泄浊解毒方通过调节Th17/Treg免疫平衡改善大鼠溃疡性结肠炎

doi:10.12122/j.issn.1673-4254.2026.02.11

摘要/Abstract

摘要：

目的探讨泄浊解毒方调节Th17/Treg免疫平衡改善溃疡性结肠炎（UC）的作用机制。方法将SD大鼠建立UC模型后随机分为模型组、美沙拉秦组（Mesalamine组，0.36 g/kg）、泄浊解毒方高剂量组（XZJD-H组，26.64 g/kg）、泄浊解毒方中剂量组（XZJD-M组，13.32 g/kg）和泄浊解毒方低剂量组（XZJD-L组，6.66 g/kg），10只/组，另取10只大鼠设为空白组。空白组和模型组采用生理盐水灌胃，Mesalamine组和泄浊解毒方各组按照相应的药物和剂量分别灌胃14 d。HE染色观察结肠组织病理改变；ELISA检测血清白细胞介素-6（IL-6）、IL-10、IL-17表达水平；免疫组织化学法（IHC）检测结肠组织IL-6、IL-10、IL-17的表达；免疫荧光法检测结肠组织闭合蛋白、闭锁小带蛋白1（ZO-1）的表达；流式细胞术检测结肠组织中Th17和Treg细胞的比例；RT-PCR检测维甲酸相关孤儿受体（ROR-γt）、叉状头转录因子p3（Foxp3）mRNA表达水平；Western blotting检测结肠组织RORγt、Foxp3蛋白的表达。结果与空白组比较，模型组出现结肠黏膜损伤、大量炎性细胞浸润等表现，血清IL-6、IL-17含量和结肠组织IL-6、IL-17表达升高（P<0.001），血清IL-10含量和结肠组织IL-10表达降低（P<0.001），Occludin、ZO-1表达水平降低，Th17细胞比例升高（P<0.001），Treg细胞比例降低（P<0.001），RORγt mRNA和蛋白表达增高（P<0.01），Foxp3 mRNA和蛋白降低（P<0.01）。与模型组比较，泄浊解毒方组可改善UC大鼠结肠黏膜的炎症反应，降低血清IL-6、IL-17含量，升高IL-10含量（P<0.01），降低结肠组织IL-6、IL-17蛋白的表达（P<0.01），升高IL-10蛋白的表达（P<0.01），升高Occludin、ZO-1的表达，降低结肠组织Th17细胞百分比（P<0.001），升高Treg细胞百分比（P<0.001），降低RORγt mRNA和蛋白表达（P<0.05），升高Foxp3 mRNA和蛋白表达（P<0.05）。结论泄浊解毒方可改善UC大鼠结肠黏膜的病理损伤，其作用机制可能与调节Th17/Treg免疫平衡有关。

关键词: 泄浊解毒方, 溃疡性结肠炎, 肠屏障, Th17/Treg免疫平衡

Abstract:

Objective To explore the mechanism of Xiezhuo Jiedu Recipe (XZJD) for improving ulcerative colitis (UC). Methods SD rat models of UC were randomized into 5 groups (n=10) for daily gavage with saline, mesalamine, or low-, medium- or high-dose XZJD for 14 days, with 10 normal rats with saline gavage as the control group. Pathological changes in the colon of the rats were observed using HE staining, and serum IL-6, IL-10 and IL-17 levels and their expressions in the colon tissues were determined with ELISA and immunohistochemistry. The expressions of occludin and ZO-1 and percentages of Th17 and Treg cells in the colon tissues were detected using immunofluorescence staining and flow cytometry; the mRNA and protein expressions of ROR-γt and Foxp3 were detected with RT-PCR and Western blotting. Results The rat models of UC showed obvious colonic mucosal damage with massive inflammatory cell infiltration, increased IL-6 and IL-17 levels and lowered IL-10 level in both the serum and colonic tissues, reduced expression levels of occludin and ZO-1, increased Th17 and lowered Treg cell percentages, increased RORγt mRNA and protein expressions, and reduced Foxp3 mRNA and protein expressions in the colonic tissue. Treatment with XZJD significantly alleviated inflammatory response in the colonic mucosa and effectively reversed the changes in IL-6, IL-17 and IL-10 levels, expressions of occludin and ZO-1, percentages of Th17 and Treg cells, and the expressions of RORγt and Foxp3 in the colonic tissues of UC rats. Conclusion XZJD can alleviate pathologies in the colonic mucosa in UC rats possibly by regulating Th17/Treg immune balance.

Key words: Xiezhuo Jiedu Recipe, ulcerative colitis, intestinal barrier, Th17/Treg immune balance

李斌杰, 周晓芳, 朗晓猛, 康欣, 刘建平. 泄浊解毒方通过调节Th17/Treg免疫平衡改善大鼠溃疡性结肠炎[J]. 南方医科大学学报, 2026, 46(2): 335-344.

Binjie LI, Xiaofang ZHOU, Xiaomeng LANG, Xin KANG, Jianping LIU. Xiezhuo Jiedu Recipe improves ulcerative colitis in rats by regulating Th17/Treg immune balance[J]. Journal of Southern Medical University, 2026, 46(2): 335-344.

图/表 8

表1 引物序列

Tab.1 Primer sequences for RT-qPCR

Gene	Forward (5'→3')	Reverse (3'→5')
RORγt	ACCAACCTCTTCTCACGGG	CTTCCATTGCTCCTGCTTTC
Foxp3	GCTTGTTTGCTGTGCGGAGA	TGGCATAGGTGAAAGGGGGC
GAPDH	ACAGCAACAGGGTGGTGGAC	TTTGAGGGTGCAGCGAACTT

图1 各组大鼠结肠组织病理变化

Fig.1 Pathological changes in colonic tissue of the rats in each group (HE staining).

图2 　各组大鼠IL-6、IL-17、IL-10的含量

Fig.2 　Serum levels of IL-6, IL-17 and IL-10 ineach group. *P<0.001 vs Control group; #P<0.01vs Model group,△P<0.05 vs XZJD-L group.

图3 各组大鼠结肠组织IL-6、IL-17、IL-10蛋白的表达

Fig.3 Immunohistochemical detection of expressions of IL-6, IL-17 and IL-10 proteins in the colon tissue of the rats in each group (Original magnification: ×400). *P<0.001 vs Control group; #P<0.01 vs Model group.

图4 各组大鼠结肠组织Occludin、ZO-1蛋白的比较

Fig.4 Expressions of occludin and ZO-1 proteins in the colon tissue of the rats in each group (DAPI nuclear staining, ×200).

图5 大鼠结肠组织中Th17、Treg细胞比例

Fig.5 Percentages of Th17 and Treg cells in rat colon tissues. *P<0.001 vs Control group; #P<0.001 vs Model group.

图6 大鼠结肠RORγt和Foxp3 mRNA表达的比较

Fig.6 Expression levels of RORγt and Foxp3 mRNA in rat colon tissues. *P<0.01 vs Control group;#P<0.05 vs Model group.

图7 大鼠结肠RORγt、Foxp3蛋白表达的比较

Fig.7 Expression levels of RORγt and Foxp3 proteins in rat colon tissues detected by Western blotting. *P<0.001 vs Control group; #P<0.01 vs Model group.

参考文献 56

[1]	Su L, Su Y, An Z, et al. Fermentation products of Danshen relieved dextran sulfate sodium-induced experimental ulcerative colitis in mice[J]. Sci Rep, 2021, 11(1): 16210. doi：10.1038/s41598-021-94594-7
[2]	Ungaro R, Mehandru S, Allen PB, et al. Ulcerative colitis[J]. Lancet, 2017, 389(10080): 1756-70. doi：10.1016/s0140-6736(16)32126-2
[3]	Nanki K, Fujii M, Shimokawa M, et al. Somatic inflammatory gene mutations in human ulcerative colitis epithelium[J]. Nature, 2020, 577(7789): 254-9. doi：10.1038/s41586-019-1844-5
[4]	Abelson JS, Michelassi F, Mao J, et al. Higher surgical morbidity for ulcerative colitis patients in the era of biologics[J]. Ann Surg, 2018, 268(2): 311-7. doi：10.1097/sla.0000000000002275
[5]	Jones GR, Lyons M, Plevris N, et al. IBD prevalence in Lothian, Scotland, derived by capture-recapture methodology[J]. Gut, 2019, 68(11): 1953-60. doi：10.1136/gutjnl-2019-318936
[6]	Rubin DT, Ananthakrishnan AN, Siegel CA, et al. ACG clinical guideline: ulcerative colitis in adults[J]. Am J Gastroenterol, 2019, 114(3): 384-413. doi：10.14309/ajg.0000000000000152
[7]	Chen R, Lai LA, Brentnall TA, et al. Biomarkers for colitis-associated colorectal cancer[J]. World J Gastroenterol, 2016, 22(35): 7882-91. doi：10.3748/wjg.v22.i35.7882
[8]	Zhu YF, Yang SH, Zhao N, et al. CXCL8 chemokine in ulcerative colitis[J]. Biomed Pharmacother, 2021, 138: 111427. doi：10.1016/j.biopha.2021.111427
[9]	Dubin PJ, Kolls JK. Th17 cytokines and mucosal immunity[J]. Immunol Rev, 2008, 226: 160-71. doi：10.1111/j.1600-065x.2008.00703.x
[10]	曹霞, 邱榕, 陶云平, 等. 基于Th17/Treg平衡的调节探讨参苓白术散治疗溃疡性结肠炎的疗效及作用机制[J].中药材, 2025, (1):237-41.
[11]	Duan S, Cao Y, Chen P, et al. Circulating and intestinal regulatory T cells in inflammatory bowel disease: a systemic review and meta-analysis[J]. Int Rev Immunol, 2024, 43(2): 83-94. doi：10.1080/08830185.2023.2249525
[12]	Josefowicz SZ, Lu LF, Rudensky AY. Regulatory T cells: mechanisms of differentiation and function[J]. Annu Rev Immunol, 2012, 30: 531-64. doi：10.1146/annurev.immunol.25.022106.141623
[13]	Ueno A, Jeffery L, Kobayashi T, et al. Th17 plasticity and its relevance to inflammatory bowel disease[J]. J Autoimmun, 2018, 87: 38-49. doi：10.1016/j.jaut.2017.12.004
[14]	覃小燕, 韩晓玲, 张媛, 等.基于网络药理学的女贞子抗骨质疏松作用机制研究[J].中国药学杂志, 2021, 56(5): 368-78.
[15]	任娟, 张娜, 王敏, 等. 基于网络药理学的砂仁镇痛作用机制研究[J].中国药学杂志, 2021, 56(9): 723-30.
[16]	Xiang SY, Zhao J, Lu Y, et al. Network pharmacology-based identification for therapeutic mechanism of Ling-Gui-Zhu-Gan decoction in the metabolic syndrome induced by antipsychotic drugs[J]. Comput Biol Med, 2019, 110: 1-7. doi：10.1016/j.compbiomed.2019.05.007
[17]	Zhu BC, Zhang WT, Lu Y, et al. Network pharmacology-based identification of protective mechanism of Panax Notoginseng Saponins on aspirin induced gastrointestinal injury[J]. Biomed Pharmacother, 2018, 105: 159-66. doi：10.1016/j.biopha.2018.04.054
[18]	Wei Y, Ren S, Wang R, et al. Based on network pharmacology to explore the potential bioactive compounds and mechanisms of Zuojin pill for the treatment of ulcerative colitis[J]. Evid Based Complement Alternat Med, 2021, 2021: 7567025. doi：10.1155/2021/7567025
[19]	李斌杰, 康帅, 刘晓萌, 等. 基于网络药理学和实验验证探讨泄浊解毒方治疗溃疡性结肠炎的作用机制[J]. 中国药学杂志, 2023, 58(1):48-56.
[20]	刘建平, 吴鹤伶, 郎晓猛, 等. 泄浊解毒方对溃疡性结肠炎大鼠血清IL-1β、IL-10及结肠黏膜CD14的影响[J]. 河北中医药学报, 2017,32(2): 1-4.
[21]	康欣, 刘建平, 任杰, 等. 泄浊解毒方改善大鼠溃疡性结肠炎和调控巨噬细胞极化机制研究[J].免疫学杂志, 2024, 40(1): 65-71.
[22]	Morris GP, Beck PL, Herridge MS, et al. Hapten-induced model of chronic inflammation and ulceration in the rat colon[J]. Gastroenterology, 1989, 96(3): 795-803. doi：10.1016/s0016-5085(89)80079-4
[23]	Sun M, He C, Wu W, et al. Hypoxia inducible factor-1α-induced interleukin-33 expression in intestinal epithelia contributes to mucosal homeostasis in inflammatory bowel disease[J]. Clin Exp Immunol, 2017, 187(3): 428-40. doi：10.1111/cei.12896
[24]	徐叔云. 药理实验方法学[M]. 3版. 北京: 人民卫生出版社, 2002.
[25]	Danese S, Fiocchi C. Ulcerative colitis[J]. N Engl J Med, 2011, 365(18): 1713-25. doi：10.1056/nejmra1102942
[26]	Kakuta Y, Ichikawa R, Fuyuno Y, et al. An integrated genomic and transcriptomic analysis reveals candidates of susceptibility genes for Crohn's disease in Japanese populations[J]. Sci Rep, 2020, 10(1): 10236. doi：10.1038/s41598-020-66951-5
[27]	娄莹莹, 李佃贵, 霍永利, 等. 溃疡性结肠炎特色病机“浊毒损膜伤络”及其意义[J]. 中国中西医结合杂志, 2022, 42(6): 749-53.
[28]	郎晓猛, 刘建平, 李佃贵, 等. 泄浊解毒方对溃疡性结肠炎大鼠结肠组织TLR2基因、蛋白表达的影响[J]. 中华中医药杂志, 2017, 32(2):838-40.
[29]	李雪可. 泄浊解毒方联合美沙拉秦治疗UC患者临床疗效观察及对肠道菌群的影响[D]. 河北中医学院, 2021.
[30]	康欣, 刘建平, 李佃贵, 等.泄浊解毒方联合美沙拉嗪治疗溃疡性结肠炎90例临床疗效观察[J].世界中西医结合杂志, 2019, 14(1): 90-3.
[31]	Herndon JS, Vitta S, Weber FH. Marked eosinophilia in a 27-year-old woman with recent onset ulcerative colitis[J]. Gastroenterology, 2021, 160(1): 29-30. doi：10.1053/j.gastro.2020.08.030
[32]	Fu X, Sun F, Sun F, et al. Aloperine protects mice against DSS-induced colitis by PP2A-mediated PI3K/Akt/mTOR signaling suppression[J]. Mediators Inflamm, 2017, 2017: 5706152. doi：10.1155/2017/5706152
[33]	Afif W, Sandborn WJ, Faubion WA, et al. Risk factors for lymphoma in patients with inflammatory bowel disease: a case-control study[J]. Inflamm Bowel Dis, 2013, 19(7): 1384-9. doi：10.1097/mib.0b013e318281325e
[34]	Shahini A, Shahini A. Role of interleukin-6-mediated inflammation in the pathogenesis of inflammatory bowel disease: focus on the available therapeutic approaches and gut microbiome[J]. J Cell Commun Signal, 2023, 17(1): 55-74. doi：10.1007/s12079-022-00695-x
[35]	Machiels K, Joossens M, Sabino J, et al. A decrease of the butyrate-producing species Roseburia hominisand Faecalibacterium prausnitzii defines dysbiosis in patients with ulcerative colitis[J]. Gut, 2014, 63(8): 1275-83. doi：10.1136/gutjnl-2013-304833
[36]	Pankratz S, Ruck T, Meuth SG, et al. CD4⁺HLA-G(+) regulatory T cells: Molecular signature and pathophysiological relevance[J]. Hum Immunol, 2016, 77(9): 727-33. doi：10.1016/j.humimm.2016.01.016
[37]	Jofra T, Galvani G, Cosorich I,et al.Experimental colitis in IL-10-deficient mice ameliorates in the absence of PTPN22[J]. Clin Exp Immunol, 2019, 197(3):263-75. doi：10.1111/cei.13339
[38]	于斌,邓力,张丽,等.湿邪致病现代机理研究进展[J].广州中医药大学学报,2015,32(1):174-7.
[39]	田同德,杨峰,唐静雯,等.慢性炎性反应与胃癌前病变关系及中医对策探讨[J].中华中医药杂志,2016,31(2):359-63.
[40]	Wu H, Chen QY, Wang WZ, et al. Compound sophorae decoction enhances intestinal barrier function of dextran sodium sulfate induced colitis via regulating Notch signaling pathway in mice[J]. Biomed Pharmacother, 2021, 133: 110937. doi：10.1016/j.biopha.2020.110937
[41]	Yin SJ, Yang HF, Tao Y, et al. Artesunate ameliorates DSS-induced ulcerative colitis by protecting intestinal barrier and inhibiting inflammatory response[J]. Inflammation, 2020, 43(2): 765-76. doi：10.1007/s10753-019-01164-1
[42]	Chu XQ, Wang J, Chen GX, et al. Overexpression of microRNA-495 improves the intestinal mucosal barrier function by targeting STAT3 via inhibition of the JAK/STAT3 signaling pathway in a mouse model of ulcerative colitis[J]. Pathol Res Pract, 2018, 214(1): 151-62. doi：10.1016/j.prp.2017.10.003
[43]	Yang F, Wang A, Zeng X, et al. Lactobacillus reuteri I5007 modulates tight junction protein expression in IPEC-J2 cells with LPS stimulation and in newborn piglets under normal conditions[J]. BMC Microbiol, 2015, 15: 32. doi：10.1186/s12866-015-0372-1
[44]	Tatiya-Aphiradee N, Chatuphonprasert W, Jarukamjorn K. Immune response and inflammatory pathway of ulcerative colitis[J]. J Basic Clin Physiol Pharmacol, 2018, 30(1): 1-10. doi：10.1515/jbcpp-2018-0036
[45]	Iacomino G, Rotondi Aufiero V, Iannaccone N, et al. IBD: Role of intestinal compartments in the mucosal immune response[J]. Immunobiology, 2020, 225(1): 151849. doi：10.1016/j.imbio.2019.09.008
[46]	Gomez-Bris R, Saez A, Herrero-Fernandez B, et al. CD4 T-cell subsets and the pathophysiology of inflammatory bowel disease[J]. Int J Mol Sci, 2023, 24(3): 2696. doi：10.3390/ijms24032696
[47]	Zhao Q, Duck LW, Huang F, et al. CD4⁺ T cell activation and concomitant mTOR metabolic inhibition can ablate microbiota-specific memory cells and prevent colitis[J]. Sci Immunol, 2020, 5(54): eabc6373. doi：10.1126/sciimmunol.abc6373
[48]	Xiong XY, Cheng Z, Wu F, et al. Berberine in the treatment of ulcerative colitis: a possible pathway through Tuft cells[J]. Biomed Pharmacother, 2021, 134: 111129. doi：10.1016/j.biopha.2020.111129
[49]	Gupta PK, Wagner SR, Wu Q, et al. Th17 cells are not required for maintenance of IL-17A-producing γδ T cells in vivo [J]. Immunol Cell Biol, 2017, 95(3): 280-6. doi：10.1038/icb.2016.94
[50]	Thapa B, Pak S, Kwon HJ, et al. Decursinol angelate ameliorates dextran sodium sulfate-induced colitis by modulating type 17 helper T cell responses[J]. Biomol Ther: Seoul, 2019, 27(5): 466-73. doi：10.4062/biomolther.2019.004
[51]	Ivanov II, McKenzie BS, Zhou L, et al. The orphan nuclear receptor RORgammat directs the differentiation program of proinflammatory IL-17+ T helper cells[J]. Cell, 2006, 126(6): 1121-33. doi：10.1016/j.cell.2006.07.035
[52]	Chen X, Du Y, Lin XQ, et al. CD4⁺CD25⁺ regulatory T cells in tumor immunity[J]. Int Immunopharmacol, 2016, 34: 244-9. doi：10.1016/j.intimp.2016.03.009
[53]	Yan JB, Luo MM, Chen ZY, et al. The function and role of the Th17/treg cell balance in inflammatory bowel disease[J]. J Immunol Res, 2020: 8813558. doi：10.1155/2020/8813558
[54]	Barbi J, Pardoll DM, Pan F. Ubiquitin-dependent regulation of Foxp3 and Treg function[J]. Immunol Rev, 2015, 266(1): 27-45. doi：10.1111/imr.12312
[55]	Nie J, Li YY, Zheng SG, et al. FOXP3⁺ Treg cells and gender bias in autoimmune diseases[J]. Front Immunol, 2015, 6: 493. doi：10.3389/fimmu.2015.00493
[56]	Sheng Y, Wu T, Dai Y, et al. The effect of 6-gingerol on inflammatory response and Th17/Treg balance in DSS-induced ulcerative colitis mice[J]. Ann Transl Med, 2020, 8(7): 442. doi：10.21037/atm.2020.03.141