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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

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