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Rutin maintains redox balance to relieve oxidative stress induced by TBHP in nucleus pulposus cells
In Vitro Cellular & Developmental Biology - Animal ( IF 1.5 ) Pub Date : 2021-04-28 , DOI: 10.1007/s11626-021-00581-7
Jian Zhou 1 , Qi Liu 2 , Zhou Yang 3 , Chuhai Xie 2 , Long Ling 2 , Hailan Hu 2 , Yanming Cao 2 , Yan Huang 2 , Yue Hua 4
Affiliation  

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.



中文翻译:

芦丁维持氧化还原平衡以缓解TBHP诱导的髓核细胞氧化应激

芦丁以其抗氧化应激的抗炎和抗氧化特性而闻名。然而,其在髓核细胞 (NPC) 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。从 1 个月大的 SD 大鼠获得初级 NPC。用叔丁基过氧化氢(TBHP)处理NPCs获得氧化应激,并用不同浓度的芦丁观察其对NPCs氧化应激的影响。荧光探针 DCFH-DA 用于检测活性氧化物 (ROS)。通过蛋白质印迹和实时 PCR 检测 NPC 中的抗氧化蛋白和热休克蛋白 70 (HSP70)、锰超氧化物歧化酶 (Mn-SOD)、过氧化氢酶、聚集蛋白聚糖和 II 型胶原蛋白的基因。在TBHP的刺激下,NPCs中ROS的含量显着增加并表现出溶解度相关性。芦丁以剂量依赖性方式有效减少 ROS 的积累。HSP70、Mn-SOD和过氧化氢酶的抗氧化蛋白以及蛋白聚糖和II型胶原的基质蛋白在TBHP的刺激下显着降低,而基质金属蛋白酶13(MMP-13)在TBHP干预后显着增加。芦丁提高了 HSP70、Mn-SOD 和过氧化氢酶的表达,提高了蛋白聚糖和 II 型胶原的含量,并抑制了 NPC 中 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,表明芦丁可能成为椎间盘退变的一种治疗选择。

更新日期:2021-04-29
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