益气化痰方抑制NLRP3炎症小体对抑郁小鼠突触功能障碍的调控作用研究  

Effect of Yiqi Huatan decoction in inhibiting NLRP3 inflammasome on the modulation of synaptic dysfunction in depressed mice

作　　者：李咪 董健健 徐银[1] 程楠 韩咏竹 LI Mi;DONG Jianjian;XU Yin;CHENG Nan;HAN Yongzhu(Institute of Neurology of Anhui University of Chinese Medicine,Hefei 230038,China;Center for Xin'an Medicine and Modernization of Traditional Chinese Medicine of IHM,Anhui University of Chinese Medicine,Hefei 230012,China)

机构地区：[1]安徽中医药大学神经病学研究所,安徽合肥230038 [2]安徽中医药大学新安医学与中医药现代化研究所,安徽合肥230012

出　　处：《海南医学院学报》2024年第11期833-842,共10页Journal of Hainan Medical University

基　　金：安徽省高校自然科学研究项目重点项目(KJ2021A0551);安徽中医药大学自然科学研究项目(2020sjzd05)。

摘　　要：目的:研究益气化痰方通过抑制小胶质细胞NLRP3炎症小体和A1型星形胶质细胞激活对慢性温和不可预测应激(CUMS)抑郁小鼠模型突触功能障碍的调控作用。方法:将C57BL/6J小鼠分为正常组、模型组、MCC950组和益气化痰方组。除正常组外,其余各组均采用CUMS建立抑郁模型,分别给予MCC950或益气化痰方进行治疗。通过蔗糖偏好实验、新奇抑制摄食实验、悬尾实验、强迫游泳实验评估小鼠抑郁样行为;高尔基染色法观察小鼠海马突触结构;免疫荧光染色检测小鼠海马GFAP与C3、Iba1与NLRP3的共定位情况;实时荧光定量PCR检测TNF-α、IL-1α、C1qA mRNA表达水平;免疫印迹法检测小鼠海马SYP、PSD-95、C3、NLRP3、ASC、Caspase-1、IL-1β的蛋白表达水平。结果:与正常组比较,模型组小鼠蔗糖偏好率显著降低,新奇抑制摄食实验中摄食潜伏期延长,悬尾实验和强迫游泳实验不动时间延长(P<0.05),海马树突分支数目及树突棘密度减少(P<0.01),GFAP与C3、Iba1与NLRP3共定位阳性细胞数显著增多(P<0.01),TNF-α、IL-1α、C1qA mRNA表达水平显著增高(P<0.01),SYP、PSD-95蛋白表达水平降低(P<0.05),C3、NLRP3、ASC、Caspase-1、IL-1β蛋白表达均显著增高(P<0.05);与模型组比较,益气化痰方组小鼠蔗糖偏好率升高,新奇抑制摄食实验摄食潜伏期、悬尾实验和强迫游泳实验不动时间显著缩短(P<0.01),海马树突分支数目及树突棘密度显著升高(P<0.01),GFAP与C3、Iba1与NLRP3共定位阳性细胞数显著减少(P<0.01),TNF-α、IL-1α、C1qA mRNA水平显著降低(P<0.01),SYP、PSD-95蛋白表达水平升高(P<0.05),C3、NLRP3、ASC、Caspase-1、IL-1β蛋白表达均显著减少(P<0.05)。结论:益气化痰方可通过抑制海马小胶质细胞NLRP3炎症小体活化,减少具有神经毒性的A1型星形胶质细胞激活,从而减轻突触功能损伤,改善CUMS小鼠抑郁样行为。Objective:To explore how Yiqi Huatan Decoction can modulate synaptic dysfunction in a mouse model of chronic unpredictable mild stress(CUMS)depression by suppressing NLRP3 inflammasome in microglia and A1-like astrocytes activa-tion.Methods:C57BL/6J mice were categorized into four groups:normal,model,MCC950,and Yiqi Huatan Decoction.With the exception of the normal group,all other groups underwent CUMS to induce depression,and subsequently received either MCC950 or Yiqi Huatan Decoction as treatment.The depression-like behavior of mice was evaluated using the Sucrose preference test(SPT),Novelty suppressed feeding test(NSF),Tail suspension test(TST),and Forced swimming test(FST).The syn-aptic structure of the hippocampus was examined through Golgi staining.Immunofluorescence staining was utilized to detect the co-localization of GFAP and C3,as well as Iba1 and NLRP3 in the hippocampus of mice.Additionally,the mRNA levels of TNF-α,IL-1α,and C1qA were measured using quantitative Real-time PCR.The protein expression of SYP,PSD-95,C3,NL-RP3,ASC,Caspase-1,and IL-1βin the mouse hippocampus were determined through Western Blot analysis.Results:Com-pared to the control group,the mice in the experimental group exhibited a notable decrease in sucrose preference,an extended peri-od of time before initiating feeding in NSF,and an increased duration of immobility in both TST and FST(P<0.05).In addition,mice exhibiting depressive symptoms demonstrated a reduction in the quantity of dendritic branches and dendritic spine density within the hippocampus(P<0.01).Furthermore,there was a notable rise in the number of co-localized positive cells expressing GFAP and C3 or Iba1 and NLRP3(P<0.01),along with elevated levels of TNF-α,IL-1α,and C1qA mRNA expression(P<0.01).Conversely,a decrease in the expression of SYP and PSD-95 proteins was observed(P<0.05),while a significant increase in the expression of C3,NLRP3,ASC,Caspase-1,and IL-1βproteins was noted(P<0.01).In contrast to the model group,the mice in the Yiqi Huatan Decoction

关 键 词：抑郁症 益气化痰方 NLRP3炎症小体 小胶质细胞 星形胶质细胞 突触功能障碍 

分 类 号：R285.5[医药卫生—中药学]