苦参素通过MAPK信号通路对EAE小鼠巨噬细胞M1/M2极化的影响  

Effects of matrine on macrophage M1/M2 polarization in EAE mice through MAPK signaling pathway

作　　者：赵晓玉[1] 朱琳[1] 史香芬[1] 刘楠[1] 楚尧娟[1] 张婉 张英杰[1] 杜书章[1] ZHAO Xiaoyu;ZHU Lin;SHI Xiangfen;LIU Nan;CHU Yaojuan;ZHANG Wan;ZHANG Yingjie;DU Shuzhang(Department of Pharmacy,the First Affiliated Hospital,Zhengzhou University,Zhengzhou 450052)

机构地区：[1]郑州大学第一附属医院药学部,郑州450052

出　　处：《郑州大学学报（医学版）》2022年第1期32-38,共7页Journal of Zhengzhou University(Medical Sciences)

基　　金：国家自然科学基金项目(31870334)。

摘　　要：目的:探讨苦参素通过丝裂原活化蛋白激酶(MAPK)信号通路对实验性自身免疫性脑脊髓炎(EAE)小鼠巨噬细胞M1/M2极化的影响。方法:50只雌性C57BL/6小鼠分为对照组、EAE组、醋酸泼尼松(PA)组、苦参素组及激动剂组,每组各10只,除对照组外均建立EAE模型。于发病日开始给药干预,苦参素组腹腔注射200 mg/kg苦参素注射液,EAE组和对照组腹腔注射10 mL/kg生理盐水,激动剂组腹腔注射200 mg/kg苦参素注射液+2 mg/kg p38MAPK特异性激动剂茴香霉素,PA组灌胃6 mg/kg PA溶液,1次/d,连续给药14 d。取脊髓组织,采用HE染色及LFB染色检测炎性细胞浸润、髓鞘脱失情况;免疫荧光双染法检测M1型、M2型巨噬细胞,qRT-PCR检测iNOS、TNF-α、精氨酸酶-1(Arg-1)、IL-10 mRNA表达情况,Western blot法检测p38MAPK、p-p38MAPK蛋白表达情况。结果:与EAE组比较,PA组、苦参素组及激动剂组脊髓组织炎性细胞浸润评分、髓鞘脱失评分均降低,但仍高于对照组,且PA组<苦参素组<激动剂组(P<0.05)。与EAE组比较,PA组、苦参素组及激动剂组脊髓组织M1型巨噬细胞比例,iNOS、TNF-αmRNA相对表达量及p-p38MAPK/p38MAPK均降低,但仍高于对照组,且PA组<苦参素组<激动剂组(P<0.05);与EAE组比较,PA组、苦参素组及激动剂组脊髓组织M2型巨噬细胞比例及Arg-1、IL-10 mRNA相对表达量均升高,均高于对照组,且PA组>苦参素组>激动剂组(P<0.05)。结论:苦参素可抑制脊髓组织炎症及髓鞘脱失,诱导巨噬细胞向M2型极化,其作用机制可能与抑制p38MAPK磷酸化有关。Aim:To investigate the effects of matrine on macrophage M1/M2 polarization in experimental autoimmune encephalomyelitis(EAE)mice through mitogen-activated protein kinase(MAPK)signaling pathway,and to explore the regulation mechanism.Method:Fifty female C57BL/6 mice were divided into control group,EAE group,prednisolone acetate(PA)group,matrine group and agonist group,with 10 mice in each group.Except for the control group,the EAE model was established and the administration was started on the day of onset.The matrine group was intraperitoneally injected with 200 mg/kg matrine injection,the EAE group and the control group were intraperitoneally injected with 10 mL/kg normal saline,the agonist group was intraperitoneally injected with 200 mg/kg oxymatrine injection+2 mg/kg p38MAPK specific agonist anisomycin,and the PA group was given intragastrically 6 mg/kg PA solution once a day for 14 consecutive days.The spinal cord tissue was taken,HE staining and LFB staining were used to detect inflammatory cell infiltration and demyelination.Macrophage M1/M2 polarization was detected by immunofluorescence double staining.The mRNA expressions of iNOS,TNF-α,Arg-1,IL-10 were detected by qRT-PCR.The protein expressions of p38MAPK and p-p38MAPK were detected by Western blot.Results:Compared with the EAE group,the scores of inflammatory cell infiltration and demyelination of the spinal cord in the PA group,matrine group and agonist group were reduced,but still higher than those of the control group,of which,PA group<matrine group<agonist group(P<0.05).Compared with the EAE group,the proportion of M1 type macrophages in the spinal cord,the relative expressions of iNOS,TNF-αmRNA,and the value of p-p38MAPK/p38MAPK in the PA group,matrine group and agonist group were all decreased,but still higher than those of the control group,of which,PA group<matrine group<agonist group(P<0.05).Compared with the EAE group,the proportion of M2 type macrophages in the spinal cord,the relative expressions of Arg-1 and IL-10 mRNA in the PA group,m

关 键 词：实验性自身免疫性脑脊髓炎 苦参素 巨噬细胞 丝裂原活化蛋白激酶 小鼠 

分 类 号：R744.51[医药卫生—神经病学与精神病学]