The Screening Of Key Genes And Mechanism Associated With Inflammation That Acted By Neiyi Zhitong Decoction On EMs Rats Model

Objective:To screen the differential expression genes in SD rats endometriosis model treated by Neiyi Zhitong decoction via whole genome expression chips,we explore the molecular mechanism of NZD in the treatment of endometriosis. Based on the chip results, we detected the estradiol, TNF-a,COX-2nserum,the expression of NF-KBp65 and VEGF n ectopic endometrium tissue,to investigate the mechanism of NZD on the effect of anti inflame-matory and anti angiogenesis, in order to provide a comprehensive evaluation of NZD’S therapeutic effets and to offer the experimental basis for the research and development of new drugs.Method:A homologous rats endometriosis model was established by transplanting the uterine tissue into the abdominal cavities of recipient rats.4 weeks later, the successful model of the 32 SD rats were randomly divided into 4 groups with 8 rats in each group,the model group, Neiyi Zhitong decoction low dose group,NZD high dose group, the Danazol group,as well as 8 rats in the sham operation group.Except the sham operation group,they were given sterile water, NZD decoction in low dose group (1.33g/kg),NZD decoction in high dose group(5.32g/kg) and Danazol(7.2mg/d) orally for 4 weeks.After the last administration 24 hours later, The endometriosis tissue, peritoneal fluid, serum were collected.Avoiding any form of contamination and the tissue was properly processed and stored.Tissue from Neiyi Zhitong decoction’ low dose group and the model group were detected by whole genome expression chips,the level NF-κBp65 and VEGF were verified by QT-PCR.Serum Estradiol and progesterone were detected using radioimmunoassay.ELISA methods was used to detect the serum and peritoneal fluid levels of TNF-a COX-2. Immune histochemical method for the detection of ectopic endometrial tissue NF-κBp65 protein.Results:1.Compared with model group, Neiyi Zhitong decoction and Danazol treated rats showed significantly reduced endometriotic implant volumes(P<0.05).2. we discovered 19 differentially expressed genes,which include:Ngfr, Pgr,Prlhr,Ghrhr, Agtr2, Ccl3, Grik1, Comt, Kcnip3, Fgfr4, Foxa2, Cc17, Cd40, Apob, Pf4,Mmp12, Ccnbl, Vegfa by the analysis from whole genome expression chip.3. E2 levels in serum was decreased in NZD low group(P<0.05),which levels were significantly higher in control group,the serum progesterone did not significant changed.4. A significant and dose-dependent decrease n activities of TNF-α,E2 and COX-2 in serum and peritoneal fluid of the rats in lower dose NZD Group and higher dose which NZD Group was detected by ELISA(P<0.05).5.Similarly,tissue levels of NF-κBp65,VEGF mRNA and p65 protien were significantly lower in NZD group than that of control animals.Conclusion:1. NZD decoction showed a meliorative effects on endometriotic implants,improve the morphological changes, in a dose dependent manner.2.By gene chip technology, we screened 19 genes related to NZD decoction on the treatment of EMS rats.3.Neiyi Zhitong decoction can reduce the level of serum E2, decrease the expression of TNF- alpha and COX-2 in serum and peritoneal fluid.4.Neiyi Zhitong decoction could decrease the mRNA and protein expression of NF-κBp65 in ectopic tissues in EMs rats model.5.Targeting inflammation, estrogen production and angiog-enesis with Neiyizhitong decoction therapy limited the development of invasion and angiogenesis associated with ectopic endometrial growth.