醒脑解郁方含药血清对IL-4+LPS诱导的小胶质细胞极化、细胞活力的影响  

Effects of Xingnao Jieyu decoction containing serum on IL-4+LPS-induced polarization and cell viability of microglia

作　　者：谢欣纯 杨一帆 彭涛 闫咏梅[1,2] XIE Xinchun;YANG Yifan;PENG Tao;YAN Yongmei(The First Clinical Medical School,Shanxi University of Traditional Chinese Medicine,Xianyang 712046,China;不详)

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]陕西中医药大学附属医院脑病医院

出　　处：《山东医药》2023年第20期11-15,共5页Shandong Medical Journal

基　　金：国家自然科学基金面上项目(81973843);2021年岐黄学者支持项目。

摘　　要：目的 观察醒脑解郁方含药血清对白细胞介素(IL)-4+脂多糖(LPS)诱导的小胶质细胞M1/M2型极化、细胞活力及炎症因子水平的影响,以探讨醒脑解郁方治疗卒中后抑郁的机制。方法 将SD大鼠随机分为醒脑解郁方低、中、高剂量组和对照组,分别予0.525、1.05、2.1 g/mL的醒脑解郁方和蒸馏水灌胃,3 d后腹主动脉取血,获得血清。培养BV2细胞,分为空白组、模型组、实验1组、实验2组、实验3组;实验1、2、3组及模型组细胞先后接种于含20 ng/mL IL-4、100μg/mL LPS的胎牛血清培养基各培养12 h,实验1、2、3组分别更换为含低、中、高剂量醒脑解郁方含药血清的胎牛血清培养基,模型组更换为胎牛血清培养基;空白组胎牛血清培养基不添加药物。倒置显微镜下观察各组细胞形态变化,检测细胞上清液中的M1极化相关因子[肿瘤坏死因子α(TNF-α)、IL-6、IL-1β、一氧化氮(NO)]和M2极化相关因子[IL-10、脑源性神经营养因子(BDNF)],采用MTT比色法检测细胞活力。结果 空白组细胞为圆形或椭圆形,基本无突触;IL-4处理后的模型组细胞出现两头分支,胞体未见明显增大,为M2型极化状态;IL-4+LPS处理后的模型组细胞分支明显增多、触角增加,突起变短变粗,胞体增大,呈“阿米巴样”,为M1型极化状态;实验1、2、3组“阿米巴样”细胞减少,短粗突触变长,胞体变小或变椭圆,恢复M2型极化状态。模型组细胞上清液中NO、IL-1β、IL-6、TNF-α水平高于空白组,IL-10、BDNF低于空白组(P均<0.05);实验1、2、3组的NO、IL-1β、IL-6、TNF-α水平低于对照组,IL-10、BDNF水平高于对照组(P均<0.05)。模型组培养各时点细胞活力均低于空白组(P均<0.05);培养24 h时,实验1、2、3组细胞活力高于模型组,以实验2组细胞活力最高(P均<0.05)。结论 醒脑解郁方含药血清可抑制M2型小胶质细胞向M1型转化,发挥抗炎作用及神经保护作用,从而起到治疗卒中后�Objective To observe the effects of Xingnao Jieyu decoction containing serum on M1/M2 polarization,cell viability,and inflammatory factor levels of microglia induced by interleukin(IL)-4 and lipopolysaccharide(LPS),in order to explore the mechanism of Xingnao Jieyu decoction containing serum in the treatment of post-stroke depression.Methods SD rats were randomly divided into the low-dose,medium-dose,high-dose Xingnao Jieyu decoction groups,and control group,which were treated with 0.525,1.05,2.1 g/mL Xingnao Jieyu decoction,and distilled water,respectively,for three days.Blood samples were collected from the abdominal aorta to obtain serum.BV2 cells were cultured and divided into the blank group,model group,and experimental groups 1,2,and 3,respectively.Cells in the experimental groups and model group were treated with IL-4(20 ng/mL) and LPS(100 μg/mL) in fetal bovine serum medium for 12 h.Then,cells in the experimental groups were further treated with low-dose,medium-dose,and high-dose Xingnao Jieyu decoction containing serum,while cells in the model group were treated with fetal bovine serum medium;cells in the blank group were cultured in the fetal bovine serum medium without any drug.Changes in cell morphology were observed under the inverted microscope,and M1 polarization-related factors [tumor necrosis factor-α(TNF-α),interleukin(IL)-6,IL-1β,and nitric oxide(NO)] and M2 polarization-related factors [IL-10 and brain-derived neurotrophic factor(BDNF)] in cell supernatant were measured.Cell viability was assessed using MTT colorimetry.Results Cells in the blank group were round or elliptical with almost no synapses.Cells in the model group treated with IL-4 showed two branches,no obvious enlargement of the cell body,indicating M2 polarization.Cells in the model group treated with IL-4+LPS showed increased branches and protrusions,shorter and thicker processes,and enlarged cell bodies with an "amoeboid" appearance,indicating M1 polarization.Cells in the experimental groups 1,2,and 3 showed reduced "amoeboi

关 键 词：醒脑解郁方 卒中后抑郁 脑卒中 小胶质细胞 炎症反应 

分 类 号：R741.05[医药卫生—神经病学与精神病学]