The Biological Basis And Complications Of Phlegm Affected Baogong-2 Study Of Db/db Mouse Ovarian Dysfunction

Objective: Chinese medicine that phlegm type of infertility for Pathogenesis is " phlegm effect the baogong "- Mainly that is phlegm effect the baogong , phlegm that modern medicine is based on the biology of abnormal glucose and lipid metabolism. the db/db mouse with abnormal glucose and lipid metabolism, reproductive dysfunction, and other characteristics, With " phlegm effect the baogong " the symptoms of infertility, so we choose the db/db mouse based experimental studyExploration phlegm caused reproductive dysfunction in patients with the reasons to prove that the "phlegm effect the baogong" theory is correct. Ovarian also clearly within the leptin signal transduction pathway and ovarian partial loss of leptin signal ovarian function of their own.Method: Adult, female C57BL/6J mice (Jackson Laboratory, Bar Harbor, ME) were used in these studies. Littermate wide types (-/-), as well as diabetic (db/db) mutant genotypes, were matched for the allograft of the ovary. All mice were housed one per cage, under controlled environmental conditions (23C), with an established photoperi- od of 12 h light per day (lights on at 06:00 h). By ovary transplantation on the day of 12 weeks, five genotypic mice were constructed with body LR+ and ovary LR- in Group A, body LR+ and ovary LR- in Group B, body LR+ and ovary LR+ in Group C, body LR- and ovary LR+ in Group D, as well as body LR-and ovary LR- in Group E respectively, each mice bearing only one ovary in the experiment. the vaginal smear was done to evaluate the recovery and cyclicity of transplanted ovaries with mutant or intact LR. Before execution, all mice were injected leptin (1h before), and then the ovaries were collected for the detection of the expression of leptin ,ob-Rb,JAK2,stat3 , p-stat3,socs3, and the PIAS3 by the way of the immunohistochemistry, and the RT-PCR was performed to examine the mRNA expression of ob-Rb, SOCS3,and the serum were reserved for the assay of the glucose, insulin, LDL-c, E2, P, FSH and LH. The weight of each mouse, their ovary and their fat pads were also recorded.Results: Mice in D and E groups showed the similarly profiles of the glucose and fat metabolism which were hyperglycemia, hyperinsulinemia and hyperlipidemia, and were significantly higher than those of A, B and C groups, although they presented comparably normal levels. Moreover, mice with mutant body LR in D and E groups were acyclic (i.e., persistent diestrus) and anovulatory (i.e., lack of ovarian luteinization) corresponding to diminished steroidogenic capacity and gonadotropin levels, although ovaries in two goups are genetipically different in LR. Mice with intact body LR in A, B, and C groups demonstrated normal cycles with similar gonadotropin and steroid levels each group bear ovary with mutant or intact LR. The RT-PCR analysis showed ,after significant higher leptin activated, the expression of ob-Rb in ovaries of C and D groups,none of the expression of ob-Rb in ovaries of A ,B and E groups mice, and socs3 higher expression in LR+ ovaries in C group as compared with the other groups. Immunohistochemistry showed that leptin protein was higher expressioin in ovaries of A ,B and E groups than in ovaries of C and D groups,and ob -Rb protein was expressioin in ovaries of C and D groups,unexpression in ovaries of A ,B and E groups mice,and jak2 protein was expressioin in ovaries of C and D groups,unexpression in ovaries of A ,B and E groups mice,and stat3 protein was expressioin in ovaries of A,B,C,D and E groups,and p-stat3 protein was expressioin in ovaries of C and D groups,unexpression in ovaries of A ,B and E groups mice,and socs3 protein was expressioin in ovaries of A,B,C,D and E groups, and PIAS3 protein was expressioin in ovaries of A,B,C,D and E groups.Conclusion: Ovarian to exist integrity of the leptin signal transduction systems, and ovarian leptin signaling deficiency does not affect ovarian their own reproductive and endocrine function; db mouse ovarian dysfunction is high glycolipids hyperlipidemia involving genital lead to induced lipid.Clearly, " phlegm effect the baogong " can cause infertility clinical phlegm - reproductive dysfunction in patients with this theory is correct, and abnormal glucose and lipid metabolism is "phlegm effect the baogong" the biological basis.