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

doi:10.12092/j.issn.1009-2501.2026.05.001

Abstract

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

CLC Number:

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.

Figures/Tables 10

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

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.

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.

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.

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.

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

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

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.

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.

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.

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