Effect Of Zuogui Wan On Three Dimensional Model Of Angiogenesis And Its Relationship With Expression Of PLGF And Flt-1 In Ovary

Objective: To investigate the effect of angiogenesis in vitro three-di-mensional model by serum of Zuogui Wan under HPLC monitoring and the expression of PLGF and Flt-1 in the ovaries of rats with diminished ovarian reserve by Zuogui Wan.To explore the mechanism of the function of Zuogui Wan in promoting angiogenesis.Method:(1)45 famale Sprague-Dawley rats weight for 220 ± 20 g were randomly divided into 3 groups,the rats were treated with distilled water(20g/kg),Zuogui Wan dose(11g/kg)and Zuogui Wan dose(33g/kg),respectively.Then Zuogui Wan serum was prepared in different time points.Select the Zuogui Wan serum of largestpharmaceutical ingredient peak time to three-dimensional HUVEC culture in vitro.MTT was used to observe Zuoguiwan containing serum on HUVEC proliferation.Vessel surface culture method of rat tail collagen type I was used to establishment of HUVEC three-dimensional model.ELISA detection of estradiol andvascular endothelial growth factor levels.(2)40 famale Sprague-Dawley rats weight for 180±20g were divided into 8 of normal group and 32 of model group,tripterygium glycosides gavage(50mg/kg)was used for 14 days lavage to establish diminished ovarian reserve rat model,Successful models were treated with distilled water(20g/kg),Zuogui wan low dose(11g/kg),Zuogui wan high dose(33g/kg),conjugated estrogen(65pg/kg),respectively.The estrous cycle were observed by vaginal smear.The ovary and uterus indexes were calculated.Ovarian morphology changes were observe by HE staining.The levels of PLGF and itsreceptor Flt-1 in the rat ovaries were determined by immunohistochemistry and Western blot in each group.Results:(1)Two hours after administrated Zuoguiwan was the highest peak drug absorption.Zuoguiwan containing serum low dose group(72h),high dose group(24,48,72h)were proliferated HUVEC(P<0.05 or P<0.01).Under three-dimensional culture,Zuoguiwan low and high dose group may increase endothelial tubules area,length,diameter and generate index(P<0.01),Zuoguiwan group could improve serum E2,VEGF levels(P<0.05 or P<0.01).(2)The estrous cycle extended in the diminished ovarian reserve rats,and gradually returned to normal after treatment.The ovary and uterus indexes were higher in rats receiving treatment than in the model group(P<0.05 or P<0.01).The number of follicles,corpus luteum and ovarian vessels increased(P<0.05 or P<0.01),and the number ofatresia follicles decreased after treatment(P<0.05 or P<0.01).Integrated optical density and the levels of PLGF and Flt-1 increased in the treatment group(P<0.01).Conclusion:(1)Zuogui Wan can promote the proliferation and of vascular endothelial cells,improve the structure of three-dimensional tube monitor by HPLC.(2)Zuogui Wan can gradually restore emotional cycle of Tripterygium wilfordii polyglycoside rats,increase ovarian and uterine index,improve ovarian tissue morphology and Number of ovarian vessels.(3)Zuogui Wan can reducing E2 and VEGF levels in serum and up-regulating the expression of PLGF/Flt-1 in ovarian tissue,then promote the formation of ovarian blood vessels Thereby improve ovarian reserve function.