益气活络方通过调控JAK2/STAT3通路对类风湿关节炎模型大鼠Th17/Treg平衡的影响

doi:10.12092/j.issn.1009-2501.2026.05.001

中国临床药理学与治疗学 ›› 2026, Vol. 31 ›› Issue (5): 577-584.doi: 10.12092/j.issn.1009-2501.2026.05.001

益气活络方通过调控JAK2/STAT3通路对类风湿关节炎模型大鼠Th17/Treg平衡的影响

张浩¹(), 李浩林¹, 陈平¹, 王爱华¹, 程伟刚², 柏倩², 杨娟娟¹, 杨玉萍², 张智敏², 王海东¹^,*()

1. 甘肃省中医院，兰州 730050，甘肃
2. 甘肃中医药大学，兰州 730000，甘肃

收稿日期:2024-11-15 修回日期:2025-12-08 出版日期:2026-05-26 发布日期:2026-06-02
通讯作者: 王海东 E-mail:505695690@qq.com;whaidong5895@163.com
作者简介:张浩，男，主治医师，研究方向：风湿病的中医药防治研究。E-mail：505695690@qq.com
基金资助:
国家中医药管理局高水平重点学科建设项目（国中医药人教函[2022]226号）；国家自然科学基金项目（82260880）；甘肃省中医药科研项目（GZKG-2025-65）

Effect of Yiqi Huoluo Formula on Th17/Treg balance in rheumatoid arthritis model rats via regulating the JAK2/STAT3 pathway

Hao ZHANG¹(), Haolin LI¹, Ping CHEN¹, Aihua WANG¹, Weigang CHENG², Qian BAI², Juanjuan YANG¹, Yuping YANG², Zhimin ZHANG², Haidong WANG¹^,*()

1. Gansu Provincial Hospital of Traditional Chinese Medicine, Lanzhou 730050, Gansu, China
2. Gansu University of Chinese Medicine, Lanzhou 730000, Gansu, China

Received:2024-11-15 Revised:2025-12-08 Online:2026-05-26 Published:2026-06-02
Contact: Haidong WANG E-mail:505695690@qq.com;whaidong5895@163.com

摘要/Abstract

摘要：

目的: 探讨益气活络方对类风湿关节炎模型（CIA）大鼠的治疗作用及其调控JAK2/STAT3通路对Th17/Treg平衡的影响。方法: 采用牛Ⅱ型胶原乳剂诱导法建立CIA模型大鼠，以枸橼酸托法替布作为阳性对照，经不同剂量益气活络方干预28 d后观察大鼠关节肿胀程度和关节炎指数评分；HE染色观察关节病理形态学变化；ELISA法检测血清中IL-17、IL-6、TNF-α和IL-10水平；流式细胞术检测外周血中Th17和Treg比例；qRT-PCR检测滑膜组织RORγt、Foxp3 mRNA表达；Western blot检测滑膜组织中RORγt、Foxp3、p-JAK2、p-STAT3蛋白表达。结果: 益气活络方能够有效降低CIA大鼠关节肿胀程度和关节炎指数评分（P<0.01），改善大鼠关节组织病理损伤，降低大鼠血清中IL-17、IL-6、TNF-α含量和外周血中Th17/Treg比例并升高IL-10水平（P<0.05，P<0.01）；降低大鼠滑膜组织RORγt mRNA及蛋白表达水平，升高Foxp3 mRNA及蛋白表达水平（P<0.05，P<0.01）；此外还能够显著抑制滑膜组织中p-JAK2、p-STAT3蛋白表达水平（P<0.05，P<0.01）。结论: 益气活络方能够通过抑制JAK2/STAT3信号通路调控Th17/Treg平衡，显著减轻CIA模型大鼠的关节炎症状。

关键词: 益气活络方, 类风湿关节炎, JAK2/STAT3通路, Th17/Treg平衡

Abstract:

AIM: To investigate the therapeutic effect of Yiqi Huoluo Formula on collagen-induced arthritis (CIA) rat models and its influence on the Th17/Treg balance by regulating the JAK2/STAT3 pathway. METHODS: The CIA rat models were established using bovine type II collagen emulsion induction, with tofacitinib citrate serving as the positive control. After 28 days of intervention with different doses of Yiqi Huoluo Formula, the degree of joint swelling and the arthritis index scores of the rats were observed. Pathological and morphological changes in the joints were evaluated using HE staining. The serum levels of IL-17, IL-6, TNF-α, and IL-10 were detected by ELISA. The proportions of Th17 and Treg cells in peripheral blood were measured by flow cytometry. The mRNA expression levels of RORγt and Foxp3 in synovial tissues were detected by qRT-PCR. The protein expression levels of RORγt, Foxp3, p-JAK2, and p-STAT3 in synovial tissues were determined by Western blot. RESULTS: Yiqi Huoluo Formula effectively reduced the degree of joint swelling and the arthritis index scores in CIA rats (P<0.01), and improved the histopathological damage of the joint tissues. It decreased the serum levels of IL-17, IL-6, and TNF-α, as well as the Th17/Treg ratio in peripheral blood, while increasing the IL-10 levels (P<0.05, P<0.01). The formula down-regulated the mRNA and protein expression levels of RORγt and up-regulated the mRNA and protein expression levels of Foxp3 in the synovial tissues of rats (P<0.05, P<0.01). In addition, it significantly inhibited the protein expression levels of p-JAK2 and p-STAT3 in synovial tissues (P<0.05, P<0.01). CONCLUSION: Yiqi Huoluo Formula can significantly alleviate arthritis symptoms in CIA model rats by inhibiting the JAK2/STAT3 signaling pathway and thereby regulating the Th17/Treg balance.

Key words: Yiqi Huoluo Formula, rheumatoid arthritis, JAK2/STAT3 pathway, Th17/Treg balance

中图分类号:

张浩, 李浩林, 陈平, 王爱华, 程伟刚, 柏倩, 杨娟娟, 杨玉萍, 张智敏, 王海东. 益气活络方通过调控JAK2/STAT3通路对类风湿关节炎模型大鼠Th17/Treg平衡的影响[J]. 中国临床药理学与治疗学, 2026, 31(5): 577-584.

Hao ZHANG, Haolin LI, Ping CHEN, Aihua WANG, Weigang CHENG, Qian BAI, Juanjuan YANG, Yuping YANG, Zhimin ZHANG, Haidong WANG. Effect of Yiqi Huoluo Formula on Th17/Treg balance in rheumatoid arthritis model rats via regulating the JAK2/STAT3 pathway[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2026, 31(5): 577-584.

图/表 10

表 1

Table 1 Primer sequences

Primer name	Primer sequence 5'-3'	PCR product length (bp)
GAPDH	F:CGACCCCTTCATTGACCTC	129
GAPDH	R:GCCAGTAGACTCCACGACATAC	129
RORγt	F:GCGGCTCTCAGGCTTCAT	104
RORγt	R:TGACGAGGACGACTTCCATTG	104
Foxp3	F:AGGCAGAGGACACTCAATGAAA	155
Foxp3	R:TCTCCACTCGCACAAAGCAC	155
JAK2	F:AAGACTGCCGAGACCAGAAGG	143
JAK2	R:GGATTTGATCCACCCGAAGG	143
STAT3	F:TGGAAATAACGGGGAAGGC	172
STAT3	R:TGGAAATAACGGGGAAGGC	172

图 1

Fig.1 Arthritis index score of rats in each group ($ \overline{x} $±s, n=10) cP<0.01, compared with normal group; eP<0.05, fP<0.01, compared with model group.

图 2

Fig.2 Degree of joint swelling in each group of rats ($ \overline{x} $±s, n=10) cP<0.01, compared with normal group; eP<0.05, fP<0.01, compared with model group.

图 3

Fig.3 Effects of Yiqi Huoluo Formula on the pathological histomorphology of ankle joints of rats in various groups (HE staining, 200×) A: Normal; B: Model; C: TFTB; D: YQHL-L; E: YQHL-M; F: YQHL-H.

图 4

Fig.4 Effects of Yiqi Huoluo Formula on serum levels of IL-17, IL-6, TNF-α and IL-10 in CIA rats ($ \overline{x} $±s, n=10) cP<0.01, compared with normal group; eP<0.05, fP<0.01, compared with model group.

图 5

Fig.5 Effects of Yiqi Huoluo Formula on Th17 subpopulations in CIA rats

图 6

Fig.6 Effects of Yiqi Huoluo Formula on Treg subpopulations in CIA rats

图 7

Fig.7 Effects of Yiqi Huoluo Formula on Th17/Treg balance in CIA ($ \overline{x} $±s, n=10) cP<0.01, compared with normal group; fP<0.01, compared with model group.

图 8

Fig.8 The effects of Yiqi Huoluo Formula on RORγt and Foxp3 mRNA in rat synovial tissue ($ \overline{x} $±s, n=10) cP<0.01, compared with normal group; fP<0.01, compared with model group.

图 9 cP<0.01, compared with normal group; eP<0.05, fP<0.01, compared with model group.

Fig.9 Effects of Yiqi Huoluo Formula on protein expression of RORγt, Foxp3, p-JAK2 and p-STAT3 in rat synovial tissue ($ \overline{x} $±s, n=10) cP<0.01, compared with normal group; eP<0.05, fP<0.01, compared with model group.

参考文献 24

1	Di Matteo A, Bathon JM, Emery P. Rheumatoid arthritis[J]. Lancet, 2023, 402 (10416): 2019- 2033. doi: 10.1016/S0140-6736(23)01525-8
2	Feng W, Wan X, Fan S, et al. Mechanism underlying the action of Duanteng-Yimu Tang in regulating Treg/Th17 imbalance and anti-rheumatoid arthritis[J]. Heliyon, 2023, 9 (5): e15867. doi: 10.1016/j.heliyon.2023.e15867
3	李志, 张梦莹, 朱传苗, 等. 系统性风湿性疾病CAR-T免疫疗法临床研究进展[J]. 中国临床药理学与治疗学, 2024, 29 (8): 937- 946.
4	Han L, Yan J, Li T, et al. Wutou decoction alleviates arthritis inflammation in CIA mice by regulating Treg cell stability and Treg/Th17 balance via the JAK2/STAT3 pathway[J]. J Ethnopharmacol, 2024, 334, 118463. doi: 10.1016/j.jep.2024.118463
5	Zhao Y, Luan H, Jiang H, et al. Gegen Qinlian decoction relieved DSS-induced ulcerative colitis in mice by modulating Th17/Treg cell homeostasis via suppressing IL-6/JAK2/STAT3 signaling[J]. Phytomedicine, 2021, 84, 153519. doi: 10.1016/j.phymed.2021.153519
6	Fan KJ, Wu J, Wang QS, et al. Metformin inhibits inflammation and bone destruction in collagen-induced arthritis in rats[J]. Ann Transl Med, 2020, 8 (23): 1565. doi: 10.21037/atm-20-3042
7	Zhang Q, Peng W, Wei S, et al. Guizhi-Shaoyao-Zhimu decoction possesses anti-arthritic effects on type II collagen-induced arthritis in rats via suppression of inflammatory reactions, inhibition of invasion & migration and induction of apoptosis in synovial fibroblasts[J]. Biomed Pharmacother, 2019, 118, 109367. doi: 10.1016/j.biopha.2019.109367
8	魏伟, 吴希美, 李元建. 药理实验方法学 [M]. 北京: 人民卫生出版社, 2010.
9	张铭, 吴天宇, 刘小碣, 等. 甘草附子汤加减方通过抑制GSDMD介导的焦亡治疗CIA大鼠的作用机制研究[J]. 中国病理生理杂志, 2023, 39 (2): 325- 334. doi: 10.3969/j.issn.1000-4718.2023.02.015
10	马骏, 郭影, 苏小军, 等. 祛寒逐风合剂对寒湿痹阻证类风湿关节炎大鼠及其Th17/Treg平衡的影响[J]. 时珍国医国药, 2024, 35 (8): 1823- 1827. doi: 10.3969/j.issn.1008-0805.2024.08.08
11	刘健, 万磊, 黄传兵. 脾虚致痹探讨[J]. 中华中医药杂志, 2017, 32 (6): 2440- 2444.
12	郭海明, 王建国, 张越, 等. 郭来旺从正虚邪实论治类风湿关节炎经验[J]. 中华中医药杂志, 2023, 38 (1): 218- 221.
13	刘蔚翔, 姜泉. 类风湿关节炎“湿、热、瘀”病机理论探析[J]. 中医杂志, 2020, 61 (24): 2148- 2153. doi: 10.13288/j.11-2166/r.2020.24.008
14	Zheng Y, Wei K, Jiang P, et al. Macrophage polarization in rheumatoid arthritis: signaling pathways, metabolic reprogramming, and crosstalk with synovial fibroblasts[J]. Front Immunol, 2024, 15, 1394108. doi: 10.3389/fimmu.2024.1394108
15	Ren X, Zhang M, Zhang W, et al. Yunnan Baiyao ameliorates rheumatoid arthritis in rats by shifting the Th17/Treg cell balance and preventing osteoclast differentiation[J]. Evid Based Complement Alternat Med, 2022, 2022, 3764444. doi: 10.1155/2022/3764444
16	王铁烽, 苏洁, 王伟东, 等. 三叶青调节Th17/Treg平衡抗类风湿性关节炎的作用及机制研究[J]. 中国临床药理学与治疗学, 2023, 28 (2): 138- 146. doi: 10.12092/j.issn.1009-2501.2023.02.003
17	Min HK, Kim SH, Lee JY, et al. DJ-1 controls T cell differentiation and osteoclastogenesis in rheumatoid arthritis[J]. Sci Rep, 2022, 12 (1): 12767. doi: 10.1038/s41598-022-16285-1
18	Rezaei KF, Shahgaldi S, Azimi M, et al. Emerging therapeutic potential of regulatory T (Treg) cells for rheumatoid arthritis: new insights and challenges[J]. Int Immunopharmacol, 2022, 108, 108858. doi: 10.1016/j.intimp.2022.108858
19	何莲花, 栾慧杰, 何娟, 等. 二妙散对DBA/1小鼠胶原诱导型关节炎中Th17/Treg细胞分化的影响[J]. 中国实验方剂学杂志, 2023, 29 (2): 66- 72. doi: 10.13422/j.cnki.syfjx.20222338
20	杨天宁, 李浩林, 王海东, 等. 祖师麻乙素对CIA小鼠Th17/Treg细胞失衡的影响[J]. 中国临床药理学杂志, 2023, 39 (22): 3316- 3320. doi: 10.13699/j.cnki.1001-6821.2023.22.026
21	Lu Y, Ma Q, Yu L, et al. JAK2 inhibitor ameliorates the progression of experimental autoimmune myasthenia gravis and balances Th17/Treg cells via regulating the JAK2/STAT3-AKT/mTOR signaling pathway[J]. Int Immunopharmacol, 2023, 115, 109693. doi: 10.1016/j.intimp.2023.109693
22	Deng C, Sun S, Zhang H, et al. Sappanone A attenuates rheumatoid arthritis via inhibiting PI3K/AKT/NF-κB and JAK2/STAT3 signaling pathways in vivo and in vitro [J]. Int Immunopharmacol, 2024, 143(Pt 3): 113560.
23	Yin L, Ouyang D, Lin L, et al. Salidroside regulates imbalance of Th17/Treg and promotes ischemic tolerance by targeting STAT-3 in cerebral ischemia-reperfusion injury[J]. Arch Med Sci, 2021, 17 (2): 523- 534. doi: 10.5114/aoms.2019.85349
24	Chen W, Wu J, Shi H, et al. Hepatic stellate cell coculture enables sorafenib resistance in Huh7 cells through HGF/c-Met/Akt and Jak2/Stat3 pathways[J]. Biomed Res Int, 2014, 2014, 764981. doi: 10.1155/2014/764981

[1]	陈梦琴, 凌夕, 褚强强, 刘丹彦, 王越业, 吴甜甜, 董满玲, 魏伟, 吴育晶. IgD对类风湿关节炎患者外周血诱导的单核-巨噬细胞极化、吞噬的影响[J]. 中国临床药理学与治疗学, 2026, 31(4): 443-450.
[2]	王鹏飞, 王雨薇, 金芳梅, 陈平, 李伟青, 田雪梅, 王爱华, 王海东. 益气活络方调节PI3K/Akt/mTOR通路抑制胶原诱导型关节炎大鼠滑膜炎症的作用机制[J]. 中国临床药理学与治疗学, 2026, 31(2): 213-222.
[3]	徐豫湘, 田英, 许潜, 葛子靖, 姚璐莎. 基于NLRP3/Gasdermin D/Caspase-1/Interleukin-1β焦亡通路探讨盐酸青藤碱对佐剂性关节炎大鼠的保护作用[J]. 中国临床药理学与治疗学, 2025, 30(9): 1174-1181.
[4]	贾东晔, 潘志. 线粒体稳态在类风湿关节炎致病中的作用[J]. 中国临床药理学与治疗学, 2025, 30(4): 541-547.
[5]	王培培, 潘林坤, 杨魁, 冯丹丹. 清络饮通过调控 HIF-1α/VEGF通路抑制佐剂诱导关节炎大鼠的滑膜血管新生[J]. 中国临床药理学与治疗学, 2025, 30(3): 366-373.
[6]	杨娟娟, 李浩林, 杨天宁, 程伟刚, 王振东, 金芳梅, 年芳红, 苏小军, 王家强, 王海平, 王海东. 祖师麻膏药对胶原诱导型关节炎小鼠炎症及骨破坏的抑制作用[J]. 中国临床药理学与治疗学, 2024, 29(9): 979-987.
[7]	关蕊, 姚家树, 赵夜雨, 郑建南, 齐庆, 于静, 高明利. 益气养阴通络方对类风湿关节炎的作用分析及验证[J]. 中国临床药理学与治疗学, 2024, 29(1): 26-36.
[8]	范杰, 金永明, 江孝龙, 蒋国华. lncRNA HOTAIR调控miR-206影响类风湿关节炎滑膜细胞增殖和凋亡的分子机制研究[J]. 中国临床药理学与治疗学, 2023, 28(7): 736-742.
[9]	王铁烽, 苏洁, 王伟东, 夏荣双, 李思凡, 朱文瑞, 楼招欢. 三叶青调节Th17/Treg平衡抗类风湿性关节炎的作用及机制研究[J]. 中国临床药理学与治疗学, 2023, 28(2): 138-146.
[10]	杜娟，徐玲燕，吴伟，沈青青，江振洲，张陆勇，黄鑫. 类风湿关节炎下转运体表达及活性的变化[J]. 中国临床药理学与治疗学, 2018, 23(2): 223-229.
[11]	陈兰芳，强孚勇，徐亮. 托珠单抗联合改善病情抗风湿药治疗中重度类风湿关节炎短期临床观察[J]. 中国临床药理学与治疗学, 2018, 23(12): 1386-1391.
[12]	伍嘉琪，杨俏雯，陈秀敏，黄闰月，黄清春，赵越，吴晓东. 艾拉莫德联合甲氨蝶呤治疗类风湿关节炎有效性和安全性的系统评价和Meta分析[J]. 中国临床药理学与治疗学, 2018, 23(10): 1132-1140.
[13]	杨红，林杉，杨宏，谢晓薇，王新. 艾拉莫德对类风湿关节炎患者血管内皮生长因子及色素上皮衍生因子表达的影响[J]. 中国临床药理学与治疗学, 2017, 22(1): 68-71.
[14]	李小心, 王田玲, 余菲, 邵体红, 陈兰芳, 毛桐俊, 陆进明, 徐亮. 类风湿关节炎缓解后维持治疗方案的研究[J]. 中国临床药理学与治疗学, 2015, 20(7): 797-802.
[15]	刘东阳, 苏金梅, 宋瀚麟, 陈嘉, 刘洋, 幺雪婷, 宋玲, 江骥, 胡蓓. 甲氨蝶呤及其活性代谢产物在类风湿关节炎患者体内的药动学模型研究[J]. 中国临床药理学与治疗学, 2015, 20(5): 571-577.

益气活络方通过调控JAK2/STAT3通路对类风湿关节炎模型大鼠Th17/Treg平衡的影响

Effect of Yiqi Huoluo Formula on Th17/Treg balance in rheumatoid arthritis model rats via regulating the JAK2/STAT3 pathway

RichHTML

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

图/表 10

参考文献 24

相关文章 15

编辑推荐

Metrics

本文评价