基于TLR4/NF-κB信号通路探讨逍遥散改善IBS-D小鼠肠道炎症及抑郁状态的机制研究OA
Mechanism of Xiaoyao San in Alleviating Intestinal Inflammation and Depressive-like Behavior in IBS-D Mice:A Study Based on the TLR4/NF-κB Signaling Pathway
目的:从肠道低炎症角度出发,探究逍遥散对慢性不可预知应激(CUMS)诱导的腹泻型肠易激综合征(IBS-D)小鼠肠道炎症及抑郁状态的改善作用与分子机制.方法:将60只健康SPF级小鼠随机分为空白组、模型组、逍遥散低(XYS-L)、中(XYS-M)、高(XYS-H)剂量组,每组12只.除空白组外,其余各组通过连续4周采用CUMS刺激联合7天法番泻叶灌胃(0.5 g/mL,0.1 mL/10g)建立肝郁脾虚型IBS-D小鼠模型.造模成功后,空白组与模型组每日灌胃生理盐水,逍遥散组各组分别灌胃0.59、1.18、2.36 g/mL的逍遥散水煎液,每天给药1次,连续干预7天.观察小鼠体重、粪便、精神状态等一般情况,检测腹泻率、粪便含水率及糖水偏嗜率;HE染色观察结肠组织病理学变化;RT-qPCR、Western blot法检测结肠组织中TLR4/NF-κB通路相关mRNA及蛋白表达;ELISA法测定血清白细胞介素-6(IL-6)、白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)水平.结果:逍遥散干预后,与模型组小鼠比较,逍遥散各剂量组小鼠体重显著增加,粪便含水率降低,糖水偏嗜率升高,摄食量恢复(P<0.05);HE染色显示,模型组小鼠结肠黏膜上皮呈绒毛状或指状突起,黏膜上层炎细胞浸润明显,逍遥散各剂量组黏膜病变均得到改善,其中XYS-M组小鼠黏膜上皮平整,绒毛状或指状结构消失,炎细胞浸润程度显著减轻;同时,XYS-H组与模型组比较,XYS-H组可显著下调结肠组织TLR4 mRNA及蛋白、NF-κB p65蛋白的表达(P<0.05),上调IkBα蛋白表达(P<0.05),并降低血清IL-1β、IL-6、TNF-α浓度(P<0.05).结论:逍遥散可能通过抑制TLR4/NF-κB通路,减少促炎因子的释放,进而改善IBS-D小鼠肠道低度炎症与抑郁状态,为中医药治疗IBS-D合并情绪障碍提供实验依据.
Objective:To investigate the therapeutic effects and underlying molecular mechanisms of Xiaoyao San(XYS)on intestinal low-grade inflammation and depressive-like behavior in mice with Chronic Unpredictable Mild Stress(CUMS)-induced Diarrhea-predominant Irritable Bowel Syndrome(IBS-D).Methods:Sixty SPF mice were randomly assigned to five groups(n=12):Control,Model,XYS low-dose(XYS-L,0.59 g/mL),XYS medium-dose(XYS-M,1.18 g/mL),and XYS high-dose(XYS-H,2.36 g/mL).The IBS-D model with liver depression and spleen deficiency pattern was established in all groups except the Control by 4 weeks of CUMS combined with 7-day senna decoction gavage(0.5 g/mL,0.1 mL/10g).Post-modeling,the Control and Model groups received normal saline,while the XYS groups received their respective XYS decoctions for 7 days.General status(body weight,fecal morphology,mental state),diarrhea rate,fecal water content,and sucrose preference were assessed.Colon histopathology was examined via HE staining.mRNA and protein expression of TLR4 and NF-κB in colon tissue were measured by RT-qPCR and Western blot.Serum levels of IL-6,IL-1β,and TNF-α were quantified by ELISA.Results:Compared with the Model group,XYS intervention significantly improved general status,reduced fecal water content,and increased sucrose preference.HE staining showed that the Model group had villous/finger-like mucosal protrusions and marked inflammatory infiltration,which were alleviated in the XYS-M group.Meanwhile,compared with the model group,the XYS-H group significantly downregulated the expression of TLR4 mRNA and protein,as well as NF-κB p65 protein in the colon tissue(P<0.05),upregulated the expression of IkBα protein(P<0.05),and decreased the concentrations of serum IL-1β,IL-6,and TNF-α(P<0.05).Conclusion:Xiaoyao San may alleviate intestinal low-grade inflammation and depressive-like behavior in IBS-D mice,potentially by inhibiting the TLR4/NF-κB pathway and subsequent pro-inflammatory cytokine release.This provides experimental support for the use of traditional Chinese medicine in managing IBS-D with comorbid emotional disorders.
柯晓聪;宋佳;邱子芸;李玲;陈宝国;叶菁
江西中医药大学,江西 南昌 330004江西中医药大学,江西 南昌 330004江西中医药大学,江西 南昌 330004江西中医药大学,江西 南昌 330004江西中医药大学,江西 南昌 330004||江西中医药大学附属医院,江西 南昌 330006江西中医药大学,江西 南昌 330004||江西中医药大学附属医院,江西 南昌 330006
医药卫生
逍遥散腹泻型肠易激综合征抑郁状态TLR4/NF-κB信号通路肠道炎症促炎因子
Xiaoyao Sandiarrhea-predominant irritable bowel syndromedepressive-like behaviorTLR4/NF-κB signaling pathwayintestinal inflammationpro-inflammatory cytokines
《中医康复》 2026 (7)
57-66,10
江西中医药大学重点学科资助项目(2023jzzdxk007)江西省名中医传承工作室建设项目-陈宝国省名中医工作室(赣中医药科教字[2024]8号)
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