四神丸对结肠炎小鼠记忆性T细胞亚群平衡的调控作用  

Regulation of Sishen pills on the balance of memory T cell subsets in colitis mice

作　　者：邓逸菲 黄佳琦 周文 赵海梅[4] 刘端勇[4] 龙健 李燕珍 DENG Yi-fei;HUANG Jia-qi;ZHOU Wen;ZHAO Hai-mei;LIU Duan-yong;LONG Jian;LI Yan-zhen(School of Clinical Medicine,Jiangxi University of Chinese Medicine,Nanchang,330004,Jiangxi,China;Graduate School,Jiangxi University of Chinese Medicine,Nanchang,330004,Jiangxi,China;Scientific research office,Nanchang Medical College,Nanchang,330052,Jiangxi,China;Fangzheng Research Center,Jiangxi University of Chinese Medicine,Nanchang,330004,Jiangxi,China;School of Clinical Medicine,Nanchang Medical College,Nanchang,330052,Jiangxi,China)

机构地区：[1]江西中医药大学临床医学院,江西南昌330004 [2]江西中医药大学研究生院,江西南昌330004 [3]南昌医学院科研处,江西南昌330052 [4]江西中医药大学方证研究中心,江西南昌330004 [5]南昌医学院临床医学院,江西南昌330052

出　　处：《时珍国医国药》2024年第10期2344-2350,共7页Lishizhen Medicine and Materia Medica Research

基　　金：国家自然科学基金项目(82060799);国家级大学生创新训练项目(202310412041);江西省教育厅科学技术项目(GJJ218916);江西中医药大学博士科研启动基金课题(20224BSZR010);井冈学者特聘教授资助计划(1262100102)。

摘　　要：目的探讨四神丸(Sishen pills,SSP)对葡聚糖硫酸钠(Dextran Sulfate Sodium,DSS)诱导的结肠炎小鼠的记忆性T细胞亚群平衡的调控作用及其可能的作用机制。方法40只雄性BALB/c小鼠随机分为4组,分别为:正常组(Control,Ctrl),对照组(Ctrl+SSP),模型组(DSS)和治疗(DSS+SSP)组,每组10只。模型组及治疗组先予以3%DSS(w/v)溶液5d,继而正常饮食饮水7d,再予以2%DSS(w/v)溶液5d。自实验第11天,对照组及治疗组小鼠给予四神丸(2.5g·kg^(-1))灌胃治疗7d。观察小鼠一般生活习性,体质量、结肠质量、结肠质量指数、结肠质量与长度比值等情况;利用HE染色法进行结肠组织病理学观察,并进行病理损伤评分;采用ELISA法检测结肠组织中的抑炎因子IL-2、IL-15、IFN-γ和促炎因子IL-9、IL-12表达水平;流式细胞术检测小鼠脾脏中枢型记忆性T细胞(T_(CM),CD45RA^(-)CD62L^(+)CCR7^(+))和效应型记忆性T细胞(T_(EM),CD45RA^(-)CD62L^(-)CCR7^(-))及其各自CD4/CD8细胞亚群水平。结果与模型组相比,治疗组小鼠炎性细胞浸润程度及溃疡数量明显减少,隐窝修复,腺体排列整齐;在给药治疗后体质量明显增重,结肠长度明显增长,结肠质量、结肠质量指数及病理组织学评分显著下降(P<0.05或P<0.01);小鼠结肠组织中IL-2、IL-15、IFN-γ表达水平明显下降(P<0.05或P<0.01),IL-9、IL-12表达水平明显升高(P<0.05或P<0.01);治疗组T_(CM)细胞水平明显下降(P<0.01),其中CD4^(+)及CD8^(+)T_(CM)细胞水平均显著下降(P<0.05或P<0.01)、TEM、CD4^(+)TEM和CD8^(+)TEM细胞水平均出现明显下降(P<0.01)。结论四神丸能有效治疗小鼠结肠炎,可能与其对记忆性T细胞亚群平衡的调控作用密切相关。Objective To investigate the effect of Sishen pills(SSP)on the balance of memory T cell subsets in mice induced by Dextran Sulfate Sodium(DSS)and its possible mechanism.Methods 40 male BALB/c mice were randomly divided into 4 groups:normal group(Control,Ctrl),control group(Ctrl+SSP),model group(DSS)and treatment group(DSS+SSP),with 10 mice in each group.The model group and the treatment group were first given 3%DSS(w/v)solution for 5 days,followed by normal diet and water for 7 days,and then given 2%DSS(w/v)solution for 5 days.From the 11th day of the experiment,mice in the control group and the treatment group were given Sishen pill(2.5g·kg~(-1))by gavage for 7 days.The general life habits,body mass,colon mass,colon mass index,the ratio of colon weight to colon length were observed.HE staining was used to observe the histopathology changes of the colon,and pathological injury score was performed under the microscope.The expression levels of anti-inflammatory factors IL-2,IL-15,IFN-γand pro-inflammatory factors IL-9 and IL-12 in colon tissues were detected by ELISA.The levels of central memory T cells(T_(CM),CD45RA^(-)CD62L^(+)CCR7^(+))and effector memory T cells(T_(EM),CD45RA^(-)CD62L^(-)CCR7^(-))and their respective CD4/CD8 cell subsets were detected by flow cytometry.Results Compared with the model group,the degree of inflammatory cell infiltration and the number of ulcers in the treatment group were significantly reduced,the colonic crypts were repaired,and glands were arranged neatly.After drug administration,the body mass and colon length were increased significantly,and colon mass,colon mass index and histopathological score were decreased significantly(P<0.05 or P<0.01).The expression levels of IL-2,IL-15 and IFN-γwere significantly decreased(P<0.05 or P<0.01),while the expression levels of IL-9 and IL-12 were significantly increased(P<0.05 or P<0.01).In the treatment group,the levels of T_(CM)cells were significantly decreased(P<0.01),while CD4^(+)and CD8^(+)T_(CM)cells levels were significantly decre

关 键 词：四神丸 溃疡性结肠炎 记忆性T细胞 作用机制 

分 类 号：R285.5[医药卫生—中药学]