Human dental pulp stem cells conditioned medium protects genioglossus myoblast from cobalt chloride-induced hypoxia injury through AMPK/PGC-1α pathway

doi:10.19439/j.sjos.2020.06.003

Abstract

Abstract: PURPOSE: To study the effect of hypoxia induced by cobalt chloride (CoCl₂) on viability and oxidative stress of genioglossus myoblast, and to explore the mechanism of the protective effect of conditioned medium (CM) on human dental pulp stem cells (hDPSCs). METHODS: The hDPSCs were isolated and cultured, and the conditioned medium was prepared by ultrafiltration concentration. Mouse genioglossus myoblasts were isolated and divided into control group, CM group, CoCl₂ group and CoCl₂+CM group. The cell viability of genioglossus myoblasts was detected by CCK-8. The intracellular and mitochondrial ROS levels were evaluated by DCFH-DA and MitoSOX, respectively. The expression level of mitochondria-related genes in NRF-1 and NRF-2 were analyzed by real-time PCR. The expression of PGC-1α, p-AMPK and total AMPK protein was detected by Western blot. Statistical analysis was performed using SPSS 22.0 software package. RESULTS: The proliferation of genioglossus myoblasts was significantly decreased after 200 μmol/L CoCl₂ treatment for 24 h (P<0.05), and the levels of reactive oxygen species (ROS) were significantly increased in intracellular and mitochondria (P<0.05). Compared with CoCl₂ group, the proliferation ability of hDPSCs-CM was dramatically raised (P<0.05), and the intracellular and mitochondrial ROS content was remarkably decreased(P<0.05). hDPSCs-CM up-regulated the protein expression levels of pAMPK and PGC-1α in genioglossus myoblasts and mitochondrial downstream effectors of PGC-1α, including mRNA expression levels of NRF-1, NRF-2 (P<0.05). CONCLUSIONS: Human dental pulp stem cells conditioned medium can alleviate hypoxia injury induced by CoCl₂ in genioglossus myoblasts, and its mechanism may be related to AMPK/PGC-1α signaling pathway.

Key words: Human dental pulp stem cells, Conditioned medium, Genioglossus, Hypoxia, Reactive oxygen species, AMPK/PGC-1α pathway

CLC Number:

R318.01

ZHANG Wei-hua, YU Li-ming, HAN Xin-xin, PAN Jie, DENG Jia-jia, ZHU Lu-ying, LIU Yue-hua. Human dental pulp stem cells conditioned medium protects genioglossus myoblast from cobalt chloride-induced hypoxia injury through AMPK/PGC-1α pathway[J]. Shanghai Journal of Stomatology, 2020, 29(6): 573-579.

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