Vitamin B1 and B12 mitigates neuron apoptosis in cerebral palsy by augmenting BDNF expression through MALAT1/miR-1 axis - PubMed (original) (raw)

Vitamin B1 and B12 mitigates neuron apoptosis in cerebral palsy by augmenting BDNF expression through MALAT1/miR-1 axis

En-Yao Li et al. Cell Cycle. 2019 Nov.

Abstract

Through the roles of vitamin B1 and B12 in neuroprotection and in improving cerebral palsy symptoms have been previously noticed, the action mechanism is still unclear. This study aims to investigate the protective effect of vitamin B1 and B12 on neuron injury in cerebral palsy and to clarify the mechanism of vitamin B1 and B12 inhibiting neurons apoptosis, and to focus on the role of lncRNA MALAT1 in this process. In order to investigate the effect of vitamin B1 and B12 on neurons injury in vivo and on neuron apoptosis in vitro, we, respectively, introduced vitamin B1 and B12 into cerebral palsy rat and in apoptosis-induced N2A neurons by Oxygen Glucose Deprivation/reoxygenation (OGD/R). Our results demonstrated that vitamin B1 and B12 treatment improved the motor and memory functions and ameliorated the neurons injury in cerebral palsy rats. OGD/R treatment repressed the expression of MALAT1 and BDNF and the phosphorylation of PI3K and Akt, and enhanced the miR-1 expression, which were all reversed by vitamin B1 and B12 treatment in N2A neurons. Vitamin B1 and B12 inhibited miR-1 expression through MALAT1, promoted BDNF expression and activated PI3K/Akt signaling through the MALAT1/miR-1 axis. Vitamin B1 and B12 suppressed neuron apoptosis by up-regulating BDNF via MALAT1/miR-1 pathway. MALAT1 interference abolished the neuroprotective effect of vitamin B1 and B12 in cerebral palsy rats. Collectively, vitamin B1 and B12 up-regulates BDNF and its downstream PI3K/Akt signaling through MALAT1/miR-1 axis, thus suppressing neuron apoptosis and mitigating nerve injury in cerebral palsy rats.

Keywords: BDNF; Cerebral palsy; MALAT1; miR-1; vitamin B1 and B12.

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Figures

Figure 1.

Figure 1.

Vitamin B1 and B12 introduced by HA ameliorated nerve injury in cerebral palsy rats. Rats were allocated into three groups (n = 6): Sham, CP, CP+HA. (a) The motor and memory functions of rats were evaluated by Rota-rod test and Step-down avoidance task, respectively; the NSE level was detected by ELISA. (b) The expression of lncRNAs including MALAT1 in hippocampus tissues was quantified by qRT-PCR. (c) The expression level of miR-1, BDNF mRNA and protein in hippocampus tissues was analyzed by qRT-PCR and western blot, respectively. *P < 0.05 compared with sham; #P < 0.05 compared with CP.

Figure 2.

Figure 2.

The expression of MALAT1 in OGD/R induced N2A neurons was increased by vitamin B1 and B12. To evaluate the effect of vitB on MALAT1 expression and which levels of vitB were suitable in N2A cells, OGD/R and VitB at different concentrations were introduced to allocate into six groups: control, OGD/R, OGD/R + 1 μM VitB, OGD/R + 2.5 μM VitB, OGD/R + 5 μM VitB, and OGD/R + 7.5 μM VitB. (a) The expression of MALAT1 in N2A cells was detected by qRT-PCR. (b, c) The expression level of miR-1, BDNF mRNA and protein in N2A cells was determined by qRT-PCR and western blot, respectively. *P < 0.05 compared with control; #P < 0.05 compared with OGD/R; &P < 0.05 compared with 1μM; $P < 0.05 compared with 2.5μM.

Figure 3.

Figure 3.

Vitamin B1 and B12 reduced miR-1 expression in OGD/R induced N2A neurons through up-regulating MALAT1. (a) To evaluate which miRNA was affected by vitB, N2A cells were assigned into three groups: control, OGD/R, OGD/R+ VitB (5μM). The level of miR-140, miR-141, miR-142, miR-150, miR-155, and miR-1 in N2A cells was evaluated using qRT-PCR. (b) N2A cells were then divided into five groups via different treatments: control, OGD/R, OGD/R+ VitB (5μM), OGD/R+ VitB+si-control, and OGD/R+ VitB+si-MALAT1. The expression of miR-1 in N2A cells of different groups was examined by qRT-PCR. *P < 0.05 compared with control; #P < 0.05 compared with OGD/R; &P < 0.05 compared with the OGD/R+ VitB+si-control group.

Figure 4.

Figure 4.

Vitamin B1 and B12 affected BDNF and PI3K/Akt pathway through MALAT1/miR-1 axis. N2A cells were allocated into seven groups: control, OGD/R, OGD/R+ VitB (5μM), OGD/R+ VitB+si-control, OGD/R+ VitB+si-MALAT1 (20μM), OGD/R+ VitB+si-MALAT1+ NC, and OGD/R+ VitB+si-MALAT1+ miR-1 inhibitor. (a) The expression of miR-1 in N2A cells of different groups was assessed with qRT-PCR. (b) The protein level of p-PI3K, PI3K, p-Akt, Akt, and BDNF in N2A cells was determined using western blot. *P < 0.05 compared with control; #P < 0.05 compared with OGD/R; &P < 0.05 compared with the OGD/R+ VitB+si-control group; $P < 0.05 compared with the OGD/R+ VitB+si-MALAT1+ NC group.

Figure 5.

Figure 5.

Vitamin B1 and B12 participated in neuron apoptosis via the MALAT1/miR-1/BDNF/PI3K/Akt pathway. N2A cells were grouped according to different treatments: control, OGD/R, OGD/R+ VitB (5μM), OGD/R+ VitB+si-control, OGD/R+ VitB+si-MALAT1, OGD/R+ VitB+si-MALAT1+ NC, OGD/R+ VitB+si-MALAT1+ miR-1 inhibitor, and OGD/R+ VitB+si-MALAT1+ miR-1 inhibitor+LY294002 (inhibitor of PI3K/Akt pathway). (a) Neuron apoptosis was estimated using TUNEL staining. (b) The level of cleaved-capase3 was analyzed by western blot. *P < 0.05 compared with control; #P < 0.05 compared with OGD/R; &P < 0.05 compared with the OGD/R+ VitB+si-control group; $P < 0.05 compared with the OGD/R+ VitB+si-MALAT1+ NC group; %P < 0.05 compared to the OGD/R+ VitB+si-MALAT1+ miR-1 inhibitor group.

Figure 6.

Figure 6.

Down-regulation of MALAT1 abolished the neuroprotective effect of HA-introduced vitamin B1 and B12 in cerebral palsy rats. Rats were randomly allocated into several groups (n = 6 in each group): Sham, CP, CP+HA-VitB, CP+HA+si-control, and CP+HA+si-MALAT1. (a) The motor and memory functions of rats were evaluated by Rota-rod test and Step-down avoidance task, respectively; the NSE level was detected by ELISA. (b) The expression of MALAT1, miR-1, and BDNF mRNA in hippocampus tissues was quantified with qRT-PCR; the level of BDNF protein in hippocampus tissues was analyzed by western blot. *P < 0.05 compared with sham; #P < 0.05 compared with CP.

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References

    1. Mathewson MA, Lieber RL.. Pathophysiology of muscle contractures in cerebral palsy. Phys Med Rehabil Clin N Am. 2015;26(1):57–67. - PMC - PubMed
    1. Rumajogee P, Bregman T, Miller SP, et al. Rodent hypoxia-ischemia models for cerebral palsy research: a systematic review. Front Neurol. 2016;7:57. - PMC - PubMed
    1. Ding ZM, Wu B, Zhang WQ, et al. Neuroprotective effects of ischemic preconditioning and postconditioning on global brain ischemia in rats through the same effect on inhibition of apoptosis. Int J Mol Sci. 2012;13(5):6089–6101. - PMC - PubMed
    1. Gelderblom M, Daehn T, Schattling B, et al. Plasma levels of neuron specific enolase quantify the extent of neuronal injury in murine models of ischemic stroke and multiple sclerosis. Neurobiol Dis. 2013;59:177–182. - PubMed
    1. Wang JL, Li RL, Jia CS. Establishment of data warehouse of needling and moxibustion literature based on data mining. Zhen Ci Yan Jiu. 2012;37(1):67–71. - PubMed

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This study was supported by the grants from the Key Scientific Research Project of Colleges and Universities in Henan province [Grant No. 14A360016], Zhengzhou Collaborative Innovation Major Project [Grant No. 18XTZX12003], Special Research Project of Traditional Chinese Medicine in Henan Province [Grant No. 2018ZY3013], Henan medical science and technology project [Grant No. 2018020223], Key Technological Research Projects of Henan Science and Technology Department [Grant No. 192102310366], Henan Science and Technology Development Plan Project [Grant No. 172102310170], Key Scientific Research Projects Plan of Henan Higher Education Institutions [Grant No. 17A320007], Key Scientific Research Projects Plan of Henan Higher Education Institutions [Grant No. 16B320026], Key Scientific Research Projects Plan of Henan Higher Education Institutions [Grant No. 14A360016], Research Project on Teaching Reform of Vocational Education in Henan Province [Grant No. ZJC18051].

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