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中国临床药理学与治疗学 ›› 2019, Vol. 24 ›› Issue (10): 1101-1106.doi: 10.12092/j.issn.1009-2501.2019.10.004

• 基础研究 • 上一篇    下一篇

B3型柯萨奇病毒感染对胰岛细胞糖尿病相关LncRNA表达的影响

沈雅婧1,李兰娟2     

  1. 1浙江大学医学院附属第一医院传染病诊治国家重点实验室, 2感染性疾病诊治协同创新中心,杭州 310058,浙江
  • 收稿日期:2019-04-15 修回日期:2019-05-08 出版日期:2019-10-26 发布日期:2019-10-28
  • 通讯作者: 李兰娟,女,博士,教授,博士生导师,研究方向:传染病。
  • 作者简介:沈雅婧,女,本科,主治医师,研究方向:全科医学,传染病。 Tel:15988833621 E-mail:408000394@qq.com
  • 基金资助:

    国家自然科学基金重大项目(81790631)

Effects of Coxsackievirus B3 infection on the expression of diabetes-related LncRNA in islet cells

SHEN Yajing1, LI Lanjuan2   

  1. 1 State Key Laboratory of Diagnosis and Treatment of Infectious Diseases, 2 Infectious Diseases Diagnosis and Treatment Collaborative Innovation Center, the First Affiliated Hospital of Zhejiang University, Hangzhou 310058, Zhejiang, China
  • Received:2019-04-15 Revised:2019-05-08 Online:2019-10-26 Published:2019-10-28

摘要:

目的:探讨B3型柯萨奇病毒诱发糖尿病发生的具体机制。方法:采用B3型柯萨奇病毒感染人胰岛细胞HUM-CELL-0058,病毒感染48 h后提取细胞总RNA并通过荧光定量PCR检测糖尿病相关LncRNA。对调控三种LncRNA转录的转录因子mRNA及蛋白采用实时定量PCR及Western blot进行监测。构建了PAX6及NF-κB的过表达质粒,并转染病毒感染后的胰岛β细胞,检测柯萨奇病毒感染后异常表达LncRNA的表达水平。结果:糖尿病相关LncRNA,Lnc-P12792和MALAT1有上调表达,上升2~3倍(P<0.01),而HI-Lnc45的表达水平约为对照组的60%(P<0.01)。实时定量PCR及Western blot结果表明调控HI-Lnc45表达的转录因子PAX6表达量降低50%而调控Lnc-P12792和MALAT1的转录因子NF-κB(p65)表达升高2.5倍(P<0.001),这与其调控的转录因子趋势一致。表明LncRNA的表达差异是由于该基因转录因子表达异常所导致。实时定量结果表明,三种异常表达LncRNA的mRNA水平回归B3型柯萨奇病毒感染前的正常水平,证明这一趋势可被调控这两种转录因子的表达量所拯救。结论:B3型柯萨奇病毒感染人胰岛β细胞会引起转录因子NF-κB的上调表达以及PAX6的下调表达,并引起下游与糖尿病发生有关LncRNA的异常表达,这可能是B3型柯萨奇病毒诱发糖尿病发生的病理机制之一。

关键词: B3型柯萨奇病毒, 糖尿病, LncRNA, 转录因子

Abstract:

AIM: To explore the specific mechanism of diabetes mellitus induced by Coxsackievirus B3.  METHODS: Human islet cells HUM-CELL-0058 were infected with Coxsackievirus B3. Total RNA was extracted 48 hours after infection and LncRNA related to diabetes mellitus was detected by fluorescence quantitative PCR. Real-time quantitative PCR and Western blot were used to monitor the transcription factors and proteins regulating the transcription of three LncRNA. The over-expression plasmids of PAX6 and NF-kappa B were constructed and transfected into B cells infected with the virus to detect the abnormal expression of LncRNA after coxsackievirus infection. RESULTS:Diabetes-related LncRNA, Lnc-P12792 and MALT1 were up-regulated, while HI-Lnc45 was down-regulated. Real-time quantitative PCR and Western blot results showed that the expression of PAX6, a transcription factor regulating HI-Lnc45 expression, decreased while the expression of NF-kappa B, a transcription factor regulating Lnc-P12792 and MALT1, increased, which was consistent with the trend of transcription factors regulated by PAX6, indicating that the expression difference of LncRNA was due to the abnormal expression of transcription factors of the gene. Real-time quantitative results showed that the three abnormal expression levels of LncRNA returned to the normal level before coxsackievirus B3 infection, which proved that this trend could be saved by regulating the expression of these two transcription factors. CONCLUSION: Coxsackievirus B3 infection of human pancreatic islet β cells can induce up-regulation of transcription factor NF-kappa B and down-regulation of PAX6, and induce abnormal expression of LncRNA related to diabetes mellitus downstream, which may be one of the pathological mechanisms of Coxsackievirus B3 induced diabetes mellitus.

Key words: type B3 Coxsackie virus, diabetes, LncRNA, transcription factor

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