Chinese Journal of Tissue Engineering Research ›› 2024, Vol. 28 ›› Issue (31): 4921-4925.doi: 10.12307/2024.708

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Effect of Pax6 gene expression on hydrogen peroxide-induced aging in bone marrow mesenchymal stem cells

Gao Jie1, 2, 3, Zou Xingxing1, Wen Banghong1, Li Yuandi1, Su Min1, 3, 4, Hu Rong1, 2, 3   

  1. 1Department of Histology and Embryology, School of Basic Medicine, Guizhou Medical University, Guiyang 550025, Guizhou Province, China; 2Translational Medicine Research Center, Guizhou Medical University, Guiyang 550025, Guizhou Province, China; 3Key Laboratory of Autoimmune Disease Research in Guizhou Colleges and Universities, Guiyang 550025, Guizhou Province, China; 4National and Guizhou Joint Engineering Laboratory for Cell Engineering and Biomedicine Technique, Guizhou Province Key Laboratory of Regenerative Medicine, Guizhou Medical University, Key Laboratory of Adult Stem Cell Transformation of Chinese Academy of Medical Sciences, Guiyang 550025, Guizhou Province, China
  • Received:2023-08-23 Accepted:2023-10-12 Online:2024-11-08 Published:2024-01-22
  • Contact: Hu Rong, MD, Professor, Department of Histology and Embryology, School of Basic Medicine, Guizhou Medical University, Guiyang 550025, Guizhou Province, China; Translational Medicine Research Center, Guizhou Medical University, Guiyang 550025, Guizhou Province, China; Key Laboratory of Autoimmune Disease Research in Guizhou Colleges and Universities, Guiyang 550025, Guizhou Province, China Su Min, MD, Professor, Department of Histology and Embryology, School of Basic Medicine, Guizhou Medical University, Guiyang 550025, Guizhou Province, China; Key Laboratory of Autoimmune Disease Research in Guizhou Colleges and Universities, Guiyang 550025, Guizhou Province, China; National and Guizhou Joint Engineering Laboratory for Cell Engineering and Biomedicine Technique, Guizhou Province Key Laboratory of Regenerative Medicine, Guizhou Medical University, Key Laboratory of Adult Stem Cell Transformation of Chinese Academy of Medical Sciences, Guiyang 550025, Guizhou Province, China
  • About author:Gao Jie, Master, Senior experimentalist, Department of Histology and Embryology, School of Basic Medicine, Guizhou Medical University, Guiyang 550025, Guizhou Province, China; Translational Medicine Research Center, Guizhou Medical University, Guiyang 550025, Guizhou Province, China; Key Laboratory of Autoimmune Disease Research in Guizhou Colleges and Universities, Guiyang 550025, Guizhou Province, China
  • Supported by:
    National Natural Science Foundation of China, No. 82171343, 82060151 (to SM); Project of Guizhou Science and Technology Department, No. [2020]4Y230 (to SM); Project of Guizhou Science and Technology Department, No. [2022]344 (to GJ); Key Laboratory of Higher Learning Schools of Guizhou Province, No. [2023]016 (to SM)

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Abstract:

BACKGROUND: The occurrence and development of various ophthalmic diseases are closely related to excessive oxidative stress, and the inhibition of oxidative stress response may produce preventive and therapeutic effects. 
OBJECTIVE: To explore the role of Pax6 gene expression on hydrogen peroxide-induced aging of mouse bone marrow mesenchymal stem cells (BM-MSCs). 
METHODS: Resuscitated BM-MSCs, Pax6/BM-MSCs, and shPax6/BM-MSCs were treated with hydrogen peroxide for 24 hours, and then β-galactosidase staining was performed. The proliferation index Ki67 expression and apoptosis were detected by flow cytometry. The expression of senescence-associated molecules (Wnt7a, p21, and p53) was detected by RT-PCR. 
RESULTS AND CONCLUSION: (1) After hydrogen peroxide treatment, the cells of the three groups showed senescence phenotype and β-galactosidase staining was positive. Compared with BM -MSCs group, the expression of positive cells in Pax6/BM-MSCs group was less and that in the shPax6/BM-MSCs group was more, and the difference was statistically significant (P < 0.05). (2) The results of flow cytometry showed that compared with BM-MSCs group, the positive expression of Ki67 in the Pax6/BM-MSCs group increased and the level of apoptosis decreased, while the positive expression of Ki67 decreased and the level of apoptosis increased in the shPax6/BM-MSCs group; the difference was significantly different (P < 0.05). (3) RT-PCR showed that compared with the BM-MSCs group, the expression of Wnt7a, p53, and p21 decreased in the Pax6/BM-MSCs  group, while the expression of Wnt7a, p53, and p21 increased in the shPax6/BM-MSCs group; the difference was significantly different (P < 0.05). (4) These findings indicate that overexpression of Pax6 can antagonize the aging progression of BM-MSCs induced by hydrogen peroxide, which may be related to Wnt signaling pathway.

Key words: bone marrow mesenchymal stem cell, Pax6, Wnt7a, hydrogen peroxide, apoptosis, aging

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