洗心汤含药血清对Aβ_(25-35)诱导的BV-2小胶质细胞极化的调控作用  

Xixintang-medicated Serum Regulates Aβ_(25-35)-induced Polarization of BV-2 Microglial Cells

作　　者：杨朝凯 第五永长[2] 武阳洋 邢霞 王登坤 YANG Chaokai;DIWU Yongchang;WU Yangyang;XING Xia;WANG Dengkun(The First Clinical Medical College of Shaanxi University of Chinese Medicine,Xianyang 712046,China;Basic Medical College of Shaanxi University of Chinese Medicine,Xianyang 712046,China;Affiliated Hospital of Shaanxi University of Chinese Medicine,Xianyang 712000,China)

机构地区：[1]陕西中医药大学第一临床医学院,陕西咸阳712046 [2]陕西中医药大学基础医学院,陕西咸阳712046 [3]陕西中医药大学附属医院,陕西咸阳712000

出　　处：《中国实验方剂学杂志》2025年第6期18-26,共9页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金面上项目(82074503);陕西省“特支计划”科技创新领军人才项目(陕组通字[2018]33号);陕西中医药大学神经退行性疾病中医药防治研究学科创新团队项目(2019-QN05);陕西省中医药管理局脑髓病防治重点研究室建设项目;陕西中医药大学高水平中医药重点学科建设项目。

摘　　要：目的:通过体外研究,探索洗心汤(XXT)含药血清对β淀粉样蛋白(Aβ)25-35诱导的BV-2小胶质细胞极化的影响,以揭示该方在防治阿尔茨海默病中的潜在机制,为临床治疗提供科学依据。方法:将BV-2小胶质细胞传代培养后,使用细胞增殖和活性检测(CCK-8)法筛选出最适浓度的XXT含药血清和Aβ_(25-35)。实验设置空白组、模型组(浓度为40μmol·L^(-1)的Aβ_(25-35))、XXT含药血清组(浓度为40μmol·L^(-1)的Aβ_(25-35)+10%XXT含药血清)和空白血清组(浓度为40μmol·L^(-1)的Aβ_(25-35)+10%空白血清处理组)。各组细胞孵育24 h后,采用免疫荧光法检测离子化钙结合适配分子1(IBA1)、CD16/32及CD206的表达;采用实时荧光定量聚合酶链式反应(Real-time PCR)检测CD206、CD163、诱导型一氧化氮合成(iNOS)、精氨酸酶-1(Arg-1)mRNA表达水平;通过酶联免疫吸附测定法(ELISA)检测NGF、iNOS、Arg-1的含量。采用硝酸还原酶法测定NO含量。结果:与空白组比较,模型组IBA1、CD16/32表达显著升高(P<0.01),CD206的表达明显降低(P<0.05),iNOS mRNA表达水平显著上调(P<0.01),含量明显增高(P<0.05),CD206、CD163、Arg-1的mRNA表达水平明显下调(P<0.05,P<0.01),Arg-1、NGF含量明显降低(P<0.05),NO含量明显升高(P<0.05)。与模型组比较,XXT含药血清组IBA1,CD16/32表达明显降低(P<0.05,P<0.01),CD206的表达显著升高(P<0.01);iNOS含量和mRNA表达水平明显下调(P<0.05,P<0.01),同时CD206、CD163、Arg-1的mRNA表达水平显著上调(P<0.01),Arg-1、NGF含量明显升高(P<0.05),NO含量明显降低(P<0.05)。与模型组比较,空白血清组各指标差异无统计学意义。结论:XXT含药血清可通过抑制M1型小胶质细胞极化并促进M2型极化,发挥抗炎和神经营养作用。Objective:To investigate the effects of Xixintang(XXT)-medicated serum on the amyloid β-protein(Aβ)25-35-induced polarization of BV-2 microglial cells by a cell experiment and uncover the potential mechanisms of this formula in the prevention and treatment of Alzheimer's disease(AD),thus providing scientific evidence for the clinical application of XXT.Methods:BV-2 microglial cells were subcultured.The optimal concentrations of XXT-medicated serum and Aβ_(25-35)were determined via the cell-counting kit-8(CCK-8)assay.The cell experiment was carried out with the following groups:blank control,model(Aβ_(25-35)at 40μmol·L^(-1)),XXT-medicated serum(Aβ_(25-35)at 40μmol·L^(-1)+10%XXT-medicated serum),and blank serum(Aβ_(25-35)at 40μmol·L^(-1)+10% blank serum).After 24 hours of cell incubation,immunofluorescence was used to detect the expression of ionized calcium-binding adaptor molecule 1(IBA1),CD16/32,and CD206.Real-time PCR was performed to measure the mRNA levels of CD206,CD163,inducible nitric oxide synthase(iNOS),and arginase 1(Arg-1).Enzyme-linked immunosorbent assay(ELISA)was employed to quantify the levels of nerve growth factor(NGF),iNOS,and Arg-1.The nitric oxide(NO)concentration was determined via the nitrate reductase method.Results:Compared with the blank control group,the model group showed increased expression of IBA1 and CD16/32(P<0.01),decreased expression of CD206(P<0.05),upregulation in the mRNA level(P<0.01)and content(P<0.05)of iNOS,downregulation in the mRNA levels of CD206,CD163,and Arg-1(P<0.05,P<0.01),lowered levels of Arg-1 and NGF(P<0.05),and an elevation in the NO level(P<0.05).Compared with the model group,the XXT-medicated serum group exhibited reduced expression of IBA1 and CD16/32(P<0.05,P<0.01)and increased expression of CD206(P<0.01).Both the content and mRNA level of iNOS were downregulated(P<0.05,P<0.01),while the mRNA levels of CD206,CD163,and Arg-1 were upregulated(P<0.01)in the XXT-medicated serum group.In addition,the XXT-medicated serum group showed elevated levels o

关 键 词：洗心汤 β淀粉样蛋白(Aβ)25-35 BV-2小胶质细胞 细胞极化 神经炎症 

分 类 号：R289[医药卫生—方剂学] R285[医药卫生—中药学] R259[医药卫生—中医学]