Effect of Gexia Zhuyutang on NF-κB Signaling Pathway and Downstream Factors in EMT Rats with Qi Stagnation and Blood Stasis Syndrome
10.13422/j.cnki.syfjx.20222037
- VernacularTitle:膈下逐瘀汤对EMT气滞血瘀型大鼠NF-κB信号通路及其下游因子的影响
- Author:
Zhiping WANG
1
;
Yifan CUI
1
;
Tao GAN
1
Author Information
1. Shanxi University of Chinese Medicine， Jinzhong 030619， China
- Publication Type:Journal Article
- Keywords:
Gexia Zhuyutang;
endometriosis;
nuclear factor kappa B （NF-κB） signaling pathway;
inflammatory response;
immunosuppression;
microvascular regeneration
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2022;28(22):23-32
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo investigate the therapeutic effect of Gexia Zhuyutang on rats with endometriosis （EMT） with Qi stagnation and blood stasis syndrome and its mechanism. MethodSix female unpregnant healthy SD rats of SPF grade were randomly selected as a blank group. The rest were treated with the compound factor method to induce EMT with Qi stagnation and blood stasis syndrome in rats. After successful modeling， 30 rats from 36 rats were randomly divided into a model group， a medroxyprogesterone （0.416 mg·kg-1） group， and low， medium， and high-dose Gexia Zhuyutang groups （4.063， 8.125， 16.25 g·kg-1）， with 6 rats in each group. Each group was given corresponding drugs， once a day for 4 consecutive weeks. The effect of Gexia Zhuyutang on rats with EMT was observed by the pathological changes and the volume of ectopic foci. The endometrial tissues of each group were taken for hematoxylin-eosin （HE） staining to observe histopathological changes. Enzyme-linked immunosorbent assay （ELISA） was used to determine the content of interleukin （IL）-10 and IL-6 in the serum supernatant of each group. Immunohistochemistry （IHC） was used to determine the protein expressions of nuclear factor kappa B （NF-κB）， matrix metalloproteinase-9 （MMP-9）， vascular endothelial growth factor （VEGF）， and Bcl-2 related X protein （Bax） in the tissues of rats in each group. Real-time fluorescence quantitative reverse transcription polymerase chain reaction （Real-time PCR） was used to determine the mRNA expressions of NF-κB， MMP-9， VEGF， and Bax. ResultAfter treatment， the volume of ectopic foci in the low， medium， and high-dose Gexia Zhuyutang groups and the medroxyprogesterone group was significantly reduced. The pathological observation under the microscope showed that the endometrial structure of rats in the sham-operated group and the blank group was intact， glands and mesenchymal cells grew well， glandular epithelial cells were arranged tightly and neatly， the cytoplasm was abundant and highly columnar， the interstitial cells were evenly distributed， and the blood vessels were abundant and spindle-shaped. Compared with the blank group and the sham-operated group， the model group was dominated by columnar epithelium and cubic epithelium. In the model group， ectopic endometrial epithelial cells were short columnar， some were pseudo-lamellar， the cell morphology was incomplete， the number of interstitial cells and glands was reduced， some glands were round， and degenerative dysplasia was formed. Compared with the blank group， the serum content of IL-10 in the model group was decreased （P<0.05）， and the content of IL-6 was increased （P<0.05）. The model group increased the protein expressions of NF-κB， VEGF， and MMP-9 （P<0.05） and decreased the protein expression of Bax in eutopic and ectopic endometrial tissues. Compared with the model group， the serum content of IL-10 in the medium and high-dose Gexia Zhuyutang groups was increased （P<0.05）， and the content of IL-6 was decreased （P<0.05）. The protein expressions of NF-κB， VEGF， and MMP-9 in eutopic and ectopic endometrial tissues of rats in the low， medium， and high-dose Gexia Zhuyutang were decreased （P<0.05）， and the protein expression of Bax was increased （P<0.05）. Compared with the ectopic endometrium of the model group， the mRNA expressions of NF-κB， MMP-9， and VEGF in the ectopic endometrium of the medium and high-dose Gexia Zhuyutang groups were decreased （P<0.05）， and the mRNA expression of Bax was increased （P<0.05）. ConclusionGexia Zhuyutang has the effect of inhibiting the invasion of ectopic foci in rats with EMT with Qi stagnation and blood stasis syndrome， and its mechanism may be related to the intervention of immunosuppression mediated by overactivation of the NF-κB signaling pathway， the improvement of the inflammatory response， and the blocking of microvascular regeneration function.
