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芦丁维持氧化还原平衡,以减轻叔丁基过氧化氢诱导的髓核细胞氧化应激。

Rutin maintains redox balance to relieve oxidative stress induced by TBHP in nucleus pulposus cells.

作者信息

Zhou Jian, Liu Qi, Yang Zhou, Xie Chuhai, Ling Long, Hu Hailan, Cao Yanming, Huang Yan, Hua Yue

机构信息

Department of Spine and Joint Surgery, Nanchang Hongdu Hospital of TCM, Nanchang, Jiangxi, China.

Department of Orthopaedic Surgery, The Second Affiliated Hospital of Guangzhou Medical University, 250 Chang-gang-dong Road, Guangzhou, 510260, Guangdong, China.

出版信息

In Vitro Cell Dev Biol Anim. 2021 Apr;57(4):448-456. doi: 10.1007/s11626-021-00581-7. Epub 2021 Apr 28.

Abstract

Rutin is well known for its anti-inflammatory and antioxidant properties against oxidative stress. However, its protective function in nucleus pulposus cells (NPCs) remains unclear. This study was aimed to explore the effects of rutin on oxidative stress in NPCs. Primary NPCs were obtained from 1-mo-old SD rats. The NPCs were treated with tert-butyl hydrogen peroxide (TBHP) to obtain the oxidative stress, and different concentrations of rutin were used to observe its influence on the oxidative stress in NPCs. Fluorescent probe DCFH-DA was used to detect reactive oxide species (ROS). The antioxidant proteins and genes of heat shock protein 70 (HSP70), manganese superoxide dismutase (Mn-SOD), catalase, aggrecan and collagen II in NPCs were measured by western blot and real-time PCR. With the stimulation of TBHP, the content of ROS in NPCs increased significantly and showed solubility correlation. Rutin effectively reduced the accumulation of ROS in a dose-dependent manner. The antioxidant proteins of HSP70, Mn-SOD, and catalase and the matrix proteins of aggrecan and collagen II decreased remarkably with the stimulation of TBHP, while the matrix metalloproteinase-13 (MMP-13) significantly increased after TBHP intervention. Rutin boosted the expressions of the HSP70, Mn-SOD, and catalase, elevated the contents of aggrecan and collagen II, and inhibited the expression of MMP-13 in NPCs. The findings of this study suggested that rutin is able to reverse oxidative stress and maintain cellular function of NPCs, and it was indicated that rutin could be a possible therapeutic option for intervertebral disc degeneration.

摘要

芦丁因其抗炎和抗氧化特性以及对抗氧化应激的作用而闻名。然而,其在髓核细胞(NPCs)中的保护功能仍不清楚。本研究旨在探讨芦丁对NPCs氧化应激的影响。原代NPCs取自1月龄的SD大鼠。用叔丁基过氧化氢(TBHP)处理NPCs以诱导氧化应激,并用不同浓度的芦丁观察其对NPCs氧化应激的影响。使用荧光探针DCFH-DA检测活性氧(ROS)。通过蛋白质免疫印迹法和实时定量PCR检测NPCs中热休克蛋白70(HSP70)、锰超氧化物歧化酶(Mn-SOD)、过氧化氢酶、聚集蛋白聚糖和胶原蛋白II的抗氧化蛋白和基因。在TBHP刺激下,NPCs中ROS的含量显著增加且呈剂量相关性。芦丁以剂量依赖的方式有效减少了ROS的积累。在TBHP刺激下,HSP70、Mn-SOD和过氧化氢酶的抗氧化蛋白以及聚集蛋白聚糖和胶原蛋白II的基质蛋白显著降低,而在TBHP干预后基质金属蛋白酶-13(MMP-13)显著增加。芦丁促进了NPCs中HSP70、Mn-SOD和过氧化氢酶的表达,提高了聚集蛋白聚糖和胶原蛋白II的含量,并抑制了MMP-13的表达。本研究结果表明,芦丁能够逆转氧化应激并维持NPCs的细胞功能,提示芦丁可能是椎间盘退变的一种潜在治疗选择。

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