| Objective:By investigating the influence of Neiyi Zhitong Decoction on the histopathology and the expression of Bcl-2,Bax,p-JAK2,p-STAT3,SOCS3 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.At the same time,we measured the expression of Bcl-2 and Bax,p-JAK2,p-STAT3 and SOCS3 protein of intrimas by using the immunohistochemistry and Western Blot method,And the expression of p-JAK2,p-STAT3 and SOCS3 mRNA was measured by RT-PCR method.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 Bcl-2,Bax protein,compared with sham operation group,the expression of Bax protein of model group increased and the expression of Bcl-2 protein decreased(P<0.05);compared with the model group,the expression of Bcl-2 protein in the low group and high group and Western medicine group decreased and Bax protein increased(P<0.05);comparing that in the low group,high group and Western medicine group,there is no statistical significance(P>0.05).4,On the view of the expression of p-JAK2,p-STAT3 and SOCS3 protein,the expression of p-JAK2 and p-STAT3 protein in model group was higher than that of sham group but SOCS3 protein expression was lower(P<0.05);compared with the model group,the expression of p-JAK2 and p-STAT3 protein in high group and western danazol group decreased,but SOCS3 protein increased(P<0.05);compared with the low group,the expression of p-JAK2 and p-STAT3 protein in high group and western danazol group decreased,but SOCS3 protein increased(P<0.05).5,In JAK2,STAT3,SOCS3 mRNA expression,compared with the sham operation group,the expression of JAK2 and STAT3 mRNA in the model group was increased,and the expression of SOCS3 mRNA was decreased(P<0.05);compared with the model group,the expression of JAK2 and STAT3 in the low,high group and Western medicine group decreased,and the expression of SOCS3 mRNA increased(P<0.05);compared with the low group,the expression of p-JAK2 and p-STAT3 mRNA in high group and western danazol group decreased,but SOCS3 mRNA increased(P<0.05).Conclusion:1,Neiyi Zhitong decoction can reduce endometriotic implants and improve the morphological changes.2,The unbalance of apoptosis factor Bcl-2/Bax and the over activation of JAK2/STAT3 signaling pathway play an important role in the pathogenesis of EMs.3,Neiyi Zhitong decoction can reduce the Bcl-2 protein and increase Bax protein,so it reduce the ectopic focus by promoting ectopic cell apoptosis and inhibiting the proliferation of ectopic tissue.4,Neiyi Zhitong decoction can restore the normal expression of SOCS3 and inhibit the pathological activation of JAK2/STAT3 signaling pathway by reducing the expression of p-JAK2,p-STAT3 and increasing the expression of SOCS3 in the protein and mRNA level.5,Neiyi Zhitong decoction can prevent ectopic tissue pathological invasion and induce normal apoptosis by inhibiting activation of JAK2/STAT3 signaling pathway and balancing expression of Bcl-2/Bax,which may be the mechanisms of treating EMs. |
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