Yang Xinwei, Yao Weijie, Shi Haotian, Liu Haolong, Li Yangfan, Gao Yanbin, Liu Renhui, Xu Liping
School of Traditional Chinese Medicine, Capital Medical University, #10, Youanmenwai Xitoutiao, Fengtai District, Beijing 100069, China; Beijing Key Lab of TCM Collateral Disease Theory Research China.
School of Traditional Chinese Medicine, Capital Medical University, #10, Youanmenwai Xitoutiao, Fengtai District, Beijing 100069, China.
J Ethnopharmacol. 2016 Jun 5;185:361-9. doi: 10.1016/j.jep.2016.03.031. Epub 2016 Mar 12.
Paeoniflorin (PF) is the principal bioactive component of Paeonia lactiflora Pall., which an included in Tang Luo Ning recipe, a traditional Chinese herbal medicine based on Huangqi Guizhi Wuwu decoction. PF is also widely used in Traditional Chinese Medicine for the treatment of blood-arthralgia disease including diabetic peripheral neuropathy (DPN), but its underlying molecular mechanism of neuroprotective effects is not yet well understood. Diabetic hyperglycemia induced oxidative stress in Schwann cells, an important component of the peripheral nervous system, has been proposed as a unifying mechanism for DPN. The objective of this study is to determine the effects of PF on Schwann cells oxidative stress and apoptosis induced by high glucose.
RSC96 cells, a Schwann cell line, were treated with high glucose (150mM) and PF (1, 10 and 100μM). Subsequently, MTT assay was performed. The level of apoptosis was examined by flow cytometry and the oxidative stress was reflected by reactive oxygen species (ROS), malondialdehyde (MDA), glutathione S-transferases (GST) and glutathione peroxidase (GPX) levels. The mRNA expressions of Nuclear factor-E2-related factor 2 (Nrf2) and heme oxygenase 1 (HO-1) were detected by qRT-PCR. The levels of Kelch-like ECH-associating protein 1 (Keap1), Nrf2, HO-1, γ-glutamylcysteine synthetase (γGCS), B-cell CLL/lymphoma 2 (Bcl-2), Bax and Caspase 3 were detected by High content analysis and/or Western blot.
The role of PF markedly suppressed high glucose induced Schwann cells oxidative stress by decreasing ROS and MDA levels and increasing GST and GPX activity. Western blot analysis showed that PF induced nuclear translocation of Nrf2. High content analysis showed that PF promoted Nrf2 dissociation from Keap1 and upregulating the Nrf2/ antioxidant response element (ARE) pathway. Furthermore, PF reduced Schwann cells apoptosis by increasing Bcl-2 and inhibiting Bax and Caspase-3 expressions.
PF in the management of Schwann cells oxidative stress induced by high glucose may be associated with activation of Nrf2/ARE pathway and Bcl-2-related apoptotic pathway.
芍药苷（PF）是芍药的主要生物活性成分，芍药被纳入糖络宁方中，糖络宁方是一种基于黄芪桂枝五物汤的传统中药。PF在传统中医中也广泛用于治疗包括糖尿病周围神经病变（DPN）在内的血痹病，但其神经保护作用的潜在分子机制尚未完全明确。糖尿病高血糖诱导外周神经系统重要组成部分雪旺细胞的氧化应激，已被认为是DPN的一个统一机制。本研究的目的是确定PF对高糖诱导的雪旺细胞氧化应激和凋亡的影响。
雪旺细胞系RSC96细胞用高糖（150mM）和PF（1、10和100μM）处理。随后，进行MTT试验。通过流式细胞术检测凋亡水平，活性氧（ROS）、丙二醛（MDA）、谷胱甘肽S-转移酶（GST）和谷胱甘肽过氧化物酶（GPX）水平反映氧化应激。通过qRT-PCR检测核因子E2相关因子2（Nrf2）和血红素加氧酶1（HO-1）的mRNA表达。通过高内涵分析和/或蛋白质免疫印迹法检测 Kelch样ECH相关蛋白1（Keap1）、Nrf2、HO-1、γ-谷氨酰半胱氨酸合成酶（γGCS）、B细胞淋巴瘤/白血病-2（Bcl-2）、Bax和半胱天冬酶3的水平。
PF的作用显著抑制高糖诱导的雪旺细胞氧化应激，降低ROS和MDA水平，增加GST和GPX活性。蛋白质免疫印迹分析表明PF诱导Nrf2核转位。高内涵分析表明PF促进Nrf2与Keap1解离并上调Nrf2/抗氧化反应元件（ARE）途径。此外，PF通过增加Bcl-2和抑制Bax及半胱天冬酶-3表达来减少雪旺细胞凋亡。
PF对高糖诱导的雪旺细胞氧化应激的调控作用可能与激活Nrf2/ARE途径和Bcl-2相关凋亡途径有关。
