H12、RS13和RL6在miR-199b-5p敲除小鼠卵巢中的表达研究

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摘要目的探讨组蛋白H1变体(H12)、核糖体蛋白S13(RS13)和核糖体蛋白L6(RL6)在miR-199b-5p敲除小鼠卵巢中的表达变化。方法 (1)自行繁育miR-199b-5p敲除(Knockout,KO)和野生型(Wild-type,WT)C57BL/6小鼠,分别记录两组雌鼠产仔情况(2)基因型鉴定验证分组,取性成熟期小鼠(鼠周龄为11~13周)卵巢用于验证实验。(3)用实时荧光定量PCR(qPCR)检测每组小鼠(n=10)卵巢组织中H12、RS13和RL6的mRNA表达水平变化。(4)用蛋白免疫印迹法(WB)检测每组小鼠(n=3)卵巢组织中H12、RS13和RL6的蛋白表达水平变化。结果 (1)共统计半年内两组雌鼠产仔情况:WT组生产27窝,平均每窝产仔数为(8.15±0.41)只;KO组生产28窝,平均每窝产仔数为6.43±0.32只,与WT组比较,KO组雌鼠产仔数显著下降(P=0.004),差异有统计学意义。(2)qPCR结果示:与WT组比较,KO组卵巢组织中H12(P=0.038)、RS13(P=0.011)和RL6(P=0.009)的mRNA表达水平显著升高,差异均有统计学意义。(3)WB结果示:与WT组比较,KO组小鼠卵巢组织中H12(P= 0.014)蛋白表达水平显著升高,而 RS13(P= 0.005)和RL6(P=0.009)蛋白表达水平显著下降,差异均有统计学意义。结论敲除miR-199b-5p可能影响H12、RL6和RS13基因的表达使小鼠卵巢储备能力下降。

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	袁秀秀
	王煜

关键词 ： miR-199b-5p, 基因敲除, 卵巢, 小鼠

Abstract：Objective To investigate the influence of miR-199b-5p knockout on ovarian reproductive capacity in mice, and to verify the expression changes of candidate genes which are histone H1 variants(H12), ribosomal protein S13(RS13) and ribosomal proteins L6(RL6) in the ovaries of miR-199b-5p knockout mouse.Methods The miR-199b-5p knockout(KO) and Wild-type(WT) C57BL/6 mice were self-bred, and the litter size of female mice was recorded respectively.(2)Genotype identification to verify the group, the ovaries of sexually mature mice(11-13 weeks old were taken for verification experiments.(3)The mRNA expression levels of H12, RS13 and RL6 genes in each group mouse of ovarian tissues(n=10)were detected by real-time quantitative PCR(qPCR). The protein expression levels of H12, RS13 and RL6 in each group mouse of ovarian tissues(n=3) were detected by western blot(WB).Results (1)We observed 27 litters of WT group and 28 litters of KO group in six months, respectively its average litter size were (8.15±0.41) and (6.43 ±0.32). Interestingly, the litter size of KO group females was significantly reduced(P=0.004).(2) Compared with WT group, the mRNA expression levels of H12(P=0.038), RS13(P=0.011) and RL6(P=0.009) were significantly increased by qPCR.(3)The protein expression level of H12(P=0.014) was visibly increased, while RS13(P= 0.005) and RL6(P=0.009) were distinctly declined by WB.Conclusions The study demonstrates miR-199b-5p knockout may affect the expression of H12, RL6 and RS13 to induce the decrease of the ovarian reproductive capacity in mice.

Key words： miR-199b-5p knockout of gene ovarian mouse

收稿日期: 2022-03-17

ZTFLH:

R71

基金资助:内蒙古自治区草原英才创新团队项目(2022TD014)、内蒙古科技计划项目(2023KJHZ0038)、内蒙古医科大学致远人才计划(ZY020238)、内蒙古医科大学科技成果转化项目(YKD2020CGZH007)、内蒙古医科大学后续科研项目(2020)、内蒙古自治区高等学校“青年科技英才”支持计划(NJYT-18-B12)、白求恩-默克糖尿病研究基金(2017)

通讯作者: 王煜(wuai1544@163.com)

引用本文:

袁秀秀, 王煜. H12、RS13和RL6在miR-199b-5p敲除小鼠卵巢中的表达研究[J]. 中国生育健康杂志, 2024, 35(1): 48-53. YUAN Xiuxiu, WANG Yu. Expression of H12, RS13 and RL6 in ovarian tissues of miR-199b-5p knockout mouse. Chinese Journal of Reproductive Health, 2024, 35(1): 48-53.

链接本文:

http://cjrh.bjmu.edu.cn/CN/ 或 http://cjrh.bjmu.edu.cn/CN/Y2024/V35/I1/48

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