Study On The Mechanism Of Regulating CASPASE Apoptosis Factor And MAPK Pathway In The Treatment Of EMs Rats By Neiyizhitong Decoction

Objective:By investigating the influence of Neiyi Zhitong Decoction on the histopathology and the expression of caspase-9,caspase-3,p-ERK,p-JNK and p-p38 of endometrium of EMs rats’ model,we discuss the mechanism of action of treating EMs by Neiyi Zhitong Decoction on the view of preventing the invasion and metastasis and promoting the apoptosis of cells.So that we can provide some theoretical and experimental basis for the inherit of academic experience and the scientific standardization of old and famous experts of traditional Chinese medicine(TCM).Method:We made the endometriosis building operation in SD rats by autologous grafting method.After 4 weeks,we executed the laparotomy to observe the film-forming situation.The forty rats which were successful modeling were divided into 4 groups randomly with ten rats in each group.Then these 4 groups were divided into the model group,Neiyi Zhitong Decoction low dose group,Neiyi Zhitong Decoction high dose group and western danazol group.In addition,8 rats which did not make molding were sorted into the sham operation group.All these groups were fed medicine by intragastric administration daily for 4 weeks.The dose of Neiyi Zhitong Decoction was 245.8 mg/Kg and 985.3 mg/Kg for low and high dose group respectively.The dose of danazol was 7.2 mg/Kg for the danazol group.The model group and sham group was fed 2 ml normal saline daily.We recorded the nidus size by laparotomy after using the medicine and observed the change of intimas’ histopathology of each groups’ rats by using the HE dyeing optical microscope.The caspase-9,caspase-3,p-ERK,p-JNK and p-p38 mRNA expressions in all groups were analyzed by RT-PCR.The expressions of caspase-9,caspase-3,p-ERK,p-JNK and p-p38 protein were detected by Western Blot.Results:1,The comparison of nidus size:the difference of endometrium nidus size between the medication group and model group after using the medicine was statistically significant(P<0.05).Compared with low dose Neiyi Zhitong Decoction group,the size of high dose Neiyi Zhitong Decoction group and western medicine group was reduced and more evident and the difference was statistically significant(P<0.05).Compared the high dose Neiyi Zhitong Decoction group and western medicine group,the difference was not statistically significant(P>0.05).2,On the view of morphopathological changes,in the sham operation group,the glandular epithelial cells of the eutopic endometrium were columnar,with abundant glands and complete structure;in the model group,there were vacuoles in the epithelium of ectopic foci,and the nucleus was enlarged,the arrangement was disordered,the stroma was loose,the structure of the gland was incomplete,and there were visible neovasculariztions;lastly,the pathological morphology of the drug group was close to that of the sham operation group.3,On the view of the expression of caspase-9,caspase-3 protein compared with sham operation group,the expression of protein decreased(P<0.05);compared with the model group,the expression of protein in the low group and high group and Western medicine group increased(P<0.05);In the expression of key factors of MAPK pathway,p-ERK,p-JNK and p-p38 protein,compared with the model group,the expression of p-ERK,p-JNK and p-p38 protein in high group and western danazol group decreased(P<0.05);compared with the low group,the expression of p-ERK,p-JNK and p-p38 protein in high group and western danazol group decreased(P<0.05).4,In caspase-9 and caspase-3 mRNA expression,compared with the sham operation group,the expression in the model group was decreased(P<0.05);compared with the model group,the expression in the low,high group and Western medicine group increased(P<0.05);compared with the low group,the expression in high group and western danazol group increased.In ERK,JNK,and p38 mRNA expression,compared with the model group,the expression in the low group,high group and Western medicine group decreased(P<0.05);compared with the low group,the expression in high group and western danazol group decreased(P<0.05).Conclusion:1,Neiyi Zhitong decoction can reduce endometriotic implants and improve the morphological changes.2,The unbalance of apoptosis factor caspase-9,caspase-3 and the over activation of MAPK signaling pathway play an important role in the pathogenesis of EMs.3,In the protein and mRNA level,Neiyi Zhitong decoction can increase caspase-9 and caspase-3,so it reduce the ectopic focus by promoting ectopic cell apoptosis and inhibiting the proliferation of ectopic tissue.4,Neiyi Zhitong decoction can inhibit the pathological activation of MAPK signaling pathway by reducing the expression of p-ERK,p-JNK and p-p38 in the protein and mRNA level.Neiyi Zhitong decoction can prevent ectopic tissue pathological invasion and induce normal apoptosis by inhibiting activation of MAPK signaling pathway and increasing expression of caspase-9/caspase-3,which may be the mechanisms of treating EMs.