Effects And Mechanisms Of Ethanol Extract Of Euonymus Alatus On Sequelae Of Pelvic Inflammatory Disease In Rats

Objective: Based on the pathogenesis of blood stasis and collateral obstruction in the sequelae of pelvic inflammatory diseases,the treatment of promoting blood circulation and removing blood stasis was put forward.According to the efficacy of promoting blood circulation and regulating menstruation,this research is to explore the effects and mechanisms of ethanol extract of Euonymus alatus on sequelae of pelvic inflammatory disease in rats.Methods: All the rats were adaptively fed for five days before the experiment.70 female SD rats were randomly divided into sham-operated group,model group,low,medium and high dosage group of ethanol extract of Euonymus alatus,control group A(Guizhi Fuling Capsule)and control group B(Jingangteng Capsule),10 rats in each group.Except the sham-operated group was treated with NS,the SPID model was duplicated by phenol mortar burned rat's endometrium.After the model was successfully established,the rats in the sham-operated group were given by distilled water,the other groups were fed by administration of corresponding drugs and doses.After 14 days of gavage administration,we collected the blood from rat's abdominal aorta,in addition,uterus and adnexal tissues were taken for observation and detection.After observing the uterus and fallopian tube with naked eyes,the uterus and its appendages were taken for preservation.The morphological changes of uterus were observed after HE staining;The contents of TNF-? and IL-1? in serum of rats were detected by enzyme-linked immunosorbent assay;the average optical density of ICAM-1 and VEGF in uterus was detected by immunohistochemistry;and the protein's expression of p38 MAPK was tested by Western blot.Results: 1.Morphological observation: Congestion and edema,the uterus and fallopian tube in model group rats were observed by naked eye.Some of the uterus and pelvic periphery tissues adhered,even pelvic effusion can be seen.No hyperemia,edema and adhesion were found in sham-operation group in uterus and fallopian tube tissues;Congestion,edema and adhesions of the uterus and fallopian tube in other treatment groups also improved significantly.Microscopic observation after HE staining.In model group,the uterine wall of SD rats was disordered,endometrium was congested and edema,and large number of epithelial cells proliferated,degenerated and necrotized.A great quantity of inflammatory cells infiltrated the lamina propria of uterus.Some glandular cell necrosis can be seen.Arrangement of smooth muscle cells in uterus is disordered.Suggesting that the model of SPID was very successful.The structure of uterine wall is basically clear.Epithelial cells arranged neatly,only a small number of epithelial cells degenerated and necrotized.The glandular cells are basically intact.A small amount of inflammatory cell infiltration was observed in high-dose group ethanol extract of Euonymus alatus.The structure of smooth muscle cells and serosa layer was basically normal.Endometrial inflammation in other treatment groups was significantly less than that in model group.2.Effects of TNF-? and IL-1? on serum of rats with SPID.The contents of TNF-a and IL-1? in model group were significantly higher than those in other groups(P<0.05);The contents of TNF-a and IL-1? in high-dose group,control group A(Guizhi Fuling Capsule)and sham operation group were similar(P<0.05);And it showed us a tendency of dose-dependent.3.Effects of protein expression of VEGF and ICAM-1 on uterine tissue in rats with SPID: The protein expression of ICAM-1 and VEGF in uterus of rats in model group were significantly higher than those in other treatment groups(P<0.05).The protein expression of ICAM-1 and VEGF in the high-dose group were lower than those in the model group.(P<0.05),which was close to that in control group A(Guizhi Fuling Capsule)(P>0.05).4.Effects of protein expression p38 MAPK signaling pathway-related: The expression of p-p38 MAPK in uterus in the model group was significantly higher than that of other treatment group(P<0.05);However the expression of p-p38 MAPK in high-dose group was significantly lower than that of model group(P<0.01),which was similar to that of the control group A(Guizhi Fuling Capsule)(P>0.05).Conclusion: Ethanol extract of Euonymus alatus has an obvious therapeutic effect on SPID in rats,and its level is close to that of Guizhi Fuling Capsule.It can significantly improve the inflammation of endometrial hyperemia,edema and pelvic adhesion in SPID rats,and reduce the necrosis,exfoliation and inflammatory cell infiltration of epithelial cells.Its mechanism is related to the following factors:(1)Reducing the level of pro-inflammatory cytokines in serum,inhibiting the synthesis,release and activity of pro-inflammatory cytokines is closely related,thus alleviating inflammation such as tissue congestion and edema,and promoting the repair of inflammatory tissue.(2)Reducing the gene expression level of ICAM-1,VEGF and other adhesion-related cell molecules in uterine tissue,restraining the binding between inflammatory cells and fibroblasts,thus inhibiting the formation of adhesion and scar.(3)Inhibitingof inflammatory cytokines such as TNF-?,decreasing the Activity of Adhesion-related Molecules.Restraining the activation and transmission of p38 MAPK signaling pathway,thus inhibiting the enzymatic cascade amplification of inflammatory response.