Effects of microRNA-99a on Neuro-2a cells against oxidative injury induced by hydrogen peroxide

doi:10.3969/j.issn.1006-7795.2014.03.009

Abstract

Abstract:

Objective To investigate the effects of microRNA-99a(miR-99a) on neuro-2a cells against oxidative injury induced by hydrogen peroxide(H₂O₂). Methods Neuro-2a cells were divided into 3 groups by different treatments: control group, control+H₂O₂(100 μmol/L) group and miR-99a mimics+H₂O₂ group. Cell viability was measured by cell counting kit-8. Total superoxide dismutase(T-SOD) activity, manganese superoxide dismutase(Mn-SOD) activity, and content of reduced nicotinamide adenine dinucleotide(NADH) were assayed by a variety of biochemical kits according to the instructions respectively. Protein expression levels of synaptosoma associated protein of molecular mass 25 000(SNAP25), Mn-SOD and extracellular SOD(EC-SOD) were assessed by Western blotting. Results Compared with control group, cell viability of neuro-2a in the control+H₂O₂ group and miR-99a+H₂O₂ group, decreased significantly to 85% and 89% respectively upon hydrogen peroxide stimulation(P<0.05). However, miR-99a+H₂O₂ group appeared to have less reduction of 11% than control+H₂O₂ group(P<0.05). Further studies showed that enzyme activities of T-SOD and Mn-SOD were depressed in control+H₂O₂ group in comparison with control group(P<0.05). Whereas, with pretreatment of miR-99a mimics transfection, T-SOD and Mn-SOD activities were enhanced to a significantly higher level, relative to the normal level of control group(P<0.05), as well as greatly increased protein expression of Mn-SOD and EC-SOD, in contrast to control group and control+H₂O₂ group(P<0.05). Besides, NADH content of neuro-2a in control+H₂O₂ group was reduced compared with control group(P<0.05), while in miR-99a+H₂O₂ group, NADH content exceeded the normal level of control group(P<0.05). It was also found that protein expression of SNAP25 increased in the two groups of hydrogen peroxide stimulation. And miR-99a+H₂O₂ group had a tendency of more increase in protein expression of SNAP25, compared to control+H₂O₂ group. Conclusion miR-99a can effectively protect neuro-2a cells from oxidative injury induced by hydrogen peroxide.

Key words: miR-99a, neuro-2a, H₂O₂, oxidative injury

CLC Number:

R743.9

Tao Zhen, Wang Rongliang, Zhao Haiping, Luo Yumin. Effects of microRNA-99a on Neuro-2a cells against oxidative injury induced by hydrogen peroxide[J]. Journal of Capital Medical University, 2014, 35(3): 305-309.

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URL: https://journal03.magtech.org.cn/Jweb_sdykdxxb/EN/10.3969/j.issn.1006-7795.2014.03.009

https://journal03.magtech.org.cn/Jweb_sdykdxxb/EN/Y2014/V35/I3/305

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