Research on the mechanism of high glucose affecting the apoptosis of schwann cells by Nox4 NADPH oxidase

doi:10.12047/j.cjap.5719.2019.029

Abstract

Abstract: Objective: To investigate the mechanism of high glucose affecting the apoptosis of schwann cells through Nox4 NADPH oxidase. Methods: The schwann cells of newborn Wistar rats were cultured in vitro. The cultured cells were divided into four groups: control group, high-glucose group, NOX4 siRNA group and control siRNA group (n=10). The WST-1 method was used to detect the cell vitality, and the DCFH-DA method was used to detect the contents of intracellular reactive oxygen free radicals (ROS). Nox4 and Caspase3 mRNA expressions were detected by real-time fluorescence quantitative RT-PCR. Nox4 and Caspase3 protein expressions were determined by Western blot. Results: High glucose culture up-regulated Nox4 mRNA and protein expressions of schwann cells, decreased activity of schwann cells, increased intracellular ROS content, and promoted apoptosis by increasing Caspase3 mRNA and protein expressions. NOX4 siRNA blocked the accumulation of ROS in the high glucose cultured schwann cells, and reduced the damage of glucose on cell viability, by inhibiting NOX4 gene expression. NOX4 siRNA also reduced cell apoptosis by down-regulating Caspase3 mRNA and protein expressions. Conclusion: Nox4 was involved in the hyperglycemic-induced apoptosis of schwann cells through ROS. The regulation of Nox4 expression or function might be a new way to treat diabetic peripheral neuropathy.

Key words: diabetes, schwann cells, NADPH oxidase, reactive oxygen free radicals, apoptosis

YU Ting , XIN Qing , XU Fei, LI Lei. Research on the mechanism of high glucose affecting the apoptosis of schwann cells by Nox4 NADPH oxidase[J]. CJAP, 2019, 35(2): 130-134.

References

[1] Feldman EL, Nave KA, Jensen TS, et al. New horizons in diabetic neuropathy: mechanisms, bioenergetics, and pain[J]. Neuron, 2017, 93(6): 1296-1313.
[2] Yu T, Li L, Bi Y, et al. Erythropoietin attenuates oxidative stress and apoptosis in Schwann cells isolated from streptozotocin-induced diabetic rats [J]. J Pharm Pharmacol, 2014, 66(8): 1150-1160.
[3] 陈娟, 崔节达, 郭晓桐, 等. 慢性阻塞性肺疾病小气道 NOX4、TGF -β的表达与气道重塑的关系[J]. 中国应用生理学杂志, 2017, 33(6): 481-487.
[4] Nisimoto Y, Diebold BA, Cosentino-Gomes D, et al. Nox4: a hydrogen peroxide-generating oxygen sensor [J]. Biochemistry, 2014, 53(31): 5111-5120.
[5] Gon alves NP, V gter CB, Pallesen LT. Peripheral glial cells in the development of diabetic neuropathy [J]. Front Neurol, 2018, 9: 268.
[6] 马益梅, 李传达, 朱雅冰, 等. RvD1对大鼠2型糖尿病神经病理性痛的作用及机制研究[J]. 中国应用生理学杂志, 2017, 33(3), 277-281.
[7] Lennertz RC, Medler KA, Bain JL, et al. Impaired sensory nerve function and axon morphology in mice with diabetic neuropathy [J]. J Neurophysiol, 2011, 106(2): 905-914.
[8] Volpe CMO, Villar-Delfino PH, Dos Anjos PMF, et al. Cellular death, reactive oxygen species (ROS) and diabetic complications [J]. Cell Death Dis, 2018, 9(2): 119.
[9] Zhang L, Pang S, Deng B, et al. High glucose induces renal mesangial cell proliferation and fibronectin expression through JNK/NF-κB/NADPH oxidase/ROS pathway, which is inhibited by resveratrol [J]. Int J Biochem Cell Biol, 2012, 44(4): 629-638.
[10]何婷. 白藜芦醇抑制糖尿病肾病肾间质纤维化的作用和机制研究 [D]. 重庆: 第三军医大学, 2015: 75-76.
[11]Rozentsvit A, Vinokur K, Samuel S, et al. Ellagic acid reduces high glucose-induced vascular oxidative stress through ERK1/2/NOX4 signaling pathway [J]. Cell Physiol Biochem, 2017, 44(3): 1174-1187.
[12]Li Q, Lin Y, Wang S, et al. GLP-1 inhibits high-glucose-induced oxidative injury of vascular endothelial cells [J]. Sci Rep, 2017, 7(1): 8008.
[13]Yao M, Cao F, Wang X, et al. Nox4 is involved in high glucose-induced apoptosis in renal tubular epithelial cells via Notch pathway [J]. Mol Med Rep, 2017, 15(6): 4319-4325.
[14]Hou Q, Lei M, Hu K, et al. The effects of high glucose levels on reactive oxygen species-induced apoptosis and involved signaling in human vascular endothelial cells [J]. Cardiovasc Toxicol, 2015, 15(2): 140-146.
[15]Jha JC, Gray SP, Barit D, et al. Genetic targeting or pharmacologic inhibition of NADPH oxidase nox4 provides renoprotection in long-term diabetic nephropathy [J]. J Am Soc Nephrol, 2014, 25(6): 1237-1254.