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白头翁汤对DSS诱导的溃疡性结肠炎小鼠结肠组织mTORC1-STAT3-COX-2信号通路的影响

何琼姿 韦鹏 刘欢欢 张俊芝 刘彤彤 石琨群 徐艺 刘史佳

何琼姿, 韦鹏, 刘欢欢, 张俊芝, 刘彤彤, 石琨群, 徐艺, 刘史佳. 白头翁汤对DSS诱导的溃疡性结肠炎小鼠结肠组织mTORC1-STAT3-COX-2信号通路的影响[J]. 南京中医药大学学报, 2023, 39(1): 50-56. doi: 10.14148/j.issn.1672-0482.2023.0050
引用本文: 何琼姿, 韦鹏, 刘欢欢, 张俊芝, 刘彤彤, 石琨群, 徐艺, 刘史佳. 白头翁汤对DSS诱导的溃疡性结肠炎小鼠结肠组织mTORC1-STAT3-COX-2信号通路的影响[J]. 南京中医药大学学报, 2023, 39(1): 50-56. doi: 10.14148/j.issn.1672-0482.2023.0050
HE Qiong-zi, WEI Peng, LIU Huan-huan, ZHANG Jun-zhi, LIU Tong-tong, SHI Kun-qun, XU Yi, LIU Shi-jia. Effect of Baitouweng Decoction on mTORC1-STAT3-COX-2 Signaling Pathway on Colon Tissue of DSS-Induced Ulcerative Colitis Mice[J]. Journal of Nanjing University of traditional Chinese Medicine, 2023, 39(1): 50-56. doi: 10.14148/j.issn.1672-0482.2023.0050
Citation: HE Qiong-zi, WEI Peng, LIU Huan-huan, ZHANG Jun-zhi, LIU Tong-tong, SHI Kun-qun, XU Yi, LIU Shi-jia. Effect of Baitouweng Decoction on mTORC1-STAT3-COX-2 Signaling Pathway on Colon Tissue of DSS-Induced Ulcerative Colitis Mice[J]. Journal of Nanjing University of traditional Chinese Medicine, 2023, 39(1): 50-56. doi: 10.14148/j.issn.1672-0482.2023.0050

白头翁汤对DSS诱导的溃疡性结肠炎小鼠结肠组织mTORC1-STAT3-COX-2信号通路的影响

doi: 10.14148/j.issn.1672-0482.2023.0050
基金项目: 

国家自然科学基金面上项目 82074241

江苏省中医院高峰人才项目 y2021rc36

江苏省研究生科研创新实践活动项目 SJCX21_0693

详细信息
    作者简介:

    何琼姿, 女, 硕士研究生, E-mail: 1710886491@qq.com

    通讯作者:

    徐艺, 女, 主任中药师, 主要从事中医脾胃病研究, E-mail: fsyy00638@njucm.edu.cn

    刘史佳, 男, 研究员, 主要从事中药临床药理研究, E-mail: liushijia2011@163.com

  • 中图分类号: R285.5

Effect of Baitouweng Decoction on mTORC1-STAT3-COX-2 Signaling Pathway on Colon Tissue of DSS-Induced Ulcerative Colitis Mice

  • 摘要:   目的  探讨白头翁汤治疗溃疡性结肠炎(UC)的可能作用机制。  方法  40只C57BL/6雌鼠随机分为空白组、模型组、白头翁汤给药组及雷帕霉素给药组。采用3.5%葡聚糖硫酸钠(DSS)溶液构建小鼠UC模型。造模同时进行给药处理。白头翁汤组每日给予6.83 g·kg-1白头翁汤灌胃, 雷帕霉素组每日用2 mg·kg-1雷帕霉素药液进行腹腔注射, 空白组和模型组每日给予等体积0.5%羧甲基纤维素钠溶液灌胃。记录小鼠每日体质量变化及粪便性状, 7 d后处死小鼠。测量结肠长度并观察结肠组织病理学变化。采用流式细胞术检测小鼠血清炎症因子含量, 采用免疫印迹法检测结肠组织中哺乳动物雷帕霉素靶蛋白(mTOR)、转录激活因子3(STAT3)、核糖体S6蛋白激酶(P70S6K)及其磷酸化蛋白表达, 免疫组化法检测结肠组织中磷酸化P70S6K的表达, qPCR检测结肠组织环氧化酶(COX-2) mRNA表达水平。  结果  与空白组相比, 模型组小鼠体质量减轻, 结肠长度缩短、疾病活动指数(DAI)和组织病理学评分升高(P < 0.001), 血清中白细胞介素-6(IL-6)和肿瘤坏死因子-α(TNF-α)含量升高(P < 0.001), 结肠组织中COX-2 mRNA表达水平、p-mTOR/mTOR、p-P70S6K/P70S6K、p-STAT3/STAT3均升高(P < 0.01,P < 0.001)。与模型组相比, 白头翁汤组和雷帕霉素组小鼠上述表型症状均得到改善, 血清中IL-6和TNF-α含量下降(P < 0.01,P < 0.001), 结肠组织中p-mTOR/mTOR、p-P70S6K/P70S6K、p-STAT3/STAT3和COX-2 mRNA表达减少(P < 0.05,P < 0.01,P < 0.001)。  结论  白头翁汤能有效预防DSS诱导的小鼠结肠炎症, 其作用机制可能与抑制mTORC1-STAT3-COX-2信号通路有关。

     

  • 图  1  白头翁汤改善了DSS诱导的结肠炎症状

    注: Control.空白组; DSS.模型组; BTW.白头翁汤组; RAPA.雷帕霉素组; 与空白组比较, ###P < 0.001;与模型组比较, ***P < 0.001, **P < 0.01。

    Figure  1.  Baitouweng decoction ameliorated the symptoms of DSS-induced colitis

    图  2  各组小鼠结肠组织的代表性HE染色图(×200)

    Figure  2.  Representative hematoxylin-eosin (HE) staining of colon tissues of mice in each group (×200)

    图  3  小鼠结肠组织免疫印迹电泳图

    Figure  3.  Western blot images of mouse colon tissues

    图  4  各组小鼠结肠组织p-P70S6K免疫组化图片(×200)

    Figure  4.  The immunohistochemical staining of p-P70S6K in mouse colon tissues (×200)

    表  1  各组小鼠结肠组织病理学评分(x±s)

    Table  1.   Histological scores of colon tissues of mice in each group(x±s)

    组别 n 病理学评分
    空白组 8 0.25±0.46
    模型组 8 6.75±1.49###
    白头翁汤组 8 4.13±0.99***
    雷帕霉素组 8 4.63±1.30**
    注: 与空白组相比, ###P < 0.001;与模型组相比, ***P < 0.001, **P < 0.01。
    下载: 导出CSV

    表  2  各组小鼠血清IL-6和TNF-α的含量(x±s,pg·mL-1)

    Table  2.   Contents of IL-6 and TNF-α in mouse serum (x±s, pg·mL-1)

    组别 n IL-6 TNF-α
    空白组 5 25.72±5.90 3.61±1.26
    模型组 5 111.30±26.52### 9.12±1.94###
    白头翁汤组 5 25.56±6.44*** 3.71±0.86**
    雷帕霉素组 5 13.79±3.70*** 3.68±1.27***
    注: 与空白组相比, ###P < 0.001;与模型组相比, **P < 0.01, ***P < 0.001。
    下载: 导出CSV

    表  3  各组小鼠结肠组织p-P70S6K/P70S6K、p-STAT/STAT3和p-mTOR/mTOR值比较(x±s)

    Table  3.   Comparison of p-P70S6K/P70S6K、p-STAT/STAT3 and p-mTOR/mTOR in colon tissue of mice in each group(x±s)

    组别 n p-P70S6K/ P70S6K p-STAT3/ STAT3 p-mTOR/ mTOR
    空白组 3 0.22±0.31 0.15±0.05 0.47±0.28
    模型组 3 1.22±0.05### 1.18±0.18### 1.14±0.03##
    白头翁汤组 3 0.51±0.11** 0.58±0.21** 0.68±0.15*
    雷帕霉素组 3 0.76±0.05** 0.31±0.08*** 0.48±0.18**
    注: 与空白组相比, ##P < 0.01,###P < 0.001;与模型组相比, *P < 0.05, **P < 0.01,***P < 0.001。
    下载: 导出CSV

    表  4  各组小鼠结肠组织p-P70S6K免疫组化评分(x±s)

    Table  4.   The immunohistochemical score of p-P70S6K in mouse colon tissues(x±s)

    组别 n 组化评分
    空白组 5 1.6±0.74
    模型组 5 4.5±0.71###
    白头翁汤组 5 1.8±0.67***
    雷帕霉素组 5 1.7±0.45***
    注: 与空白组相比, ###P < 0.001;与模型组相比, ***P < 0.001。
    下载: 导出CSV

    表  5  各组小鼠结肠组织COX-2 mRNA表达情况(x±s)

    Table  5.   The mRNA level of COX-2 in mouse colon tissues (x±s)

    组别 n COX-2
    空白组 4 1.00±0.13
    模型组 4 3.96±0.79###
    白头翁汤组 4 0.47±0.18***
    雷帕霉素组 4 0.93±0.26***
    注: 与空白组相比, ###P < 0.001;与模型组相比, ***P < 0.001。
    下载: 导出CSV
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  • 收稿日期:  2022-06-08
  • 网络出版日期:  2023-01-18
  • 发布日期:  2023-01-10

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