Effect of Wudan Pill on Polarization of Macrophages in Endometriosis with Cold Congeal and Blood Stasis Syndrome Based on p38 MAPK Pathway
10.13422/j.cnki.syfjx.20222428
- VernacularTitle:基于p38 MAPK通路探讨乌丹丸对子宫内膜异位症寒凝血瘀证巨噬细胞极化的影响
- Author:
Jie ZHU
1
;
Xiaoou XUE
1
;
Zhiping HE
1
;
Yong DAI
1
;
Jun LI
1
;
Jiawei ZHANG
1
;
Haiyun SUN
1
Author Information
1. Dongzhimen Hospital， Beijing University of Chinese Medicine， Beijing 100700， China
- Publication Type:Journal Article
- Keywords:
endometriosis;
cold congeal and blood stasis;
inflammatory factors;
macrophage polarization;
p38 mitogen activated protein kinases
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2022;28(24):105-113
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo investigate the effect of Wudan pill on the polarization of macrophages in the rat model of endometriosis （EMT） with cold congeal and blood stasis syndrome based on p38 mitogen-activated protein kinases （p38 MAPK）. MethodFemale SD rats with regular motility cycles were selected and randomly divided into sham-operated group， model group， Wudan pill high， medium， and low-dose groups （2.4， 1.2， and 0.6 g⋅kg-1）， Chinese patent medicine group， and western medicine group by the random number table method. The method of ice water bath + autologous endometrial transplantation was used to establish the rat model of EMT with cold congeal and blood stasis， and the rats were executed after 4 weeks of continuous drug administration to collect materials. Expression levels of serum tumor necrosis factor-α （TNF-α）， interleukin-1β （IL-1β）， IL-4， and transforming growth factor-β （TGF-β） were determined by enzyme-linked immunosorbent assay （ELISA） to assess inflammation. The real-time quantitative polymerase chain reaction （Real-time PCR） was performed to determine the inducible nitric oxide synthase （iNOS）， TNF-α， arginase 1 （Arg1）， and human mannose receptor （CD206） transcriptional levels to evaluate macrophage polarization. Western blot （WB） and immunofluorescence （IF） assays were used to determine the protein expression levels of iNOS and Arg1 to corroborate macrophage polarization. WB and Real-time PCR were used to determine the protein expression levels of the p38 MAPK pathway. ResultAs compared with sham-operated group， the levels of serum TNF-α， IL-1β， TGF-β， and IL-4 of rats in the model group were significantly higher （P<0.05）. In the model group， the protein levels of iNOS， TNF-α， p-p38 MAPK， and phosphorylated-extracellular signal-regulated kinase （p-ERK） in endothelial tissues were significantly higher， the mRNA levels of iNOS， TNF-α， MAPK， and ERK were significantly higher， and the mRNA and protein expression levels of Arg1 and CD206 were significantly lower （P<0.05， P<0.01）. The number of iNOS positive cells in endothelial tissues was significantly increased， and the number of Arg1 positive cells in endothelial tissues in the model group was significantly decreased （P<0.05， P<0.01）. As compared with the model group， the expression of TNF-α， IL-1β， TGF-β， and IL-4 in each administration group was reduced to different degrees， which was especially significant in the Wudan pill high and medium-dose groups and the western medicine group （P<0.05）. The protein expression levels of iNOS， TNF-α， p-p38 MAPK， and p-ERK in endometrial tissues of rats in the Wudan pill high and medium-dose groups， the Chinese patent medicine group， and the western medicine group were significantly lower， the mRNA expression levels of iNOS， TNF-α， MAPK， and ERK were significantly lower， and the protein expression levels of Arg1 and CD206 were significantly higher （P<0.05， P<0.01）. The number of iNOS positive cells in endometrial tissues of rats was significantly decreased in the Wudan pill high and medium-dose groups， the Chinese patent medicine group， and the western medicine group， whereas the number of Arg1 positive cells was increased （P<0.05， P<0.01）. The low， medium， and high doses of Wudan pill were dose-dependent， and the efficacy of the Wudan pill high-dose group was similar to that of the western medicine group. ConclusionWudan pill reduces the inflammatory response in rat model of EMT with cold congeal and blood stasis syndrome and decreases expression levels of TNF-α， IL-1β， IL-4， and TGF-β， thereby prompting polarization of macrophages from M1 to M2 type. The mechanism is presumedly related to p38 MAPK signaling pathway.
