卵巢低反应女性中沉默信息调节因子1对卵巢颗粒细胞的调控

doi:10.3969/j.issn.1006-5725.2023.21.004

Abstract

Abstract:

Objective To investigate the regulation of SIRT1 on ovarian granulosa cell in women with poor ovarian response. Methods A total of 60 women who underwent IVF/ICSI-ET were included in this study， 30 women in poor ovarian response （POR） group and 30 women in normal ovarian response （NOR） group. The granulosa cells in follicular fluid were isolated and purified by density gradient centrifugation. The granulosa cells of POR group were treated with SIRT1 agonist resveratrol （RESV） or inhibitor EX527， respectively， and the control group was granulosa cells treated without drugs. mRNA levels of SIRT1 and key enzymes of steroid hormone synthesis （StAR， CYP19， CYP17） in granulosa cells were detected by qPCR. The levels of SIRT1， steroid hormone synthesis key enzymes protein and apoptosis-related protein （Bcl-2， Caspase-3） in granulosa cells were detected by Western blot. The apoptosis rate of granulosa cells was detected by TUNEL method. Determination of estradiol in granulosa cell culture medium by chemiluminescence. Results The mRNA and protein levels of SIRT1 in granulosa cells in POR group were lower than those in NOR group （P = 0.003， P < 0.001）. RESV up-regulated SIRT1 mRNA and protein levels （P < 0.001）， up-regulated mRNA and protein levels of steroid hormone synthesis key enzyme （P < 0.05）. EX527 down-regulated SIRT1 mRNA and protein levels （P = 0.023， 0.001）， down-regulated mRNA and protein levels of steroid hormone synthesis key enzymes （P < 0.05）. After RESV treatment， the apoptosis rate of granulosa cells was decreased （P < 0.001）， the Bcl-2 protein level was increased （P < 0.001）， and the Caspase-3 protein level was decreased （P < 0.001）. After EX527 treatment， the apoptosis rate of granulosa cells was increased （P < 0.001）， the Bcl-2 protein level was decreased （P = 0.003）， and the Caspase-3 protein level was increased （P < 0.001）. Conclusion The SIRT1 level in granulosa cells of POR women was lower than that of NOR women. RESV up-regulated SIRT1 expression in granulosa cells. SIRT1 inhibited granulosa cell apoptosis， increased the level of steroid hormone synthesis key enzyme， and promoted estradiol synthesis in granulosa cell. Therefore， SIRT1 activators such as RESV may be a therapeutic agent to improve ovarian reactivity and increase the number of oocytes obtained in women with POR.

Key words: poor ovarian response, silent information regulator 1, ovarian granulosa cell, apoptosis

CLC Number:

R711.75

Hui KE,Lizheng YAN,Yu ZHANG,Jilong MAO,Yufei YAO. Regulation of ovarian granulosa cell by SIRT1 in women with poor ovarian response[J]. The Journal of Practical Medicine, 2023, 39(21): 2709-2717.

Figures/Tables 17

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项目	POR组	NOR组	t值	P值
年龄（岁）	32.26 ± 1.87	32.26 ± 1.87	-	-
体质量指数（kg/m²）	20.66 ± 0.89	20.36 ± 1.11	1.143	0.258
HCG日E₂水平（pmol/L）	2 370 ± 539	11 502 ± 2 527	-19.352	< 0.001
获卵数	2.73 ± 0.45	10.33 ± 1.88	-21.517	<0.001

基因	引物序列
SIRT1
正向Forward	5'-GGAGCAGATTAGTAGGCGGC-3'
反向Reverse	5'-CTTGGACTCTGGCATGTCCC-3'
CYP17
正向Forward	5'-TGGCTCTCTTGCTGCTTACC-3'
反向Reverse	5'-GGTGGCCGACAATCACTGTA-3'
CYP19
正向Forward	5'-TGTGCAATGTGCTGTGGATTC-3'
反向Reverse	5'-TCTCAATGAAGGCATGGGGG-3'
StAR
正向Forward	5'-CAGACTTCGGGAACATGCCT-3'
反向Reverse	5'-GGGACAGGACCTGGTTGATG-3'
β?actin
正向Forward	5'-TTGTTACAGGAAGTCCCTTGCC-3'
反向Reverse	5'-ATGCTATCACCTCCCCTGTGTG-3'

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Regulation of ovarian granulosa cell by SIRT1 in women with poor ovarian response

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