The Mechanism Of Wenshen Xiaozheng Decoction In Treating Fibrosis Of Endometriosis From The Perspective Of M2 Macrophage Activation

Objective:1.To establish the mice model of endometriosis,to explore the relationship between M2 macrophages and fibrosis of endometriosis.2.To observe the effects of drugs on the degree of fibrosis in ectopic lesions of mice by using different doses of Wenshen Xiaozheng Decoction in mice with endometriosis.The levels of TGF-? and the expression of CD206 and CD68 in the mice were detected.To explore the mechanism of Wenshen Xiaozheng Decoction to improve the degree of fibrosis,and to provide scientific basis for the clinical use of Wenshen Xiaozheng Decoction in the treatment of endometriosis.Methods:Part ?1.BALB/c mice endometriosis model was established by intraperitoneal injection.The growth and histomorphology of ectopic lesion were observed on 7th,14th and 21th day after modeling.2.Masson staining was used to observe the degree of fibrosis of ectopic lesion.The area ratio of collagen fibers in each group was analyzed by Image-Pro-Plus 6.0(IPP6.0).3.Immunohistochemistry was used to observe the expression of CD206 and CD68 in ectopic lesion and eutopic endometrium.The results of immunohistochemistry were analyzed by IPP6.0.Part ?1.In addition to Blank group,the other mice using intraperitoneal injection method to establish endometriosis model.After 21 days,The rats were randomly divided into Model group,Mifepristone group,Sanjie Zhentong group,Wenshen Xiaozheng Decoction high,middle and low dose group,mice were given intragastric administration.Blank group and Model group given equal volume Distilled water.Each group of mice fed once a day,continue 28days.2.After 28 days,the mice were sacrificed to observe the growth and histomorphology of ectopic lesions.The expression of TGF-(3 in mice was detected by enzyme linked immunosorbent assay(ELISA),The area ratio of collagen fibers was observed by Masson staining.The expression of CD206 and CD68 was detected by immunohistochemistry,all the results of statistical analysis.Results:1.The success rate of the establishment of endometriosis model by intraperitoneai injection was 100%,mice model of endometriosis with the passage of time,the lesion volume increased gradually,the pelvic adhesion degree also increased gradually.2.There was a positive correlation between the fibrosis and the expression of M2 macrophages in endometriosis model mice.3.Effects of drugs on the volume of mice in each group:The volume of ecropic lesion in Model group was larger than that in other groups,which was statistically significant(P<0.05).The volume of ectopic lesion in Mifepristone group was significantly smaller than that in Model group(P<0.01).4.Effects of drugs on the area ratio of collagen fibers of mice in each group:The area ratio of collagen fibers in the Model group was significantly higher than that in the other groups(P<0.01),There was no significant difference between the the Middle-dose group of Wenshen Xiaozheng Decoction,Model group with Black group on area ratio of collagen fibers(P>0.05),The area ratio of collagen fibers in the Middle-dose group of Wenshen Xiaozheng Decoction was lower than that in High-dosed group and Low-dose group(P<0.05).5.Effects of drugs on the levels of TGF-? of mice in each group:The levels of TGF-P in the Model group were significantly higher than in the other groups(P<0.05).The levels of TGF-? in the Middle-dose group of Wenshen Xiaozheng Decoction were lower than those in Sanjie Zhentong decoction group,High-dosed group and Low-dose group(P<0.05).There was no significant difference in the level of TGF-? between the Middle-dose group of Wenshen Xiaozheng Decoction and the Mifepristone group(P>0.05).6.Effects of drugs on the levels of CD206 and CD68 of mice in each group:CD206 and CD68 were expressed in all the ectopic lesions of mice.The MOD values of CD206 and CD68 in Blank group were lower than in other groups,which were statistically significant(P<0.05),The MOD values of CD206 and CD68 in Model group were higher than in other groups(P<0.05).There were no significant differences in the MOD values of CD206 and CD68 in Mifepristone group,Sanjie Zhentong decoction group,High-dose,Middle-dose,Low-dose group of Wenshen Xiaozheng Decoction group(P<0.05),Conclusion:1.Intraperitoneal injection method can successfully establish a mice model of endometriosis,the success rate is high.2.There was a positive correlation between the fibrosis and expression of M2 macrophages in endometriosis model mice.3.Wenshen Xiaozheng decoction can reduce the level of TGF-?,the collagen deposition,significantly reduce the degree of fibrosis in mice with endometriosis.4.Wenshen Xiaozheng Decoction improves the fibrosis of endornetriosis by reducing M2 macrophages in mice with endometriosis.