| Background As the ovarian aging, women not only lost their reproduction character, but also suffered an perimenopausal period syndrome include hot fish, atrophy of genitourinary tract, subsidence of the nerves system, risk of cardiovascular diseases and fractures, hazarding the physical and mental health of the women. In our previous study, several relative genes relating to the ovarian aging process had been found by a gene chips technique. Now we certified the expression of the key genes in ovarian aging process in both mRNA and protein levels, and explored the possible function and mechanism of the relative genes in ovarian aging process by a series study of in vivo and in vitro.MethodReal-time polymerase chain reaction and western blot were used to detect the mRNA and protein expression of the key genes in different stages of age and gonadotropin-primed inmature C57BL/6j mouse ovaries. Then we decreased the mRNA expression of the key gene in ovary by intrabursal injection of the siRNA to the key genes in vivo, and regulated the downstream cell signal pathway of the key genes in vitro by the using of the monoclonal antibody and recombination protein to the key genes in vitro. H-E stain, terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL), western blot, enzyme linked fluoreseent assay(ELFA), real-time Polymerase Chain Reaction, immunofluorescence were used to detect the apoptosis, hormone secretion and relative mRNA, protein expression of the granulose cell. The light microscope and confocal laser scanning microscope were used for the detecting of the follicle stage inform and apoptosis, functional protein expressing in the ovaries. Result(1) The mRNA and protein decreased in the aging process of the mouse ovary. In the gonadotropin-primed inmature C57BL/6j mouse ovaries, the expression of the898a mRNA and protein decreased significantly4hours after the serum gonadotrophin injection, but increased step by step in the48hours for injection. Then as the human chorionic gonadotrophin injection, the expression of898a diminished strictly at4hours but stepped up soon at the time for ovulation.(2) As decreased the mRNA expression of898a in vivo by intrabursal injection of siRNA, the quantity of the atresia follicle and apoptosis granulose cell increased significantly and the primordial follicle decreased notably. In addition, the functional gene of the oocyte such as the GDF9, BMP15, connexin37,ZP3were suppressed and the proliferation and differentiation of the granulose cells were promoted significantly as the mRNA of3HSD, CYP11a, CYP19a, STAR, connexin43, PCNA increased notably. And the expression of the AMH decreased significantly as well.(3) The898a depressed the hormone secretion and relative mRNA expression of the steroid hormone synzymes especially the progesterone, as the AMH were increased notably in vitro.(4) The expression of Peroxiredoxins diminished in the mouse ovarian aging process. In the gonadotropin-primed inmature C57BL/6j mouse ovaries, the PRDX â… mRNA stepped up as the PMSG injection, but decreased as the HCG injection till the ovulation and nonage of corpus luteum. Then there were no difference in the ovaries of the PRDX â… mRNA expression in the oestrous cycle. In the gonadotropin-primed inmature C57BL/6j mouse ovaries, the PRDXâ…¢ mRNA diminished step by step in the inmature ovaries as the PMSG and HCG injection till the ovulation to the lowest, but increased after ovulation and the nonage of the corpus luteum. Then there were no difference in the ovaries of the PRDX â…¢ mRNA expression in the oestrous cycle. In the gonadotropin-primed inmature C57BL/6j mouse ovaries, the PRDXâ…£ mRNA increased as the PMSG injection but decreased as the ovulation and the nonage of the corpus luteum. Then there were no difference in the ovaries of the PRDXIV mRNA expression in the oestrous cycle. In the gonadotropin-primed inmature C57BL/6j mouse ovaries, the PRDXV mRNA increased significantly as the PMSG injection and the nonage of the corpus luteum, but decreased before the ovulation. Then the expression of the PRDXâ…¤ mRNA was higher in the estrus and metestus mouse ovaries than the diestrus and proestrus. In the gonadotropin-primed inmature C57BL/6j mouse ovaries, the PRDXâ…¥ mRNA increased as the PMSG injection and the nonage of the corpus luteum, but diminished around the ovulation. Then the expression of the PRDXâ…¥ mRNA was higher in the proestrus mouse ovaries than the diestrus and metestus.(5) The expression of NRP-1mRNA decreased as the mouse ovarian aging, and was much higher in the newborn mouse ovaries as the period of primordial follicle formation than the period of primary and secondary follicle formation. In the gonadotropin-primed inmature C57BL/6j mouse ovaries, the expression of NRP-1mRNA decreased as the PMSG injection but stepped up at the48hours. Then the expression continued step up as the HCG injection till the ovulation and decreased as nonage of the corpus luteum.Conclusion(1) The898a was the relative gene of the ovarian aging process and acted importantly in the follicle development process.(2) As the deficiency of898a, the atresia follicle increased as the excessus differentiation of the granulose cell and suppression of the oocyte functional, and the primordial follicle decreased as the diminished of the AMH levels.(3) The898a support the secretion of AMH by suppressing the terminal differentiation of the granulose cell.(4) The PRDXs play important roles in ovarian aging and follicle development.(5) The NRP-1may regulate the ovarian aging process through the initial recruitment and cycle recruitment. |
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