The Effects Of Bushenhuoxue Medicine-contained Serum Decoction On Cultured Autoimmune Premature Ovarian Failure Mice Regulatory T Cells

Objective:By in vitro culture autoimmune ovarian premature senility of regulatory T cells in mice, observed bushenhuoxue medicine-contained serum effect on CD4+CD25+Treg proliferation.Methods:Hybrid breeding in A/J males and C57BL/6 females,we succeeded in gettingjian the Healthy B6AF1 mice. Screening of sexual cycle 30 normal female mice into 2 groups at random:model group (20 mice),normal group (20 mice). Model group by the tail and left foot after subcutaneous injection of immune reagent building, each mouse multi-point injection volume of 0.1ml (a mixture of pZP3+CFA). at weeks 2 after establishing models,6 mice were selected randomly from the normal group and model group to evaluate the mice model. Take blood by removalling eyeball, estradiol level change detected by ELISA assay in mouse serum, then take the thymus, spleen, uterus and ovaries, weighing. vaginal exfoliated cells stained by Papanicolaou, ovarian tissue morphology observed by H&E staining and ZP3 expression in ovarian tissue examined by immunofluorescence staining were together used to judge if the animal model is available. Screening 8 Wistar mice into 4 groups at random:Blank group bushenhuoxue group of high, medium and low dose group (2 mice each). At the same time, the mice were began to administrated orally for continuous 1 week, medicine group mice were administered 31.5 g/kg Bushenhuoxue Recipe (20g/kg、10g/kg、5g/kg), blank group mice were administrated orally the equal volume of double distilled water. Blood samples were obtained from abdominal aorta after 1 week, To obtain the drug-containing serum. To isolate the CD4+CD25+Treg by Magnetic activated cell sorting, Flow cytometry test purity, and culture it With drug-containing serum.Detection of CD4+CD25+Treg proliferation by CCK-8 and CFSE.Results:After building, Viscera index in mice, Estradiol level, and the ovarian tissue of model mice were used to evaluate the model of Immunological Premature Ovarian Failure. All experimental group Can proliferate the CD4+CD25+Treg, Medicated serum group higher than PBS group obviously;the proliferative effect of middle Medicated serum group are better than low and high group, P<0.05, the difference was statistically significant.Conclusion:Bushenhuoxue medicine-contained serum has obvious promoting effecton proliferating of CD4+CD25+Treg.