| Objective: Intervention of Jilian Penqiang capsule in model rats with sequelae of pelvic inflammatory disease(SPID),was performed to observe the therapeutic effect of Jilian Penqiang capsule on SPID rats,and explore its possible mechanism focusing on fibrosis from the morphology,pathology and molecular biology,further provide theoretical basis for its clinical popularization and application.Methods: 90 female SD rats were randomly divided into normal control group(NC group,n=12)and Sham-operated group(Sham group,n=12),the remaining rats were made models by phenol mucilage method.Extracted 3 models after ten days for model validation.After the models were verified successfully by morphology and pathology observation,they were randomly divided into Model group,Fuke Qianjin capsule group(FKQJ group,n=12),Jilian Penqiang capsule low-dose,medium-dose and high-dose group(JLPQ-L,JLPQ-M,and JLPQ-H group,n=12).The NC group,Sham group and Model group were gavaged with distilled water(10 mL/kg),while the FKQJ group was gavaged with Fuke Qianjin capsule liquid(0.25 g/kg),JLPQ-L,JLPQ-M and JLPQ-H group were gavaged with Jilian Penqiang capsule liquid(0.375 g/kg,0.75 g/kg and 1.5 g/kg)per day,for 21 consecutive days.The general situation of rats was observed every day and their body weights were measured every 5~7 days.Afetr the final gavage,pelvic morphology was observed,spleen and uterus coefficients were calculated,fibrosis of uterine tissue was observed by Masson staining,and the fiber area ratio was calculated,the levels of soluble intercellular adhesion factor-1(sICAM-1)and fibroblast growth factor-2(FGF-2)in serum were detected by ELISA,the expression levels of transforming growth factor-?1(TGF-?1),transforming growth factor-?1 receptor II(T?RII),phosphorylated-Smad2/3(p-Smad2/3)and Smad7 proteins were detected by immunohistochemistry and western blot.Results: 1.Establishment of the model: After ten days of modeling,the observation results accorded with pathological characteristics of SPID,indicating that the model was established successfully.2.General situation: Before modeling,the rats in each group were in good condition.After modeling,the rats in the Model group were hyperactive and irritable at first.With the extension of modeling time,they gradually became debilitated,slowed down,reduced eating,increased drinking,and with more vaginal discharge.After intervention,the above conditions in the treatment groups were improved to varying degrees,and the JLPQ-H group was the most obvious.3.Body weight change: The weight of rats in the Model group and treatment groups decreased to the lowest on the 5th day of modeling,which were significantly different with the NC group(P<0.05).As the intervention time increased,the Model group rose least,there were differences between the Model group and the other groups(P<0.05)at the 21 st day after gavage,and the differences among the JLPQ dose groups were significant(P<0.05).4.General morphology: In the Model group,pelvic was extensive adhesion and hyperemia,severe uterine deformation,uterine cavity expansion or stenosis.All the treatment groups improved compared with the Model group.5.Spleen and uterus coefficients: The spleen and uterus coefficients of the Model group increased significantly(P<0.05),while the treatment groups notably lower than that of the Model group(P<0.05).The difference between the JLPQ-L and JLPQ-H group existed(P<0.05).6.Uterus fibrosis:(1)Qualitative analysis: In the Model group,there were few glands in the endometrium,and a large number of blue collagen fibers in the mesenchyme and muscular layer were clustered.Gland regeneration and the reduction of collagen fibers to varying degrees were observed in all treatment groups.(2)Semi-quantitative analysis: The fiber area ratio in the Model group increased notably(P<0.05).All treatment groups declined obviously when compared to the Model group(P<0.05).The differences among the various JLPQ dose groups were statistically significant(P<0.05).7.Content of sICAM-1 and FGF-2 in serum: The content of sICAM-1 and FGF-2 in the Model group increased(P<0.05),which was less in all treatment groups than that of the Model group(P<0.05),and the differences existed among the JLPQ dose groups(P<0.05).8.Expression of related proteins in the TGF-?1/Smads pathway.(1)Rough positioning and qualitative analysis: The signal and range of TGF-?1,T?RII and p-Smad2/3 particles in the Model group were stronger and wider than that in the NC group,distributed in all layers of the uterus,while the Smad7 expression was weaker.The color of TGF-?1,T?RII and p-Smad2/3 proteins in every treatment groups were lighter than that in the Model group,while the color of Smad7 was deeper,and the JLPQ-H group is closest to the NC group.(2)Semi-quantitative analysis: The expressions trend of proteins in IHC and WB were about the same.Compared with the NC group,the expression of Smad7 protein in the Model group decreased,and the expression of other proteins increased(P<0.05).After intervention,the expression of Smad7 protein in each treatment group was higher than that in the Model group,while the expression of other proteins decreased(P<0.05),and this change trend was dose-dependent in each dose group of JLPQ,the larger the dose,the more obvious the change.Conclusion: 1.Jilian Penqiang capsule can repair the tissue morphology of the model rats,alleviate swelling of spleen and uterine,improve pelvic adhesion,reduce tissue damage.2.Jilian Penqiang capsule against adhesion and fibrosis probably through regulating the levels of sICAM-1,FGF-2 in serum,as well as inhibiting pathological activation of TGF-?1/Smads signaling pathway(down-regulating the expressions of TGF-?1,T?RII and p-Smad2/3,up-regulating the expression of Smad7 in uterine tissue).3.Jilian Penqiang capsule has a dose-dependent treatment effect on SPID,the high-dose group is optimal.But most indicators have not recovered to normal levels,indicating that Jilian Penqiang capsule can only reduce the degree of tissue fibrosis damage and prevent further development of fibrosis,but the fibrosis that had been formed could not be completely reversed and eliminated.4.The efficacy and the mechanism in anti-fibrosis of Jilian Penqiang capsule and Fuke Qianjin capsule on SPID may be similar. |
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