基于肠道菌群介导的短链脂肪酸调节小胶质细胞极化探讨黄连温胆汤加减方对失眠大鼠的影响  

Modified Coptis Decoction for warming gall bladder regulates microglial polarization through intestinalmicrobiota-mediated SCFAs to improve sleep symptoms in insomnia rats

作　　者：史敏[1] 赵欢[1] 李钊颖 杨东东[1] SHI Min;ZHAO Huan;LI Zhaoying;YANG Dongdong(The Affiliated Hospital of Chengdu University of Traditional Chinese Medicine,Chengdu 610075,Sichuan,China;Chengdu University of Traditional Chinese Medicine,Chengdu 611137,Sichuan,China)

机构地区：[1]成都中医药大学附属医院,四川成都610075 [2]成都中医药大学,四川成都611137

出　　处：《现代中西医结合杂志》2024年第17期2342-2352,共11页Modern Journal of Integrated Traditional Chinese and Western Medicine

基　　金：四川省科技发展计划项目(2023NSFSC1767);成都中医药大学“杏林学者”计划项目(MPRC2022011)。

摘　　要：目的探讨黄连温胆汤加减方对对氯苯丙氨酸(PCPA)诱导的失眠大鼠的影响及其作用机制。方法将50只SD大鼠随机分为正常组、模型组、低剂量黄连温胆汤加减方组、中剂量黄连温胆汤加减方组、高剂量黄连温胆汤加减方组,每组10只。除正常组外,其他组大鼠连续2 d腹腔注射PCPA混悬液350 mg/kg建立失眠模型。从第3天开始,低、中、高剂量黄连温胆汤加减方组分别给予0.845 g/100 g、1.69 g/100 g、3.38 g/100 g的黄连温胆汤加减方灌胃,正常组和模型组给予等量生理盐水灌胃,持续灌胃14 d。对各组大鼠进行Morris水迷宫试验(记录中心区域的移动距离和潜伏时间)、戊巴比妥钠诱导睡眠试验(记录睡眠潜伏期、睡眠持续时间),蛋白印迹法检测各组大鼠海马组织中肿瘤坏死因子-α(TNF-α)、白细胞介素-10(IL-10)、白细胞介素-6(IL-6)、叉头框蛋白P3(FOXP3)表达情况,免疫荧光染色观察各组大鼠海马组织中CD80^(+)/Iba1^(+)和CD206^(+)/Iba1^(+)小胶质细胞表达情况;采用16S rDNA技术检测正常组、模型组、高剂量黄连温胆汤加减方组大鼠肠道菌群多样性及物种组成,进行生物信息学分析及代谢通路预测;采用GC-MS法测定正常组、模型组、高剂量黄连温胆汤加减方组大鼠粪便中各种短链脂肪酸含量;Pearson相关分析模型组大鼠粪便中短链脂肪酸与炎性细胞因子间的关系。结果与正常组比较,模型组大鼠的中心区域移动距离和潜伏时间、睡眠潜伏期均明显延长(P均<0.05),睡眠持续时间明显缩短(P<0.05),海马组织中TNF-α、IL-6蛋白相对表达量及CD80^(+)/Iba1^(+)表达荧光面积均明显升高(P均<0.05),海马组织中IL-10、FOXP3蛋白相对表达量及CD206^(+)/Iba1^(+)表达荧光面积均明显降低(P均<0.05);肠道菌群丰富度明显下降,其中Simpson指数和Shannon指数均明显降低(P均<0.05),肠道菌群相关的代谢通路主要集中在脂质代谢,粪便中乙酸、�Objective It is to investigate the effects of modified Coptis Decoction for warming gall bladder(CDWGB)on p-chlorophenylalanine(PCPA)-induced insomnia in rats and its mechanism of action.Methods Fifty SD rats were randomly divided into normal group,model group,low-dose,medium-dose and high-dose groups of CDWGB,with 10 rats in each group.The rats of all groups except for the normal group were continuously injected intraperitoneally with PCPA suspension 350 mg/kg for 2 days to establish models of insomnia.Starting from the 3rd day,the low-dose,medium-dose and high-dose groups of CDWGB were given modified CDWGB 0.845 g/100 g,1.69 g/100 g,and 3.38 g/100 g by gavage,respectively,and the normal group and model group were given an equal amount of saline by gavage,all continuously gavaged for 14 days.The rats in each group were tested by the Morris water maze(to record the distance traveled in the central area and the latency time)and sodium pentobarbital-induced sleep test(to record the latency and duration of sleep),the expressions of tumor necrosis factor-α(TNF-α),interleukin-10(IL-10),interleukin-6(IL-6)and FOXP3 in hippocampal tissues of rats in each group were detected by protein blotting,the expressions of CD80^(+)/Ibaa in hippocampal tissues of rats in each group were detected by immunofluorescent staining;the expressions of CD80^(+)/Iba1^(+)and CD206^(+)/Iba1^(+)microglia in hippocampus of rats in each group were observed by immunofluorescence staining;the diversity and species composition of the intestinal flora in the normal group,model group and high-dose group of CDWGB were detected by 16S rDNA technology,to analyze the bioinformatics and predict metabolic pathways;the contents of short-chain fatty acids in the feces of rats in the normal group,model group and high-dose group of CDWGB were determined by GC-MS;the relationship between short-chain fatty acids in feces and inflammatory cytokines in the rats of the model group was analyzed by Pearson correlation analysis.Results Compared with the normal group,

关 键 词：黄连温胆汤加减方 失眠 炎症反应 肠道菌群 短链脂肪酸 小胶质细胞 

分 类 号：R-332[医药卫生]