Effect Of Huluan Decoction (HD) On Expression Of CD4~+ CD25~+ Regulatory Tcells And FoxP3 In The Immunological Premature Ovarian Failure Of Model Mice

Objects:To initially explore the effect and mechanism of Chinese Medicine on POF. The mechanism of protective effect of HD on immunological POF was studied.Methods:1. The knowledge, treatment and research status of POF were reviewed from TCM and western medicine. The effect and mechanism of CD4+CD25+ regulatory T cells and FoxP3 in autoimmune diseases were introduced. The experimental research plan about the effective mechanism of HD on immunological ovary premature senility was set.2.The immunological ovary premature model was set by immunogens of 330th-342th amino acid sequences (NSSSSQFQIHGPR) and HD was used to treat the model. The morphology of ovary, anti zona pellucida antibody and the expressive changes of CD4+CD25+regulatory t cells and FoxP3 were detected by optical microscope, enzyme-linked immunoassay, flow cytometry, reversed transcription polymerase chain and reaction(RT-PCR) and Western-Blot.Results:1.In model group:Estrous cycle was disorder gradually, the time was relative earlier. The vaginal cell smear showed that there were a large number of leukocytes and a few of nucleate epithelial cells.the volume of ovary was reduced. The mature follicle was less. The granular cell layer of follicle was less than normal and arranged in disorder. Some follicles were dysplastic. There were much more lymphocyte and plasma cell infiltration.AzpAb in peripheral blood was positive and the level was increased.The indexes of ovarian weight and thymus weight were decreased, while the weight of spleen was increased.The number of CD4+CD25+ regulatory t cells was decreased.The expression of FoxP3 gene and protein was less.2.In HD group:The above status of POF mice were improved in different degrees. The effect of high dose group was obvious than that of middle dose group which was showed as following:Estrous cycle disorder appeared later than that in model group which showed the estrous cycle was prolonged, the keratinocyte in estrus was decreased. The estrous cycle was prolonged 8-10 days. Morphology of ovary: Compared with model group, the volume of ovary increased and there were more mature follicles. The granular cell layer of follicle was increased. A small part of follicles was dysplasia. There were lymphocyte and plasma cell infiltration in ovarian stroma. AzpAb in peripheral blood was positive while the level was decreased obviously.The indexes of ovarian weight and thymus weight were increased obviously, while the weight of spleen was decreased remarkably.The number of CD4+CD25+ regulatory t cells was increased.The expression of FoxP3 gene and protein was increased.Conclusion:The main etiology and pathogenesis of POF is due to deficiency of kidney and pregnant abdominal pain. Reinforcing kidney and dredging collaterals is the basic principle.The model is close to human beings and is one of the ideal models.HD has some improvement effects on the function of ovary and immune of mice with immunological ovary premature senility.