ABSTRACT
Objective:The effects of anemoside B4 on endometritis rats were studied through in vivo and in vitro experiments. Method:Animal experiments used 25% phenol glue to prepare endometritis models. 60 female SD rats were randomly divided into blank group， model group， Kushen gel group（0.005 g·kg-1），anemoside B4 gel low，medium and high dose groups（0.005，0.01，0.02 g·kg-1），10 rats in each group，except for the blank group，rats in each group were injected with 25% phenol glue into their vagina every 2 days，and the modeling continued for 30 days. Administration started on the day after modeling. Anemoside B4 gel low， medium and high dose groups were administered rectal daily，Kushen gel group was given daily vaginal administration. The blank group and model group were given the same amount of normal saline in the same way for 30 consecutive days. After the last administration，the uterus and its attachments of each group of rats were taken to analyze the uterine morphology and index. Hematoxylin-eosin （HE） staining was used to observe the pathological changes of rat uterus. Real-time PCR was used to detect tumor necrosis factor-α （TNF-α），interleukin-1β （IL-1β），and interleukin-6 （IL-6），signal transduction protein 130 （gp130），signal transduction and transcription activator 3 （STAT3）mRNA expression. Detection of IL-6 and STAT3 protein expression in rat uterus by Western blot. In cell experiments，lipopolysaccharide （LPS）was used to induce rat endometrial epithelial cells to prepare an in vitro inflammation model， and Real-time PCR was used to detect the expression of IL-6，gp130 and STAT3 mRNA in each group of rat endometrial epithelial cells. Result:The results of animal experiments showed that compared with the blank group， the model group had inadequate uterine cavity adhesions， endometrial edema and hyperemia. Compared with model group， there was no adhesion in the uterine cavity of the rats in the high dose anemoside B4 gel group and the Kushen gel group. The uterine tissue was relatively complete， and the uterine pathological structure was significantly improved. Compared with the blank group， the uterine index of the model group was significantly increased（P<0.05）， the expression of IL-1β mRNA in the uterine tissue was significantly increased （P<0.05）， the expression of mRNA and protein of IL-6 and STAT3 in the uterine tissue significantly increased （P<0.05）. Compared with model group， the uterine index in anemoside B4 gel high dose group was significantly reduced （P<0.05）， and the mRNA and protein expression levels of IL-6 and STAT3 in the uterine tissue were significantly reduced （P<0.05）. There was no statistically significant difference between the TNF-α and IL-1β mRNA expression compared with the model group. Cell experiment results showed that compared with the blank group， the mRNA expression of IL-6 and gp130 in model group endometrial epithelial cells was significantly increased （P<0.01）， STAT3 mRNA expression was significantly increased （P<0.05）. Compared with model group， the mRNA expression levels of IL-6， gp130 and STAT3 in anemoside B4 high dose group decreased significantly （P<0.05）. Conclusion:Anemoside B4 can improve the inflammatory response of chronic endometritis in rats and reduce the release of inflammatory factor IL-6. The mechanism may be related to the down-regulation of IL-6/STAT3 pathway.
ABSTRACT
Objective:To detect expression of tumor necrosis factor-converting enzyme(TACE)on the mouse uterine epithelial cell and effect of implantation in vitro. To analyze its role in blastocyst implantation and to investigate the relation TACE withimplantation of mouse embryo. Methods:Co-culturing endometrial epithelial cells and blastocyst of the pregnant 4-day mouse to establish the implantation model in vitro. Epithelial cells was identified by Immunocytochemistry. RT-PCR and Immunocytochemistry were used to detect the expression of TACE gene and protein in the endometrial epithelial cells. Different concentrations of TACE antibody were added to the implantation model to observe effect of TACE antibody on mouse blastocyst implantation in vitro. Expression of TACE protein was detected by Immunocytochemistry in each antibody interference system. Results:Endometrial epithelial cells growed vigorously in vitro. Keratin antibody showed positive expression,meanwhile,Vimentin antibody showed negative expression in the endometrial epithelial cells. PCR showed that the expression of TACE gene was high and results of Immunocytochemistry was consisted with PCR. The adhesion rate of mouse blastocyst was high in the co-culture system and decreased remarkably when TACE antibody was added (P
ABSTRACT
Objective To study the biological effects of four isoflavone derivatives(F8,F11,ZF3 and ZF7)on endometrial epithelial cells.Methods The endometrial epithelial cells were cultured through collagenase enzymatic digestion and twice grit filtration.The biological effects on endometrial epithelial cells of four isoflavone derivatives were compared through MTT.Results The primary endometrial epithelial cells were successfully disassociated,cultured and passaged down stably.Cell proliferation was significantly increased by F8(25 mol/L)(P