Neural Regeneration Research ›› 2020, Vol. 15 ›› Issue (11): 2154-2161.doi: 10.4103/1673-5374.282271

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Knocking down TRPM2 expression reduces cell injury and NLRP3 inflammasome activation in PC12 cells subjected to oxygen-glucose deprivation

Tao Pan 1, Qiu-Jiao Zhu 2, Li-Xiao Xu 3 , Xin Ding 1 , Jian-Qin Li 4 , Bin Sun 1 , Jun Hua 2, Xing Feng 1   

  1. 1 Department of Neonatology, Children’s Hospital Affiliated to Suzhou University, Suzhou, Jiangsu Province, China
    2 Department of Critical Care Medicine, Children’s Hospital Affiliated to Suzhou University, Suzhou, Jiangsu Province, China
    3 Institute of Pediatrics, Children’s Hospital Affiliated to Suzhou University, Suzhou, Jiangsu Province, China
    4 Blood Section, Children’s Hospital Affiliated to Suzhou University, Suzhou, Jiangsu Province, China
  • Online:2020-11-15 Published:2020-08-23
  • Contact: Jun Hua, huajun_1970@126.com; Xing Feng, MD, feng_xing66@163.com.
  • Supported by:
    This work was supported by the National Natural Science Foundation of China, Nos. 81671532, 81771625 (to XF); the Jiangsu Provincial Key Medical Discipline of China, No. ZDXKA2016013 (to XF); the Jiangsu Provincial Medical Youth Talent of China, No. QNRC2016758 (to XF); the Jiangsu Province Women and Children Health Research Project of China, No. F201750 (to XF); the Public Health Technology Project of Suzhou City of China, No. SYS201765 (to XF); a grant from the Department of Pediatrics Clinical Center of Suzhou City of China, No. Szzx201504 (to XF).

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Abstract: Transient receptor potential melastatin 2 (TRPM2) is an important ion channel that represents a potential target for treating injury caused by cerebral ischemia. However, it is unclear whether reducing TRPM2 expression can help repair cerebral injury, and if so what the mech- anism underlying this process involves. This study investigated the protective effect of reducing TRPM2 expression on pheochromocytoma (PC12) cells injured by oxygen-glucose deprivation (OGD). PC12 cells were transfected with plasmid encoding TRPM2 shRNAS, then subjected to OGD by incubation in glucose-free medium under hypoxic conditions for 8 hours, after which the cells were allowed to reox- ygenate for 24 hours. Apoptotic cells, mitochondrial membrane potentials, reactive oxygen species levels, and cellular calcium levels were detected using flow cytometry. The relative expression of C-X-C motif chemokine ligand 2 (CXCL2), NACHT, LRR, and PYD domain– containing protein 3 (NALP3), and caspase-1 were detected using fluorescence-based quantitative reverse transcription-polymerase chain reaction and western blotting. The rates of apoptosis, mitochondrial membrane potentials, reactive oxygen species levels, and cellular cal- cium levels in the TRPM2-shRNA + OGD group were lower than those observed in the OGD group. Taken together, these results suggest that TRPM2 knockdown reduces OGD-induced neuronal injury, potentially by inhibiting apoptosis and reducing oxidative stress levels, mitochondrial membrane potentials, intracellular calcium concentrations, and NLRP3 inflammasome activation.

Key words: apoptosis, calcium, caspase-1, NLRP3, mitochondrial impairment, oxidative stress, oxygen-glucose deprivation, PC12, shRNA,
TRPM2