摘要
目的探究栀子豉汤的抗抑郁效果和可能作用机制。方法取50只C57BL/6J小鼠随机分为空白对照组、模型组、氟西汀组和栀子豉汤高、低剂量组,每组10只。除空白对照组外,其余各组小鼠采用慢性轻度不可预测应激建立抑郁模型,造模后栀子豉汤低、高剂量组每日分别给予栀子豉汤3.25、6.50 g/kg灌胃,氟西汀组给予氟西汀混悬液20 mg/kg灌胃,空白对照组和模型组每天给予0.1 ml/10 g生理盐水灌胃,各组均灌胃3周。末次给药1 h后依次进行蔗糖偏好测试、悬尾实验、强迫游泳实验和旷场实验检测小鼠抑郁样表现,蛋白免疫印迹法检测小鼠海马中核苷酸结合寡聚化结构域样受体蛋白3(NLRP3)、凋亡相关斑点样蛋白(ASC)、半胱氨酸天冬氨酸蛋白酶1(Caspase-1)、剪切型Gasdermin家族蛋白D(Cleaved GSDMD)、白细胞介素1β(IL-1β)、白细胞介素18(IL-18)以及突触前膜蛋白神经突触素1(Synapsin-1)、突触后致密蛋白95(PSD-95)蛋白表达水平,采用免疫荧光法标记并定量小鼠脑组织离子钙结合衔接分子1(Iba-1)水平,采用高尔基染色法可视化海马神经元的树突与树突棘密度。结果与空白对照组比较,模型组小鼠蔗糖偏好率降低,悬尾实验静止时间和强迫游泳实验静止时间延长,海马区NLRP3、Caspase-1、IL-1β、IL-18、Cleaved GSDMD蛋白表达及脑组织Iba-1蛋白荧光强度升高,Synapsin-1、PSD-95蛋白水平及树突棘密度均明显降低(P<0.05或P<0.01);与模型组比较,栀子豉汤高剂量组上述各指标均明显改善,且栀子豉汤高剂量组在降低IL-1β,升高Synapsin-1、PSD-95蛋白表达方面优于栀子豉汤低剂量组(P<0.05或P<0.01)。结论栀子豉汤对抑郁小鼠行为有改善效果,可能是通过抑制海马NLRP3活化,减少IL-1β、IL-18的释放,改善了突触损伤,增强神经可塑性,从而发挥抗抑郁作用。
ObjectiveTo explore the effect of Zhizichi Decoction(栀子豉汤)on depression and the possiblemechanism.MethodsFifty C57 BL/6 J mice were randomly divided into blank control group,model group,fluox-etine group and Zhizichi Decoction high-and low-dose groups,with 10 mice in each group.Except for the blank con-trol group,the mice were given unpredictable chronic mild stress to induce a depression model.After modeling,thelow-and high-dose Zhizichi Decoction groups were given 3.25 and 6.50 g/kg of Zhizichi Decoction by gavage,respec-tively,while the fluoxetine group was given 20 mg/kg of fluoxetine suspension,and the blank control group was given0.1 ml/10 g of normal saline by gavage,once daily for 3 weeks.One hour after the last medication,the sucrose prefer-ence test,the tail suspension test(TST),the forced swim test(FST)and the open field test were performed in se-quence to detect the depression-like behavior in mice,and the western blotting was used to detect the expression of nu-cleotide-binding oligomerization domain-like receptor 3(NLRP3),apoptosis-associated speck-like protein(ASC),Caspase-1,cleaved Gasdermin D(Cleaved GSDMD),interleukin 1β(IL-1β),interleukin 18(IL-18)in the hippo-campus,as well as the presynaptic membrane protein synaptophysin-1(Synapsin-1)and postsynaptic compact pro-tein 95(PSD-95).Immunofluorescence was used to label and quantify the level of ionized calcium-binding adaptor 1(Iba-1)in the brain tissue,and Golgi staining was used to visualize the density of dendrites and dendritic spines ofhippocampal neurons.ResultsCompared to the blank control group,the model group had decreased sucrose prefer-ence rate,prolonged immobility time in the TST and FST,increased expression of NLRP3,Caspase-1,IL-1β,IL-18,Cleaved GSDMD in hippocampus,increased fluorescence intensity of Iba-1 in brain tissue,decreased Synapsin-1 and PSD-95 levels and dendritic spine density(P<0.05 or P<0.01).The high-dose Zhizichi Decoction group wasbetter than the model group in all the above mentioned outcomes,and showed better effect than low-dose group in re-ducing IL-1βand increasing the expressions of Synapsin-1 and PSD-95(P<0.05 or P<0.01).Conclusion Zhizichi Decoction can improve the behavior of depressed mice,possibly by inhibiting the activation of NLRP3 in thehippocampus,reducing the release of IL-1βand IL-18,improving synaptic damage and enhancing neuroplasticity,thereby exerting an antidepressant effect.
作者
陶伟伟
白子君
岳启予
姚广大
胡玥
陈刚
TAO Weiwei;BAI Zijun;YUE Qiyu;YAO Guangda;HU Yue;CHEN Gang(School of Traditional Chinese Medicine/School of Integrated Traditional Chinese and Western Medicine,Nanjing University of Chinese Medicine,Nanjing,210023;School of Traditional Chinese Medicine,Jinan University)
出处
《中医杂志》
CSCD
北大核心
2022年第11期1073-1079,共7页
Journal of Traditional Chinese Medicine
基金
国家自然科学基金(81873096)
虞山医派中医经典传承奖励基金项目。
关键词
抑郁症
栀子豉汤
脑组织
核苷酸结合寡聚化结构域样受体蛋白3
炎症小体
小胶质细胞
depression
Zhizichi Decoction(栀子豉汤)
brain tissue
nucleotide-binding oligomerization domain-like receptor protein 3
inflammasome
microglia
