Effect Of Xihuang Pill On Hypothalamus-Pituitary-Ovarian Axis In Rats

Objective:To study the effect of Xihuang Pill on hypothalamic-pituitary-ovarian axis related hormones,receptors and ultrastructure under transmission electron micr-oscope in rats,and to clarify the intervention effect and mechanism of Xihuang Pill on hypothalamic-pituitary-ovarian axis in rats,so as to provide a theoretical basis for clin-ical use of Xihuang Pill to prevent and treat hypothalamic-pituitary-ovarian axis dys-function.Methods:Wistar female non-pregnant rats were divided into 5 groups according to random number table method,with 10 rats in each group:blank group,tamoxifen group,Xihuang pill high,medium and low dose group.Tamoxifen group was given 1.8mg?kg^-1?d^-11 tamoxifen by gavage,while high,medium and low dose groups were given2.16,1.08 and 0.54g?kg^-1?d^-11 Xihuang pills by gavage,while the blank group was giv-en equal volume distilled water,with the volume of each group being 3mL.After the l-ast administration,the indexes of ovarian and uterine organs were calculated,and the l-evels of gonadotropin releasing hormone,follicle stimulating hormone,luteinizing hor-mone,estradiol and progestogen in serum,follicle stimulating hormone and luteinizing hormone in pituitary,estradiol and progestogen in ovary were detected by enzyme-linked immunosorbent assay.Gonadotropin-releasing hormone receptor in pituitary,follicle stimulating hormone receptor and luteinizing hormone receptor in ovary were detected by Western blot.The expression of gonadotropin-releasing hormone receptor,follicle stimulating hormone and luteinizing hormone genes in pituitary,follicle stimu-lating hormone receptor and luteinizing hormone receptor genes in ovary were detec-ted by Real-time fluorescence quantitative PCR.Immunohistochemical method was used to detect estradiol receptor?,estradiol receptor?and progestogen receptor in ovary.HE staining was used to observe the morphology of ovarian tissue cells.Ultra-structure of pituitary and ovarian tissues was observed by transmission electron mic-roscopy.Results:1.From the level of serum hormone,compared with the blank group,the level of serum gonadotropin releasing hormone in the high and middle dose groups of Xihuang Pill decreased significantly?P<0.01?.The level of serum estradiol in tamo-xifen group decreased significantly?P<0.01?.The level of progestogen in tamoxifen group,the high and middle dose groups of Xihuang Pill increased significantly?P<0.05 or P<0.01?.The index of uterine organs in tamoxifen group,the high and middle dose groups of Xihuang Pill decreased significantly?P<0.05 or P<0.01?.The index of ovarian organs in tamoxifen group and the high dose group of Xihuang Pill decreased significantly?P<0.01?.2.From the perspective of pituitary tissue,compared with the blank group,the le-vels of follicle stimulating hormone,follicle stimulating hormone gene expression and gonadotropin-releasing hormone receptor protein expression in pituitary tissue in tam-oxifen group and Xihuang pill groups decreased significantly?P<0.01?.The levels of luteinizing hormone in tamoxifen group and Xihuang pill groups increased significan-tly?P<0.05 or P<0.01?.The expression level of gonadotropin-releasing hormone recep-tor gene in the middle dose group of Xihuang Pill decreased significantly?P<0.05?.The expression level of luteinizing hormone gene in tamoxifen group and the high dose group of Xihuang Pill increased significantly?P<0.01?.The ultrastructure is shown in the text.3.From the ovarian tissue,compared with the blank group,the level of estradiol in ovarian tissue in tamoxifen group,Xihuang pill high and middle dose groups decr-eased significantly?P<0.01?.The levels of progestogen and luteinizing hormone rece-ptor gene expression in tamoxifen group and Xihuang pill groups increased signifi-cantly?P<0.01?.The expression level of follicle stimulating hormone receptor gene in tamoxifen group and Xihuang pill groups decreased significantly?P<0.01?.The ex-pression level of follicle stimulating hormone receptor protein in tamoxifen group and the high and middle dose groups of Xihuang pill decreased significantly?P<0.01?.The expression level of luteinizing hormone receptor protein in the high,middle and low dose groups of Xihuang pill increased significantly?P<0.01?.Compared with the blank group,the expression of estradiol receptor?and estradiol receptor?in ovarian tissues in tamoxifen group and Xihuang pill high and middle dose groups decreased significantly?P<0.05 or P<0.01?.The expression of progestogen receptor in ovarian tissues in the high and middle dose groups of Xihuang pill increased significantly?P<0.01?.The ovarian tissues HE staining and ultrastructures of under transmission electron microscopy are shown in the text.Conclusion:1.Xihuang pill reduces the index of uterus and ovary organs,reduces the content of gonadotropin releasing hormone,follicle stimulating hormone and estradiol in rat serum,and increases the content of luteinizing hormone and progesterone,which indi-cates that Xihuang pill affects the level of hypothalamus-pituitary-ovary axis related hormone in blood and inhibits the function of uterus and ovary.2.Xihuang pill can cause pituitary ultrastructure disorder,reduce follicle stimul-ating hormone content in rat pituitary,increase luteinizing hormone content,inhibit follicle stimulating hormone gene expression,gonadotropin releasing hormone rece-ptor gene and protein expression,and enhance luteinizing hormone gene expression,which indicates that Xihuang pill has intervention effect on pituitary function and ultrastructure.3.Xihuang pill can cause ovarian ultrastructure disorder,inhibit expression of follicle stimulating hormone receptor gene and protein,promote expression of lute-inizing hormone receptor gene and protein,reduce estradiol content in rat ovary,increase progesterone content,inhibit expression of estradiol receptor?and?,and promote expression of progesterone receptor,which indicates that Xihuang pill has intervention effect on ovarian function and causes ovarian tissue atrophy.