Exosomes derived from human umbilical cord mesenchymal stem cells attenuate edema of spinal astrocytes after oxygen-glucose deprivation/reoxygenation injury in rats

doi:10.3969/j.issn.2095-4344.1788

Abstract

Abstract:

BACKGROUND: Exosomes can regulate and repair nerve cells, thus promoting the proliferation of nerve cells to alleviate spinal cord injury. Exosomes have been less reported to alleviate edema after spinal cord injury.
OBJECTIVE: To study the effect and mechanism by which human umbilical cord mesenchymal stem cell-derived exosomes (hucMSCs-exo) alleviate the edema of spinal astrocytes induced by oxygen-glucose deprivation/reoxygenation.
METHODS: Ultrahigh speed centrifugation method was used to isolate and extract hucMSCs-exo. Spinal astrocytes were extracted from newborn Sprague-Dawley rats by trypsin digestion. Part One: The cells were (1) cultured normally in normal control group, (2) subjected to 6 hours of oxygen-glucose deprivation and 24 hours of reoxygenation in model group, or (3) cultured in medium containing 30, 60, and 90 μg of hucMSCs-exo for 24 hours following 6 hours of oxygen-glucose deprivation and 24 hours of reoxygenation in hucMSCs-exo group. Western blot assay was performed to detect the expression of AQP4. Living cell workstation was used to detect cell volume. Transmission electron microscope was used to observe the ultrastructure of intracellular edema. Part Two: The cells were (1) cultured normally in normal control group, (2) subjected to 6 hours of oxygen-glucose deprivation and 24 hours of reoxygenation in model group, (3) cultured in medium containing 90 μg of hucMSCs-exo for 24 hours following 6 hours of oxygen-glucose deprivation and 24 hours of reoxygenation in hucMSCs-exo group, or (4) cultured in medium containing 5 μmol/L JNK inhibitor sp600125 for 24 hours following 6 hours of oxygen-glucose deprivation and 24 hours of reoxygenation in JNK inhibitor sp600125 group. AQP4 expression and p-JNK protein expression were measured by western blot assay. Living cell workstation was used to detect cell volume
RESULTS AND CONCLUSION: (1) After 24-hour hucMSCs-exo intervention, mitochondrial and endoplasmic reticular edemas were alleviated, the number of lysosomes decreased, the volume of intracellular cell edema was significantly reduced, and the expression of AQP4 in the cells was significantly lowered as compared with the model group (P < 0. 05). (2) Intervention with hucMSCs-exo and JNK inhibitor SP600125 significantly reduced the expression of AQP4 and p-JNK (P < 0.05) and the volume of intracellular edema (P < 0.05) as compared with the model group. Therefore, hucMSCs-exo can reduce the expression of AQP4 protein in spinal astrocytes after oxygen-glucose deprivation/reoxygenation by inhibiting JNK signaling pathway, thereby alleviating cell edema.

Key words: human umbilical cord mesenchymal stem cell-derived exosomes, astrocytes, spinal cord injury, oxygen-glucose deprivation/reoxygenation injury, intracellular edema, AQP4, JNK receptor inhibitor

CLC Number:

Zhang Yong, Ma Xun, Sun Lin, Zhang Li, Guan Xiaoming, Lü Cong, Chen Xu. Exosomes derived from human umbilical cord mesenchymal stem cells attenuate edema of spinal astrocytes after oxygen-glucose deprivation/reoxygenation injury in rats[J]. Chinese Journal of Tissue Engineering Research, 2019, 23(25): 4011-4017.

Figures/Tables 6

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