National Immunological Laboratory of Traditional Chinese Medicine Affiliated to Youjiang Medical College for Nationalities, Baise, Guangxi Zhuang Autonomous Region 533000, P.R. China.
Department of Gerontology, Affiliated Hospital of Youjiang Medical University for Nationalities, Baise, Guangxi Zhuang Autonomous Region 533099, P.R. China.
Mol Med Rep. 2025 Jan;31(1). doi: 10.3892/mmr.2024.13371. Epub 2024 Oct 25.
The present study aimed to explore the effect of melittin (MLT) on the growth of Schwann cells (SCs) in high glucose conditions and to understand the mechanisms involved. The goal was to provide a theoretical basis for using MLT in the treatment of diabetic peripheral neuropathy (DPN). The CCK‑8 assay was used to measure cell activity at different concentrations of glucose and MLT. Flow cytometry was employed to analyze the effect of MLT on cell cycle phases and apoptosis in SCs under high glucose conditions. To identify differentially expressed proteins, 4D label‑free quantitative proteomics with liquid chromatography‑mass spectrometry was used, followed by biological analysis to explore potential mechanisms. PCR, western blotting and immunofluorescence were conducted to confirm these mechanisms. Melittin (0.2 g/ml) increased the proliferation of SCs in a high glucose environment. Flow cytometry showed that after MLT treatment, the proportion of cells in the G/M+S phase increased and the combined ratio of early and late apoptosis decreased under high glucose conditions. Proteomics identified 1,784 proteins with significant changes in expression; 725 were upregulated, and 1,059 were downregulated. Kyoto Encyclopedia of Genes and Genomes analysis indicated that the differentially expressed proteins were mainly involved in metabolic pathways and neurodegenerative disease pathways. PCR, western blotting and immunofluorescence confirmed the increase in Crabp2, Wnt3a, C‑Jun, CDK4, CyclinD1 and proliferating cell nuclear antigen. In high glucose conditions, MLT protects SCs from glucose toxicity by upregulating the Crabp2/Wnt/β‑catenin signaling pathway, potentially providing a new treatment for DPN.
本研究旨在探讨蜂毒素（MLT）对高糖条件下施万细胞（SCs）生长的影响，并探讨其相关机制。目的是为 MLT 治疗糖尿病周围神经病变（DPN）提供理论依据。采用 CCK-8 法检测不同浓度葡萄糖和 MLT 下细胞活性。采用流式细胞术分析 MLT 对高糖条件下 SCs 细胞周期和凋亡的影响。采用 4D 无标记定量蛋白质组学结合液相色谱-质谱技术筛选差异表达蛋白，然后进行生物分析以探讨潜在机制。采用 PCR、western blot 和免疫荧光法验证这些机制。蜂毒素（0.2 g/ml）可增加高糖环境中 SCs 的增殖。流式细胞术显示，MLT 处理后，高糖条件下 G/M+S 期细胞比例增加，早期和晚期凋亡的联合比例降低。蛋白质组学鉴定出 1784 种表达有显著变化的蛋白；其中 725 种上调，1059 种下调。京都基因与基因组百科全书分析表明，差异表达蛋白主要参与代谢途径和神经退行性疾病途径。PCR、western blot 和免疫荧光证实 Crabp2、Wnt3a、C-Jun、CDK4、CyclinD1 和增殖细胞核抗原增加。在高糖条件下，MLT 通过上调 Crabp2/Wnt/β-catenin 信号通路来保护 SCs 免受葡萄糖毒性，为 DPN 的治疗提供了新的思路。
