Effects Of Serum Contained ZiLe Tiao Jing Soup On The Proliferation And Apoptosis Of Ovarian Granulosa Cells From SD Rats

Objective:By observing the effects of the different concentrations of serum contained Zi Le TiaoJing Soup?ZLTJS?on proliferation and secretion of estradiol?E₂?and progesterone?P?,activin A?Act A?of ovarian granulosa cells?GC?cultured in vitro form SD rats,to explore the the mechanism of action of ZLTJS and provide the theoretical basis for further popularization and application of ZLTJS in clinic.Methods:1.Using ZTLJS with a certain concentration and normal saline to lavage nearly mature 6-week-old healthy SD rats,to preparate the serum contained ZLTJS and serum contained no medicine of rats,And configure into three groups?small,medium and large?of different concentrations of serum contained ZLTJS and serum contaied no medicine with the volume fraction of 10%,And set up the blank control group;2.The method uses a simple mechanical separation and trypsin digestion to preparate ovarian granulosa cells?GC?with 3 weeks of healthy SD rats,and set up a in vitro cultivation system,observe the growth of rat ovarian granulosa cells under an optical microscope,through FSHR immunohistochemical method and cell climb HE staining method to identify the cells,Draw the cell growth curve by MTT;3.Based on the principle of the method of serum pharmacology,after 24 h when GC cultured in vitro is adherent,use a small,medium and large different concentrations of serum contained ZLTJS and serum contained nomedicine of rats to intervente the ovarian granulosa cell,and detect absorbance value by MTT to observe granulosa cell proliferation;4.Detect the content of ActA in the medium by radiation immunity analysis method;5.Detect the content of ActA in the medium by ELISA.Results:1.The ovarian granulosa cells cultured in vitro grew well,24 h almost adherent,3-4 d enterd the proliferation stage,about 15 day cell apoptosis significantly,FHSR immunohistochemical staining:The nucleus was blue purple,cytoplasm was brown yellow,the positive rate of more than90%;HE staining:Nuclei was dark blue,cytoplasm was pink;2.The effect of serum contained ZLTJS on ovarian granulosa cells?GC?proliferation of rats:the OD values of the small,medium and junior groups of different concentrations of serum contained ZLTJS were significantly higher than and the serum contained no medicine group and blank control group,The difference was statistical significance?P <0.01?,OD in each group of serum contained ZLTJS has a certain dependence with concentration,and the the differences between groups were statistically significant?P <0.05?;3.The effect of serum contained ZLTJS on ovarian granulosa cells secretion of rats:The secretion of E₂,P,ActA of the small,medium and junior groups of different concentrations of serum contained ZLTJS were significantly higher than and the serum contained no medicine group and blank control group,The difference was statistical significance?P <0.01?,The secretion of E₂,P,Act A in each group of serum contained ZLTJS has a certain dependence with concentration,and the the differences between groupswere statistically significant?P <0.05?.Conclusions:Serum contained ZLTJS can promote granulosa cell of rats proliferation and promote the secretion of E₂,P,ActA,.It is speculated that in the body,serum contained ZLTJS may work by improving the ovarian local autocrine and paracrine and raising Act A secretion affect Act A/Smads signal pathways,So as to regulate ovarian local microenvironment and achieve the purpose of adjusting the ovarian function.