10 lower than that in the control group,difference statistically significant(all P<0.05).TNF-α,IL-1βand IL-6 in the Linggui Zhugan decoction group were higher than those in the model group,and IL-10 lower than that in the model group,difference statistically significant(all P<0.05).The average fluorescence intensity of COX-2 in the model group was higher than that in the control group,and CD206 lower than that in the control group,difference statistically significant(all P<0.05).COX-2 in the Linggui Zhugan decoction group was lower than that in the model group,CD206 higher than that in the model group,difference statistically significant(all P<0.05).Conclusion:Linggui Zhugan decoction may reduce the secretion of pro-inflammatory factors such as

关 键 词：阿尔茨海默病 神经炎症 苓桂术甘汤 BV2小胶质细胞 脂多糖 痰饮证 

分 类 号：R74[医药卫生—神经病学与精神病学]