Effect Of Compound Rhizoma Curcumae Powder On Cell Apoptosis Of The Rat Models With Endometriosis

Objective:To observe the effect of compound Rhizoma Curcumae powder on endometriosis model in rat eutopic endometrium and ectopic endometrial histologic morphology and the effect on cell apoptosis, meanwhile, to observe the effect of compound Rhizoma Curcumae powder on apoptosis related gene surviving,caspase-3expression,to study the possible therapeutic mechanism of compound Rhizoma Curcumae powder from the cell apoptosis,to clatify the way of compound Rhizoma Curcumae powder on the treatment of endometriosis.Methods:70healthy female Wistar rats,body weight(200±20)g, the model of endometriosis(EMT) in Wistar rat was established by self-endometrium transplantation.Four weeks after the autotransplantation, the rats were paunched the second time to investigate the growth of transplantation and measure its volume. Those rats whose autotransplantation was not well growth were eliminated.The model rats with peritoneal implants were randomly assigned into four groups:the model control group and three treatment groups which were given respectively high-dose of compound Rhizoma Curcumae powder,low-dose of compound Rhizoma Curcumae powder and gestrinone served as the control drugs continuously for four weeks.Meanwhile,ten normal rats were used as the blank control. After4weeks treatment, after the last administration:in24hours abdominal anesthetized rats,we made the third laparotomy to measure the graft’s volume, compare the ectopic lesion volume for each group,and calculate the inhibition rate,strip the ectopic endometrium and the eutopic endometrium were stained with HE staining,to observe the pathomorphological changes with light microscopes, and TUNEL assay for detection of apopotosis in situ,real-time quantitative PCR assay for detection of ectopic endometrium in survivin,caspase-3gene expression.Results:1. The second laparotomy(4weeks after modeling)confirmed:under the method of establishing the model of EMT by self-endometrium transplantation without testing estrous cycle,achievement ratio was80.8%. The endometrial implants in the rats became cystic structures that were composed of endometrium,endometrial gland and stroma.Model4weeks before the assembly, compare the rat ectopic volume,the microscopic and histological appearance were consistent with endometriosis.No significant difference was found in the volume of grafts between model control group and each therapy group on the second laparotomy (P>0.05),treated for4weeks,a third laparotomy revealed in rats after administration of ectopic focus volume compared with pre-treatment,the difference was statistically significant(P<0.05); the ectopic lesion volume of the model control group at4weeks obviously increase, the difference was statistically significant(P<0.05),medication groups of ectopic lesion volume compared with the model control group, the difference was statistically significant(P<0.05).The medication group before and after treatment with inhibition rate:compared compound Rhizoma Curcumae powder in high dose group and the gestrinone group, the difference was not statistically significant(P>0.05),were higher than low-dose of compound Rhizoma Curcumae powder.2From the histological point of view, the ectopic endometrium of model control group growth is good, compound Rhizoma Curcumae powder in treating groups of lesions with atrophic changes.were similar with the gestrinone group,but had no significant effect on eutopic endometrium.3.TUNEL assay was used to investigate cell apoptotic index,we can found that,nomal rat endometrial cells in the presence have a certain cell apoptosis.the model control group, ectopic and eutopic endometrial tissue AI were significantly lower than those in blank control group, the difference was statistically significant(P<0.05),and the apoptosis of ectopic endometrial cells below the trate of eutopic endometrium,there was a significant difference(P<0.05),compound Rhizoma Curcumae powder of high,low dose group,the gestrinong group in eutopic endometrium.ectopic endometrium cells apoptosis rate were higher than those in the model control group,there were significant difference(P<0.05),the gestrinong group compared with low-dose compound Rhizoma Curcumae powder group,differences showed no statistical significance; high-dose compound Rhizoma Curcumae powder group of ectopic endometrial cells apoptosis rate obviously higher than that in eutopic endometrium.4.Real-time quantitative PCR detection of ectopic endometrium of apoptosis related gene survivin, caspase-3expression,we can found that,the model control group have high expression in ectopic endometrium of surviving and caspase-3low expression, high, low dose of compound Rhizoma Curcumae powder group were different degrees of up-regulation of caspase-3expression, at the same time the down-regulation of expression of survivin, difference has statistical significance (P<0.05). in high-dose of compound Rhizoma Curcumae powder group, the effect were especially significant.Conclusion:1. Compound Rhizoma Curcumae powder can inhibit the development of endometriosis, promoting ectopic focus atrophy and absorption, relieve abdominal adhesions, improving ectopic endometrial tissue pathological changes, while the eutopic endometrial tissue had no significant effect, in which the high-dose of compound Rhizoma Curcumae powder dose the most obvious.2.Compound Rhizoma Curcumae powder can reduced survivin in the rat ectopic endometrial tissue expression, improves the expression of caspase-3, thus enhancing the ectopic endometrial cells apoptosis ability, limited the ectopic endometrial cells proliferation and survival. So that the Compound Rhizoma Curcumae powder can inhibit the rat endometriosis development.and it’s mechanism might be closely associate with inhibiting the cell apotosis of ectopic endometrial.