摘要
目的探讨光污染大鼠、正常光照大鼠与滋阴补阳方序贯法干预后的大鼠卵巢的microRNA测序表达谱的差异情况。方法将80只SD雌性大鼠随机分为对照组25只和造模组55只,对照组大鼠25只给予12/12h光/暗循环的光照条件,造模组大鼠55只给予24 h持续光照的条件,造模时间定为50天,剔除造模失败者5只,剩余造模大鼠随机分为模型组25只和实验组25只。对照组、模型组予以生理盐水灌胃,实验组予以滋阴补阳方序贯法。取三组大鼠卵巢,采用Illumina HiSeq 2500高通量测序技术进行测序,比较三组的microRNA表达差异。对差异表达的microRNA进行筛选,并进行GO和KEGG功能显著性富集分析。荧光定量RT-PCR对差异表达microRNA进行验证。结果本研究发现光污染组和正常光照组组间呈显著性差异表达的16个已知的microRNA,其中5个microRNA表达上调,11个microRNA表达下调。光污染组和实验组组间呈显著性差异表达的4个已知的microRNA,其中3个microRNA表达上调,1个microRNA表达下调。GO分析显示差异表达的microRNA在富集的分子功能上多集中于生物调节、生长过程、分子的结合等方面,KEGG通路富集分析示,受microRNA调控的靶基因参与的这些通路中多集中于神经组织受体相互作用、癌症、雌激素、MAPK等相关信号通路。荧光定量RT-PCR结果示差异性miRNA的表达趋势与测序结果相一致。结论差异表达的microRNA可能在光污染环境下对卵巢的生长发育具有重要的作用,滋阴补阳方序贯法可能通过这些相关信号通路作用于卵巢组织起到改善卵巢功能的作用。
Objective To explore the difference of the expression profiles of ovarian microRNA sequencing of lightpollution rats, normal light rats, light pollution rats after the nourishing Yin and tonifying Yang sequential traditionalChinese medicine treatment. Methods The 80 female rats were randomly divided into two groups: 25 rats in the controlgroup and 55 rats in the modeling group. 25 rats in the control group were exposed to 12/12 hour light/dark cycles, while55 rats in the modeling group were continuously exposed to 24-hour light. The selected light intensity was 300 ± 20 lux,and modeling time was 50 days. 5 rats did not meet the criterion and were excluded from the experiment, while theremaining 50 rats were assigned as 25 rats of the model group and 25 rats of the case group. The normal group and themodel group were given the saline lavage, while the case group was given the nourishing Yin and tonifying Yangsequential traditional Chinese medicine lavage. Three groups of rat ovaries were taken, and Illumina HiSeq 2500 high-throughput sequencing technology was used to sequence the three groups of microRNA expression differences. Thedifference expression of microRNA was screened and the target gene was predicted. And the function of GO and KEGGwas analyzed. Fluorescence quantitative RT-PCR was used to verify the differential expression of microRNA.Results The expression of 16 known microRNAs between light pollution rats and normal light rats was significantlydifferent. Among these 16 microRNAs, 5 microRNAs were upregulated and 11 microRNAs were downregulated. Theexpression of 4 known microRNAs between light pollution rats and traditional Chinese medicine rats was significantlydifferent. Among these 4 microRNAs, 3 microRNAs were upregulated and 1 microRNA was downregulated. The GOanalysis shows that the samples are mainly enriched in biological regulation, the process of growth, and molecularbinding in terms of enriched molecular functions. The KEGG Pathway Enrichment Analysis shows that pathways thatinvolve microRNA regulated target genes are mainly enriched in the interaction of nerve tissue receptors, cancer,estrogen and MAPK related signaling pathways. Fluorescence quantitative RT-PCR results revealed that the trend of theexpression change of four microRNAs was consistent with the results of high throughput sequencing. Conclusion Thedifferential expression of microRNAs may play an important role in the growth and development of ovary in the lightpollution environment. The nourishing Yin and tonifying Yang sequential traditional Chinese medicine may improveovarian function by these signal pathways.
作者
马倩雯
谈勇
Ma Qianwen;Tan Yong(Zhenjiang Affiliated Hospital of Nanjing University of Chinese Medicine,Zhenjiang hospital of TCM,Gynecology,Zhenjiang,China,212000;Nanjing University of Chinese Medicine First clinical medical college,Nanjing,China,210000)
出处
《世界科学技术-中医药现代化》
CSCD
北大核心
2019年第11期2499-2507,共9页
Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology
基金
国家自然科学基金委员会面上项目(81373683):人工光环境对卵巢发育和节律的影响及中药干预作用的研究,负责人:谈勇
镇江市科技局镇江市卫生计生科技重点专项项目重点专项项目(SHW2017016):滋阴补阳方序贯法联合氯米芬治疗日节律紊乱性排卵障碍的临床研究,负责人:马倩雯。
关键词
光污染
卵巢
MICRORNA
测序
滋阴补阳方序贯法
light pollution
ovary
microRNA
sequencing
nourishing Yin and tonifying Yang sequential traditional Chinese medicine
