上海交通大学学报（医学版）

上海交通大学学报（医学版）

• 论著（基础研究） • 上一篇    下一篇

抑制骨髓间充质干细胞内源性白介素-6分泌对氧糖剥夺损伤PC12细胞的影响

何慕兰，刘晶晶，谷 燕，李廷玉，陈 洁   

  1. 重庆医科大学附属儿童医院儿童营养研究中心， 儿童发育疾病研究教育部重点实验室， 重庆市干细胞治疗工程技术研究中心， 重庆 400014
  • 出版日期:2014-10-28 发布日期:2014-10-28
  • 通讯作者: 陈 洁, 电子信箱: jchen010@cqmu.edu.cn。
  • 作者简介:何慕兰（1987—）， 女， 壮族， 硕士生; 电子信箱: 896717643@qq.com。
  • 基金资助:

    国家自然科学基金（81271385）；重庆市卫生局重点项目（渝2009-1-41）

Effects of inhibiting secretion of mesenchymal stem cells originated interleukin-6 on oxygen glucose deprivation injured PC12 cells

HE Mu-lan, LIU Jing-jing, GU Yan, LI Ting-yu, CHEN Jie   

  1. Children's Nutrition Research Center, Key Laboratory of Ministry of Education in Childhood Development Diseases, Children's Hospital of Chongqing Medical University, Chongqing Stem Cell Therapy Engineering Technical Center, Chongqing 400014, China
  • Online:2014-10-28 Published:2014-10-28
  • Supported by:

    National Natural Science Foundation of China, 81271385; Key Project of Chongqing Health Bureau, 2009-1-41

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摘要：

目的 探讨骨髓间充质干细胞(MSCs)来源的白介素-6 （IL-6）对神经细胞损伤的作用。方法 构建大鼠沉默IL-6 （siIL-6）慢病毒，感染MSCs，嘌呤霉素筛选siIL-6-MSCs稳定细胞株。Real-Time PCR和Western blotting检测细胞IL-6 mRNA和蛋白表达变化，ELISA检测siIL-6-MSCs培养上清液中IL-6分泌水平。将siIL-6-MSCs细胞与氧糖剥夺（OGD）损伤的PC12细胞分离共培养，Annexin V-FITC/PI法检测损伤细胞凋亡，Western blotting检测细胞中IL-6信号通路关键蛋白STAT3以及凋亡相关因子Bax和Bcl-2蛋白的表达。结果 siIL-6干扰序列正确插入慢病毒质粒中。siIL-6慢病毒感染MSCs后，IL-6 mRNA和蛋白表达及IL-6分泌水平均显著降低。与siIL-6-MSCs分离共培养后，OGD损伤的PC12细胞凋亡增加，STAT3、Bcl-2蛋白表达显著降低，Bax蛋白表达显著升高。结论 MSCs来源的IL-6可能通过STAT3信号通路，调节OGD损伤PC12细胞的抗凋亡作用。

关键词: 骨髓间充质干细胞, 白介素-6, 氧糖剥夺, 神经损伤, STAT3, 抗凋亡

Abstract:

Objective To investigate the effects of mesenchymal stem cells (MSCs) originated interleukin-6 (IL-6) on the nerve cells injury. Methods The rat siIL-6 recombinant lentivirus plasmid was constructed and the siIL-6 recombinant lentivirus was infected by MSCs. The MSCs cell line with stable siIL-6 expression was selected by puromycine. The variations of IL-6 mRNA and protein expression levels were detected by the Real-Time PCR and Western blotting. The secretion level of IL-6 was detected from the cultured supernatant of siIL-6-MSCs by the ELISA. The siIL-6-MSCs cells and PC12 cells injured by oxygen glucose deprivation (OGD) were separated and co-cultured. Cell apoptosis was detected by the Annexin V-FITC/PI. Expressions of key protein STAT3 of IL-6 signaling pathway and apoptosis related factors Bax and Bcl-2 were detected by the Western blotting. Results The IL-6 siRNA sequence was correctly cloned to the lentivirus plasmid. Expression levels of IL-6 mRNA and protein and the secretion level of IL-6 significantly decreased after MSCs were infected by the siIL-6 lentivirus. After being separated and co-cultured with siIL-6-MSCs, the apoptosis of OGD injured PC12 cells increased; protein expression levels of STAT3 and Bcl-2 significantly decreased; and the protein expression of Bax significantly increased. Conclusion MSCs-originated IL-6 may regulate the anti-apoptotic function of OGD injured PC12 cells through the STAT3 signaling pathway.

Key words: mesenchymal stem cells, interleukin-6, oxygen glucose deprivation, nerve injury, STAT3, anti-apoptosis