新温肾生精饮对精子和早期胚胎中H3K9me2修饰水平的影响  

Effect of New Wenshen Shengjing Decoction on the modification levels of H3K9me2 in the sperms and early embryos

作　　者：陈强根 孙艳美[1] 葛莹 张金宇 韩毅 吴依婷 王雪楠[3] 潘晓燕[1] CHEN Qiang-gen;SUN Yan-mei;GE Ying;ZHANG Jin-yu;HAN Yi;WU Yi-ting;WANG Xue-nan;PAN Xiao-yan(Jilin Medical University,Jilin Jilin,132013,China;Jilin Gynecology and Obstetrics Hospital,Jilin Jilin 132011,China;Affiliated Hospital of Jining Medical College,Jining Shandong 272029,China)

机构地区：[1]吉林医药学院,吉林吉林132013 [2]吉林市妇产医院,吉林吉林132011 [3]济宁医学院附属医院,山东济宁272029

出　　处：《时珍国医国药》2022年第1期79-83,共5页Lishizhen Medicine and Materia Medica Research

基　　金：吉林省科技发展计划项目(20190304071YY);吉林省卫生与健康青年科技骨干培养计划(2018Q047);吉林市科技创新发展计划项目(201831721、20190403224);吉林省教育厅“十三五”科学技术研究项目[JJKH20191065KJ、JJKH20200454KJ];山东省中医药科技发展计划项目(2019-0463);吉林省大学生创新创业训练计划项目(201913706031、202013706019)。

摘　　要：目的探讨新温肾生精饮(New Wenshen Shengjing Decoction,NWSSJD)对精子和早期胚胎中H3K9me2修饰水平的影响。方法将性成熟的雄性小鼠环磷酰胺(cyclophosphamide,CPA)造模后,灌服30天的NWSSJD,然后与超排处理的母鼠交配,统计各期胚胎发育率;免疫荧光染色法检测精子和各期胚胎中H3K9me2的表达;qRT-PCR分析囊胚细胞中凋亡相关基因BCL-2和P53的表达。结果与CPA组相比,NWSSJD显著降低了精子中H3K9me2的表达水平,显著提高了早期胚胎(2-细胞胚胎、3-4细胞胚胎、8-16细胞胚胎和囊胚)的发育率,显著促进了BCL-2基因表达,而降低了P53基因表达,抑制了囊胚细胞凋亡。结论NWSSJD通过维持精子和早期胚胎中正常H3K9me2修饰水平,抑制胚胎细胞凋亡,而促进早期胚胎发育。Objective New Wenshen Shengjing Decoction(NWSSJD)is a traditional compound Chinese medicine,mainly including Panax ginseng,Cornu Cervi Nippon Parvum,Cynomorium songaricum,Radix Astragali,Epimedium brevicornum and Angelica sinensis,which has an significant therapeutic effect on male spermatogenesis disorder.Some components in NWSSJD can change epigenetic modifications in the development of different types of cells,but it is unknown whether it will cause epigenetic changes in the sperms or early embryos.Methods Mature male mice were gavaged with NWSSJD for 30 days,following the treatment with cyclophosphamid(CPA),and then mated with the superovulated female mice.The harvested embryo development rate of each stage was calculated.The expressions of H3 K9 me2 in the sperms and early embryos were examined by immunofluorescence staining,and the expressions of apoptosis-related gene BCL-2 and P53 in the blastocysts were analyzed by qRT-PCR.Results Compared with CPA group,NWSSJD significantly decreased the expression of H3 K9 me2 in the sperms,and significantly increased the development rates of early embryos(2-cell embryos,3-4-cell embryos,8-16-cell embryos and blastocysts).Moreover,the expression of BCL-2 gene was greatly increased,while the expression of P53 gene was significantly lowered,leading the inhibition of the apoptosis of blastocysts in NWSSJD group.Conclusion These results suggest that NWSSJD can promote the development of early embryos by maintaining the normal levels of H3 K9 me2 in the sperms and early embryos and inhibiting apoptosis of embryonic cells.

关 键 词：新温肾生精饮 精子 早期胚胎 H3K9me2 环磷酰胺 

分 类 号：R285.5[医药卫生—中药学]