| Premature ovarian failure(POF) is a disease that loss of function of the ovaries before age 40 and defined by abnormally low level of estrogen and high level of FSH. It was reported that Guishen Pill was used in the treatment of POF and menstruation diseases. It should provide theoretical basis and help build diagnostic criteria through study the treating effort and mechanism of Guishen Pill on POF. In this study, we evaluated the treating effort of extraction of Guishen Pill on apoptosis of immune POF mice and hope to find out the mechanism of Guishen Pill on POF.70 SPF BALB/c mice was randomly divided into blank control group(n=10) and model group(n=60). The mice of model group were induced by subcutaneous multipoint injection with pellucid zone ZP3 peptide(ZP3) No.330~342 animo acid sequence of mouse in complete Freund's adjuvant(CFA) as immunogen to establish POF animal model while mice of blank control group were injected with saline solution. The model animal were randomly divided into model control group, Estradiol estradiol group Guishen Pill lowdosage, middle-dosage and high-dosage group(n=12) according to the body weight of mice. And the mice of each group were treated with drugs accordingly for 4 weeks and then all mice were killed and ovary, womb, spleen, thymus and hypophysis were collected and weighted. Serum of each mouse was collected and interleukin-17(IL-17) was detected. Number of follicles, corpora luteum and interleukin-1? were observed and calculated based on histophysiology and immunohistochemical stains.The results shoued that: The estrous cycle of model mice became irregular compared with blank control group. The secondary follicles, mature follicles and corpora luteum decreased significantly while corpora atretica increased compared with blank control group. The weight indices of ovary, womb, thymus and hypophysis increased while weight of spleen descended. After treatment that last for 4 weeks, the number of secondary follicles of the mice of Estradiol estradiol group and Guishen Pill low-dosage, middle-dosage and high-dosage groups increased significantly compared with model control group(p<0.05?p<0.05?p<0.01?p<0.05). Mature follicles of the mice of Estradiol estradiol group and Guishen Pill middle-dosage and high-dosage groups increased significantly compared with model control group(p<0.05). The number of corpora atretica of the mice of Estradiol estradiol group and Guishen Pill low-dosage, middle-dosage and high-dosage groups decreased significantly compared with model control group(p<0.05). And the number of corpora luteum of Guishen Pill high-dosage group increased significantly compared with model control group(p<0.05).The expression of IL-1? of ovarian follicular fluid in oocytes of the mice of Estradiol estradiol group and Guishen Pill low-dosage, middle-dosage and high-dosage groups decreased significantly compared with model control group(p<0.01), while no significantly differences of the expression of IL-1? was found in cumulus cells(p>0.05). The expression of IL-17 in the serum of the mice of Estradiol estradiol group and Guishen Pill low-dosage, middle-dosage and high-dosage groups decreased significantly compared with model control group(p<0.01?p<0.05?p<0.05?p<0.05). The results showed that the expression of IL-17 and IL-1? of model control group mice increase significantly and the extraction of Guishen Pill could increase the number of secondary, mature follicles and corpora luteum while decrease the number of corpora atretica. The effect of reducing IL-17 and IL-1? was dosage-depended.POF model mice were induced by subcutaneous multipoint injection with pellucid zone ZP3 peptide(ZP3) No.330~342 animo acid sequence of mouse in complete Freund's adjuvant(CFA) as immunogen in this study. And estrous cycles irregular and high expression of IL-1? in ovaries of mice indicated that animal model was established successfully. Guishen Pill can adjust autoimmune disorder through prohibiting the expression of IL-17 and IL-1?, thus improve the function of ovary. |
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