Effects Of Cuscuta Flavonoids And Ban’s Ovum-Stimulating Soup On The Maturation Of Mouse Oocytes

Objective: To investigate the effects of Cuscuta flavonoids on mouse oocytes in vitro culture and Ban’s ovulation-stimulating soup on the maturation of mouse oocytes with polycystic ovary syndrome,to explore Cuscuta flavonoids and the use of Cuscuta as a prime drug The mechanism of the compound on the in vitro maturation of mouse oocytes.Method: 1.Isolate mouse oocytes for in vitro culture,add Cuscuta flavonoids at concentrations of 10ul/m L,5ul/m L,1ul/m L,and observe the Cuscuta flavonoids by histopathology and immunofluorescence staining techniques.(SCF)influence on the level of intracellular reactive oxygen species(ROS),the influence of intracellular ATP content,and the influence on the ultrastructure of oocyte cytoskeleton.Western-blotting was used to detect the protein expression of GDF9 and BMP15,and q-PCR was used.Detect the gene expression of GDF9 and BMP15;2.Use 21-day-old KM mice to subcutaneously inject DHEA 9mg/100 g combined with medical soybean oil(0.1m L/100g)on ??the back of the neck every night,once a day,for 21 consecutive days,A mouse model of PCOS was prepared and randomly divided into 5 groups,the model control group,the Diane-35 group(0.035mg/kg),the high,medium,and low-dose groups of Ban’s Ovum-stimulating Decoction(2g/ml,1g/ml),0.5g/ml).The blank control group and the model control group were given the same volume of normal saline every day,and the other groups were given corresponding drugs by gavage.For 25 consecutive days,the blood was collected from the eyeball and the serum was separated for the determination of FSH,LH,PRL in the serum.,PRO,E2,and T content.The ovarian oocytes of each group of mice were isolated and cultured in vitro.Western-blotting was used to detect the protein expression of GDF9 and BMP15,and q-PCR was used to detect the gene expression of GDF9 and BMP15.Results:1.Experimental results:(1)Compared with the blank control group,the middle concentration group can reduce the apoptosis of oocytes(P<0.05).(2)Compared with the blank control group,different concentrations can reduce the oxidative stress of oocytes and increase the activity of oocytes(P<0.01).(3)Compared with the blank control group,the medium concentration and the high concentration can effectively improve the abnormal morphology of the cytoskeleton and improve the cell quality(P<0.01).(4)Compared with the blank group,both low concentration and high concentration can increase the relative expression2.of BMP15 m RNA in oocytes(P<0.05).(5)Compared with the blank group,all three concentrations can increase the relative expression of GDF9 m RNA in oocytes(P<0.05).(6)Compared with the blank group,both medium and high concentrations can increase the relative expression of BMP15 protein in oocytes(P<0.01).(7)Compared with the blank group,all three concentrations can increase the relative expression of GDF9 protein in oocytes(P<0.01).(8)Compared with the blank group,the medium concentration group can significantly increase the intracellular ATP content(P<0.01).2.Experimental results:(1)DHEA induced a successful mouse PCOS model.Compared with the blank control group,the ovarian tissue of the model control group was abnormal;compared with the blank control group,the testosterone(T)of the model control group was higher than that of the blank group(P<0.01)(2)Compared with the blank control group,the serum levels of T,E2,LH,and PRL in the model group increased significantly(P<0.01),the P level decreased significantly(P<0.01),and the FSH level decreased significantly(P <0.05);(3)Compared with the model group,the Diane-35 group and the high,medium,and low-dose Chinese medicine compound group made the ovarian tissue of PCOS mice basically return to normal,which can reduce the serum T,E2,LH,and PRL Decreasing the level(P<0.01)can significantly increase the level of P in the serum(P<0.01),and large and medium doses can significantly increase the serum FSH level in PCOS rats(P<0.05);(4)and the model control group In comparison,the content of GDF9 and BMP15 m RNA and protein in the Diane-35 group and the high,medium,and low dose groups increased(P<0.01);Conclusion: 1.Cuscuta flavonoids can reduce the oxidative stress of oocytes,reduce cell apoptosis,improve the abnormal arrangement of cytoskeleton,increase the content of ATP in cells,and increase the content of GDF9 and BMP15,which are related to oocyte maturation..To achieve the purpose of improving the maturation rate of oocytes.2.The compound Ban’s Ovum-stimulating Soup with Cuscuta as the master drug can regulate the hormone levels of the mouse PCOS model and promote the maturation of oocytes.The mechanism is also by increasing the content of oocyte maturation-related proteins GDF9 and BMP15.It shows that Ban’s Ovum-stimulating Soup with Cuscuta as the prime medicine can promote the maturation of oocytes.