Effects of Huoxue Xiaoyi Formula （活血消异方） on Tfh Cells and the JAK/STAT Pathway in Ectopic Tissues of Ovarian Endometriosis Model Rats
10.13288/j.11-2166/r.2025.14.011
- VernacularTitle:活血消异方对卵巢型子宫内膜异位症模型大鼠异位组织中Tfh细胞及JAK/STAT通路的影响
- Author:
Weisen FAN
1
;
Yongjia ZHANG
1
;
Yaqian WANG
2
;
Hong LEI
2
;
Huiting YAN
2
;
Ruijie HOU
1
;
Xin WANG
1
;
Yu TAO
1
;
Ruihua ZHAO
1
Author Information
1. Guang'anmen Hospital，China Academy of Chinese Medical Sciences，Beijing，100053
2. The First Clinical Medical College of Shandong University of Traditional Chinese Medicine
- Publication Type:Journal Article
- Keywords:
ovarian endometriosis;
Huoxue Xiaoyi Formula （活血消异方）;
Tfh cells;
Janus kinase;
signal transducer and activator of transcription
- From:
Journal of Traditional Chinese Medicine
2025;66(14):1473-1480
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo explore the potential mechanism of Huoxue Xiaoyi Formula （活血消异方， HXF） in treating ovarian endometriosis （OEM） from the perspective of T follicular helper （Tfh） cells and the Janus kinase/signal transducer and activator of transcription （JAK/STAT） signaling pathway. MethodsForty-five female SD rats with normal estrous cycles were randomly divided into three groups， HXF group， model group， and normal group， with 15 rats in each group. A rat model of OEM was established by autologous endometrial tissue implantation. After successful modeling， the treatment group received HXF at 5.85 g/（kg·d） by gavage for 14 consecutive days. The model group and normal group received 1 mL/d of normal saline by gavage. RNA-sequencing data from human proliferative-phase endometriotic and normal endometrial tissues were downloaded from the GEO database. Transcriptomic sequencing was used to analyze gene expression in rat ovarian ectopic tissues and normal uterine tissues， and comparisons were made with human data to verify JAK/STAT pathway activation in proliferative-phase ectopic tissues. Immunohistochemistry was used to detect the positive expression of CXC chemokine receptor 5 （CXCR5） and interleukin-21 （IL-21） in rat ovarian ectopic and normal uterine tissues. Western Blotting was performed to detect the protein levels of IL-21， IL-21 receptor （IL-21R）， Janus kinase 1 （JAK1）， signal transducer and activator of transcription 6 （STAT6）， and B-cell lymphoma 2 （Bcl-2）. Tfh cell infiltration was analyzed using immune cell infiltration methods. ResultsGene set enrichment analysis showed that the JAK/STAT pathway was significantly activated in human proliferative-phase endometriotic tissues compared to normal endometrial tissues. Similarly， the JAK/STAT pathway was markedly activated in rat ovarian ectopic tissues in the model group compared to the normal group， but suppressed in the HXF group compared to the model group. Compared with normal uterine tissues， ovarian ectopic tissues in the model group showed increased Tfh cell infiltration scores， higher CXCR5 and IL-21 expression， and elevated levels of IL-21， IL-21R， JAK1， STAT6， and Bcl-2 proteins. Compared with the model group， HXF group showed reduced CXCR5 and IL-21 expression and decreased protein levels of IL-21， IL-21R， JAK1， STAT6， and Bcl-2. ConclusionHXF may suppress activation of the JAK/STAT signaling pathway in ovarian endometriotic tissues by inhibiting IL-21 secretion from Tfh cells.
