Laboratory of Neuromodulation of Pain, Department of Anatomy, Institute of Biomedical Sciences, University of São Paulo, Av. Professor Lineu Prestes 2415, ICB III, Cidade Universitária, São Paulo, 05508-000, Brazil.
Laboratory of Intracellular Peptides Pharmacology, Department of Pharmacology, University of São Paulo, Av Lineu Prestes 1524, ICB I, São Paulo, 05508-900, Brazil.
Lasers Med Sci. 2020 Feb;35(1):149-156. doi: 10.1007/s10103-019-02808-2. Epub 2019 Jun 20.
Peripheral neuropathy (PN) is a serious complication of diabetes mellitus (DM) and is known to be resistant to conventional treatment. Photobiomodulation (PBM) is demonstrated to be effective in treating PN and in protecting nerve fiber damage. To better understand the mechanisms underlying the regenerative effects of PBM on diabetic neuropathy, we conducted a study in an in vitro model of diabetes induced by glucose neurotoxicity. Neuro 2A cells (1 × 10 cells/ well; N2A) were cultured in Minimum Essential Medium (MEM) supplemented with high glucose concentrations (100 mM) for 48 h and after the incubation period were submitted to either one or three consecutive applications of PBM, once a day (low-level InGaAlP, continuous wave mode, 660 nm, 30 mW, 1.6 J/cm, 15 s, per well). Cell viability was measured by MTT method, neurotoxicity by LDH release, neurite outgrowth was evaluated through morphometric analysis, and AKT/ERK protein expression levels were assessed by western blotting. Results demonstrate that PBM increased N2A viability as well as induced neurogenesis observed by the increase in neurite outgrowth being this effect modulated by AKT activation. Data obtained herein reinforce the regenerative potential of PBM in the treatment of PN and strongly suggests that phototherapy should be considered adjuvant in the treatment of diabetes.
周围神经病变(PN)是糖尿病(DM)的严重并发症,已知对常规治疗具有抗性。光生物调节(PBM)已被证明可有效治疗 PN 并保护神经纤维损伤。为了更好地理解 PBM 对糖尿病性神经病再生作用的机制,我们在葡萄糖神经毒性诱导的体外糖尿病模型中进行了一项研究。将神经 2A 细胞(1×10 个细胞/孔;N2A)在补充有高葡萄糖浓度(100 mM)的最低必需培养基(MEM)中培养 48 h,孵育期后,每天接受一次或连续三次 PBM 处理(低水平 InGaAlP,连续波模式,660 nm,30 mW,1.6 J/cm,15 s,每孔)。通过 MTT 法测量细胞活力,通过 LDH 释放测量神经毒性,通过形态计量分析评估神经突生长,通过 Western blot 评估 AKT/ERK 蛋白表达水平。结果表明,PBM 增加了 N2A 的活力,并通过增加神经突生长来诱导神经发生,这种作用通过 AKT 激活来调节。本文获得的数据增强了 PBM 在治疗 PN 中的再生潜力,并强烈表明光疗应该被考虑作为糖尿病治疗的辅助手段。