Journal of Jilin University Medicine Edition ›› 2016, Vol. 42 ›› Issue (03): 430-434.doi: 10.13481/j.1671-587x.20160303

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Effect of autophagy inducedby hypoxia on cell proliferation in olfactory ensheathing cells from olfactory mucosa of rats

WANG Jun1, ZHANG Lijian1, XIA Hechun2, LIANG Xueyun3, WANG Yaping1   

  1. 1. Department of Clinical Medicine, College of Clinical Medical Sciences, Ningxia Medical University, Yinchuan 750004, China;
    2. Department of Neurosurgery, General Hospital, Ningxia Medical University, Yinchuan 750004, China;
    3. Institute of Human Stem Cells, General Hospital, Ningxia Medical University, Yinchuan 750004, China
  • Received:2015-12-02 Published:2016-06-17

Abstract:

Objective: To discuss the occurrence of hypoxia-induced autophagy in the olfactory ensheathing cells (OECs) from olfactory mucosa in the newly-born SD rat models,and to evaluate the effect of autophagy on the cell proliferation. Methods: The OECs were obtained and cultivated by enzyme digestion and advanced Nash differential attachment,and the properties and purities of OECs were identified by immunofluorescent assay;the OECs cultured in hypoxia condition with 2% oxygen levels were used as hypoxia group,the cells cultured under the normal culture conditions were regarded as control group, and the OECs treated with autophagy inhibitor 3-MA and cultured under hypoxic conditions were regarded as 3-MA+hypoxia group.The morphology of cells in various groups was observed under phase contrast microscope; Western blotting method was used to measure the protein expression levels of hypoxia inducible factor-1(HIF-1α ),LC3Ⅰ/Ⅱ (autophagy marker protein) and Beclin-1 (autophagy marker gene) in the cells in control group,4 h hypoxia group,and 8 h hypoxia group;CCK-8 method was used to measure the influence of autophagy in cell proliferation in control group, 3-MA group,hypoxia group and 3-MA+hypoxia group, Results: After separated and purificated,the OECs in control group,hypoxia group and 3-MA+hypoxia group showed fusiform, typical bipolar,and tripolar cell morphology,and had no significant differences.In the cultivated OECs,the NGFRp 75 and GFAP were expressed positively through separation and purification,and the purity was 87%. Compared with control group,the expression level of HIF-1α in the OECs in hypoxia group was increased (P<0.05),and the expression levels of LC3I Ⅱ and Beclin-1 in the OECs in hypoxia group were increased (P<0.05).Compared with hypoxia group,the proliferation ability of OECs in 3-MA+hypoxia group was reduced. Conclusion: The autophagy induced by hypoxia can decrease the cell proliferation of OECs from olfactory mucosa in the newly-born SD rats.

Key words: olfactory ensheathing cells, hypoxia, autophagy, cell proliferation

CLC Number: 

  • R329.28