| 张蓉,张晓艳.基于二代测序用于瘦素缺陷T2DM ob/ob小鼠与C57BL/6小鼠的转录组差异分析.[J].中南医学科学杂志.,2021,(5):512-517. |
| 基于二代测序用于瘦素缺陷T2DM ob/ob小鼠与C57BL/6小鼠的转录组差异分析 |
| Second generation sequencing was used to analyze transcriptome differences between LEPtin-deficient T2DM ob/ob mice and C57BL/6 mice |
| 投稿时间：2020-12-21 修订日期：2021-01-30 |
| DOI：10.15972/j.cnki.43-1509/r.2021.05.004 |
| 中文关键词: 二代测序 瘦素缺陷 2型糖尿病 C57BL 差异性基因 |
| 英文关键词:second generation sequencing LEPtin-deficient T2DM C57BL difference gene |
| 基金项目:作者简介:张蓉,硕士研究生,主管技师,研究方向为蛋白质组学和分子生物学,E-mail为51608293@qq.com。通信作者张晓艳,博士,研究方向为病理学与病理生理学,E-mail为zhangxy1120@aliyan.com。 |
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| 中文摘要: |
| 目的探讨瘦素缺陷T2DM ob/ob小鼠与C57BL/6小鼠胰腺组织差异性基因及相关的生物学功能。方法取8周雌雄性ob/ob小鼠和C57BL/6小鼠,饲养2周后取胰腺组织进行转录组测序。采用二代测序进行转录组测序。使用edgeR包分析两小鼠胰腺组织的差异基因,使用DAVID数据库进行差异基因生物学功能及信号通路分析；寻找hub基因,分析基因间相关性及两组表达量差异。结果取瘦素缺陷T2DM ob/ob小鼠和C57BL/6J小鼠的胰腺组织进行病理切片HE染色,观察ob/ob小鼠较正常小鼠胰岛增生、肥大,岛内β细胞核增多、密集,胞质减少,排列紊乱；胰腺组织提取的RNA进行转录组测序,经过滤、质控、比对后,ob/ob较C57BL/6J小鼠上调基因88个,下调基因94个,共182个；差异基因生物学功能和信号通路主要涉及翻译、rRNA加工、核糖体、细胞内核糖核蛋白复合物、胞质小核糖体亚基、粘着斑、细胞外泌体、核糖体的结构成分、RNA结合、mRNA结合和核糖体信号通路等；寻找hub基因,RPS12-ps3、Rps4x、Gm7536、Rpl7、Rpl38、Rps7、Rps15a、Rps3a1、Rpl17和Rps27a,除Rpl7与Rps27a呈负相关外,其余均成正相关,hub基因在ob/ob小鼠中均较C57BL/6J小鼠低表达,差异具有显著性(P＜0.05)。结论ob/ob小鼠和C57BL/6J小鼠差异基因主要是核糖体蛋白基因,核糖体蛋白基因水平的降低可能影响rRNA加工中的关键蛋白的功能,从而可能导致T2DM的发生。 |
| 英文摘要: |
| To investigate the difference gene and related biological function of pancreatic tissue between LEPtin deficient T2DM ob/ob mice and C57BL/6 mice. MethodsEight-week-old male and female ob/ob mice and C57BL/6 mice were taken. After feeding for 2 weeks, pancreas tissues were taken for transcriptome sequencing. Second generation sequencing was used for transcriptome sequencing. EdgeRpackage was used to analyze the differentially expressed genes in the pancreatic tissues of the two mice. DAVID database was used to analyze the biological function and signal pathway of the differentially expressed genes. Hub genes were found to analyze the correlation between genes and the expression difference between the two groups. ResultsHE staining was performed on the pancreatic tissues of leptin deficient T2DM ob/ob mice and C57BL/6J mice, and pancreatic islet hyperplasia and hypertrophy were observed in ob/ob mice compared with normal mice. The nuclei in the islands were increased and dense, with decreased cytoplasm and disordered arrangement. Transcriptomic sequencing was performed on the RNA extracted from pancreatic tissues. After filtration, quality control and comparison, 88 up-regulated genes and 94 down-regulated genes were found in ob/ob compared with C57BL/6J mice, a total of 182 genes. Subsequently, the biological functions and signaling pathways of different genes mainly involve translation, rRNA processing, ribosomes, intracellular ribosomal protein complexes, cytoplasmic small ribosome subunits, adhesion spots, exosomes, structural components of ribosomes, RNA binding, mRNA binding and ribosome signaling pathways, etc. Hub genes were found, rPS12-PS3, Rps4x, Gm7536, Rpl7, Rpl38, Rps7, Rps15a, Rps3a1, Rpl17 and Rps27a were all positively correlated, except that Rpl7 was negatively correlated with Rps27a. The expression of hub genes in ob/ob mice was lower than that in C57BL/6J mice, and the difference was statistically significant (P＜0.05). ConclusionThe difference genes between ob/ob mice and C57BL/6J mice are mainly ribosomal protein genes, and the reduction of ribosomal protein gene level may affect the function of key proteins in rRNA processing, which may lead to the occurrence of T2DM. |
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