首页|萸连丸对葡聚糖硫酸钠诱导的溃疡性结肠炎小鼠记忆性滤泡辅助性T细胞的调节作用

萸连丸对葡聚糖硫酸钠诱导的溃疡性结肠炎小鼠记忆性滤泡辅助性T细胞的调节作用

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目的 探究萸连丸(黄连、吴茱萸)对葡聚糖硫酸钠(DSS)诱导的溃疡性结肠炎小鼠脾脏记忆性滤泡辅助性T细胞(Memory follicular T helper cell,mTfh)的调节作用。方法 将 40 只BALB/c小鼠随机分为正常组、模型组、萸连丸组(0。5 g·kg-1)和美沙拉嗪组(0。3 g·kg-1),每组 10 只。采用3%DSS溶液自由饮用 7d诱导溃疡性结肠炎小鼠模型。实验过程每天记录小鼠的精神状态、粪便性状、便血及体质量情况;测量小鼠结肠长度、结肠质量,计算结肠质量指数;采用HE染色法观察结肠组织病理形态变化;ELISA法测定结肠组织中白细胞介素 6(IL-6)、IL-15 的表达水平;流式细胞术测定脾脏组织中mTfh细胞亚群的表达情况;Western Blot法测定结肠组织中Roquin-1、AMPK-α、p-AMPK-α蛋白表达水平。结果 与正常组比较,模型组小鼠的体质量显著下降(P<0。01),结肠长度显著缩短(P<0。01),结肠质量明显增加(P<0。05),结肠质量指数显著升高(P<0。01),结肠组织病理损伤严重;结肠组织中的促炎细胞因子IL-6、IL-15 表达水平显著升高(P<0。01);脾脏组织中CD4+CCR7-CXCR5+CD62L+、CD4+CCR7+CXCR5+GL7+、CD4+CCR7-CXCR5+GL7+细胞表达水平显著升高(P<0。01),而 CD4+CCR7+CXCR5+CD62L+细胞表达水平显著降低(P<0。01);结肠组织中 Roquin-1、AMPK-α、p-AMPK-α蛋白表达水平明显降低(P<0。05)。与模型组比较,萸连丸组及美沙拉嗪组小鼠的体质量明显上升(P<0。05),结肠长度显著延长(P<0。01),结肠质量明显降低(P<0。05),结肠质量指数显著下降(P<0。01),结肠组织病理损伤得到较明显改善;结肠组织中的IL-6、IL-15 表达水平显著降低(P<0。01);脾脏组织中CD4+CCR7-CXCR5+CD62L+、CD4+CCR7+CXCR5+GL7+、CD4+CCR7-CXCR5+GL7+细胞表达水平显著降低(P<0。01),而 CD4+CCR7+CXCR5+CD62L+细胞表达水平显著升高(P<0。01);结肠组织中 Roquin-1、AMPK-α、p-AMPK-α蛋白表达水平明显升高(P<0。05,P<0。01)。结论 萸连丸对DSS诱导的小鼠溃疡性结肠炎具有治疗作用,能够改善结肠组织病理损伤,可能与其激活Roquin-1/AMPK-α信号通路,下调炎性细胞因子IL-6、IL-15 表达,调控mTfh细胞亚群稳态有关。
The Modulatory Effect of Yulian Pills on Memory Follicular Helper T Cells in Mice with Dextran Sulfate Sodium-Induced Ulcerative Colitis
Objective To investigate the modulatory effect of Yulian Pills(composed of Coptidis Rhizoma and Euodiae Fructus)on splenic memory follicular T helper cell(mTfh)in mice with dextran sodium sulfate(DSS)-induced ulcerative colitis.Methods Forty BALB/c mice were randomly divided into normal group,model group,Yulian Pills group(0.5 g·kg-1)and Mesalazine group(0.3 g·kg-1),10 mice in each group.The mouse model of ulcerative colitis was induced by ad libitum drinking 3%DSS solution for 7 days.During the experiment,the mental state,faecal characteristics,blood in stool and body mass of the mice were recorded daily,and the length and mass of the colon were measured and the colon mass index was calculated;HE staining was used to observe the pathological and morphological changes of the colon tissue;ELISA was used to determine the expression levels of interleukin(IL)-6 and IL-15 in the colon tissues;flow cytometry was used to determine the mTfh cell subpopulation in the spleen tissue expression;Western Blot was used to determine the protein expression levels of Roquin-1,AMPK-α,p-AMPK-α in colon tissues.Results Compared with the normal group,the mice in the model group showed a significant decrease in body mass(P<0.01),a significant shortening of colon length(P<0.01),significant increase in colon mass(P<0.05)and colon mass index(P<0.01),and severe pathological damage to colon tissues;the expression levels of the pro-inflammatory cytokines IL-6 and IL-15 in the colon tissues were significantly increased(P<0.01);cell expression levels of CD4+CCR7-CXCR5+CD62L+,CD4+CCR7+CXCR5+ GL7+,CD4+CCR7-CXCR5+GL7+ were significantly increased in spleen tissues(P<0.01),whereas the expression level of CD4+CCR7+CXCR5+CD62L+ cell was significantly decreased(P<0.01);and protein expression levels of Roquin-1,AMPK-α,p-AMPK-α were significantly reduced in the colonic tissues(P<0.05).Compared with the model group,mice in the Yulian Pills group and Mesalazine group showed a significant increase in body mass(P<0.05),a significant extension of colon length(P<0.01),a significant reduction in colon mass(P<0.05),a significant decrease in the colon mass index(P<0.01),and a more obvious improvement in pathological damage of the colon tissues;a significant decrease in the expression levels of IL-6 and IL-15 in the colon tissues(P<0.01);cell expression levels of CD4+CCR7-CXCR5+CD62L+,CD4+CCR7+CXCR5+GL7+,CD4+CCR7-CXCR5+GL7+ in splenic tissues was significantly reduced(P<0.01),whereas the expression level of CD4+CCR7+CXCR5+CD62L+ cell was significantly increased(P<0.01);the protein expression levels of Roquin-1,AMPK-α,and p-AMPK-α were significantly increased in colon tissues(P<0.05,P<0.01).Conclusion The therapeutic effect of Yulian Pills on DSS-induced ulcerative colitis mice can ameliorate the histopathological damage of colon,which may be related to the activation of the Roquin-1/MPK-α signalling pathway,the down-regulation of the expressions of inflammatory cytokines IL-6 and IL-15,and the modulation of the homeostasis of the mTfh cell subpopulation.

Yulian Pillsulcerative colitismTfh cellsRoquin-1/AMPK-α signaling pathwaymice

周文、张哲言、黄莉、邓柏龄、刘端勇、陈文潇、赵海梅

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南昌医学院/江西省卫生健康药效与安全性评价重点实验室,江西 南昌 330052

江西中医药大学研究生院,江西 南昌 330004

江西中医药大学中医学院,江西 南昌 330004

江西中医药大学方证研究中心,江西 南昌 330004

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萸连丸 溃疡性结肠炎 记忆性滤泡辅助性T细胞 Roquin-1/AMPK-α信号通路 小鼠

国家自然科学基金项目江西省教育厅科技计划项目江西省教育厅科技计划项目江西省卫生健康委员会科技计划项目江西中医药大学首批校级科技创新团队——方证对应免疫代谢创新团队项目

82060799GJJ218903GJJ218919SKJP_220211032CXTD22008

2024

中药新药与临床药理
广州中医药大学

中药新药与临床药理

CSTPCD北大核心
影响因子:0.908
ISSN:1003-9783
年,卷(期):2024.35(2)
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