二甲双胍抑制创伤性脑损伤小鼠炎症的机制  

Mechanism of metformin in inhibiting inflammation of mice with traumatic brain injury

作　　者：王钊 包海军 孙卓 蔡红星 WANG Zhao;BAO Haijun;SUN Zhuo(School of Forensic and Pathology,Xuzhou Medical University,Xuzhou 221002,CHINA)

机构地区：[1]徐州医科大学法医学教研室,江苏221002 [2]徐州医科大学临床与实验病理学实验室,江苏221002 [3]徐州医科大学江苏省医学基因检测工程中心

出　　处：《江苏医药》2020年第4期337-341,F0003,共6页Jiangsu Medical Journal

基　　金：江苏省重点实验室开放课题(JBSL1703);江苏省普通高校研究生科研、实践创新计划(KYCX19-2223)。

摘　　要：目的探讨二甲双胍抑制创伤性脑损伤(TBI)小鼠炎症的机制。方法实验分为C57BL/6小鼠野生型组、野生型小鼠生理盐水组、野生型小鼠二甲双胍组、FoxO3A基因敲除组及基因敲除小鼠二甲双胍组。二甲双胍给药方式为每日采用二甲双胍150 mg/kg灌胃,连续7 d。维持给药7 d后,建立小鼠6、12、24 h TBI模型。应用Western blot法检测FoxO3A蛋白、LC3、P62以及炎症因子IL-1β、NLRP3的蛋白表达。免疫荧光双染法检测LC3、IL-1β的表达情况。碘化丙啶染色计数神经细胞坏死数量。结果FoxO3A在TBI后促进自噬的发生,同时抑制炎症。二甲双胍通过抑制FoxO3A的磷酸化,增强FoxO3A的表达,促进自噬的发生进而抑制炎症,并减少神经细胞的坏死数量。结论TBI后,FoxO3A参与自噬的发生;二甲双胍通过FoxO3A增强自噬、抑制炎症。Objective To investigate the mechanism of metformin in inhibiting inflammation of mice with traumatic brain injury.Methods The experiment mice were divided into wild-type C57 BL/6 mice group(group A),wild-type mice with saline administration group(group B),wild-type mice with metformin administration group(group C),FoxO3 A knockout mice group(group D)and FoxO3 A knockout mice with metformin administration group(group E).The mice in groups of C and E were administered daily with metformin 150 mg/kg by gastric gavage for 7 consecutive days.The 6,12 and 24 h models of brain trauma were established in five mice in each group.Western blot was used to detect the expression of FoxO3 A protein,LC3,P62,IL-1βand NLRP3.Meanwhile,LC3 and IL-1βwere detected by immunofluorescence double staining.PI staining was used to count the number of nerve cell necrosis.Results FoxO3 A promoted autophagy and inhibited inflammation in the mice after traumatic brain injury.By inhibiting the phosphorylation of FoxO3 A,metformin inhibited phosphorylated-FoxO3 A,enhanced FoxO3 A expression,inhibited inflammation and reduced the number of dead nerve cells.Conclusion FoxO3 A is involved in autophagy in the mice after traumatic brain injury.Metformin enhances autophagy and inhibits inflammation through FoxO3 A.

关 键 词：创伤性脑损伤 二甲双胍 FOXO3A 自噬 炎症 

分 类 号：R392[医药卫生—免疫学]