Study Of The Effect And Mechanism Of "ping Chong Jiang Ni Formula" On The Migration And Invasion Of Ectopic Endometrial Stromal Cells

Objective :To investigate the effect and mechanism of "Ping chong Jiang ni formula" on EMS from the cellular molecular and proteomic level by establish the Ectopic endometrial stromal cells(EESCs)model of isolated human endometriosis.Methods:The interstitial cells of EMS were digested,cultured and identified in vitro by using collagenase digestion of ectopic lesion tissue,sieve filtration tissue digestion,centrifugal separation and cell Passaged and establish EMS ectopic interstitial cell model."Ping chong Jiang ni formula"(the“ZY”)and normaol saline(NS)lml / 100 g were given to the SD rats twice daily,and serum was extracted after 14 days of administration.Then the interstitial cells were cultured with 10% FBS(blank serum group),10% NS serum,5% ZYserum,10% ZY serum and 15% ZY serum for 24 h,48h and 72 h respectively.observing the number of cells of the migration and invasion by Transwell chamber method.the interstitial cells were cultured in ZY group and NS group for proteomic analysis of TMT technique.Through literature review and analysis,found out the differentially expressed proteins which are significantly related to migration and invasion,and discussed their possible key targets.Results:1.EESCs were successfully isolated and cultured,and cell model of isolated human endometriosis was constructed.2.Comparing the migration and invasion ability of EESCs in EMS patients treated with five different treatment methods for 24 h,48 h and 72 h,It was found that the number of migration and invasion cells of different serum concentrations and time in ZY group was less than that in FBS group and NS group,the difference was significant(P < 0.05).Besides,The number of in groups containing 5% ZY serum was less than that in groups containing 10% and15% at each time,the difference was significant(P < 0.05).but there was no significant difference in groups containing 10% and 15% ZY serum(P>0.05).The number of cells treated with ZY serum at 48 h was significantly higher than that at 24h(P < 0.05),but there was no significant difference between 48 h and 72h(P>0.05).The results indicated that "Ping chong Jiang ni formula" could inhibit the migration and invasion of EESCs,and 48 h is the best time,10%ZY is Optimum concentration.3.A total of 5048 proteins were identified in human by mass spectrometry and database retrieval using TMT technique.Among the proteins with significant differences in ZY group VS NS group,3 differentially proteins were up-regulated and 9 differentially proteins were down-regulated.Conclusion:1.It laid a experimental research foundation for studing the molecular mechanism of "Ping chong Jiang ni formula" on EESCs that the successful preparation of "Ping chong Jiang ni formula" containing serum and establishment of EMS ectopic stromal cell model in vitro.2."Ping chong Jiang ni formula" can inhibit the migration and invasion of EESCs cultured in EMS in vitro.3.The inhibition of "Ping chong Jiang ni formula" on the migration and invasion of EESCS cultured in EMS in vitro may be related to the proteins Fer and TRIP13.It may act by regulating adhesion and EMT.In order to provide a more comprehensive theoretical basis for the mechanism of action of "Ping chong Jiang ni formula" in the treatment of EMS,the specific association still needs to be verified by proteins and further discussion on the mechanism of action.