红景天苷通过抑制MAPK通路和激活Nrf2通路实现脊髓损伤后神经保护的机制研究  

Mechanism Study of Salidroside in Neuroprotection After Spinal Cord Injury by Inhibiting the MAPK Pathway and Activating the Nrf2 Pathway

作　　者：周亚净[1] 段春胜[2] 何润之[3] 程芳[4] 刘建敏[5] Zhou Yajing;Duan Chunsheng;He Runzhi;Cheng Fang;Liu Jianmin(Department of Anesthesiology,Xingtai People's Hospital,Xingtai 054001,China;Department of Pediatric Surgery,Xingtai People's Hospital,Xingtai 054001,China;Third Department of Neurosurgery,Xingtai People's Hospital,Xingtai 054001,China;Department of Dermatology,Xingtai People's Hospital,Xingtai 054001,China;Department of Hand and Foot Surgery,Xingtai People's Hospital,Xingtai 054001,China)

机构地区：[1]邢台市人民医院麻醉科,河北邢台054001 [2]邢台市人民医院小儿外科,河北邢台054001 [3]邢台市人民医院神经外科,河北邢台054001 [4]邢台市人民医院皮肤科,河北邢台054001 [5]邢台市人民医院手足外科,河北邢台054001

出　　处：《南开大学学报（自然科学版）》2024年第1期55-60,共6页Acta Scientiarum Naturalium Universitatis Nankaiensis

基　　金：河北省青年拔尖人才支持项目。

摘　　要：脊髓损伤(pinal cord injury,SCI)是世界范围内的一个公共卫生问题.SCI通常会引发过度炎症反应,导致继发性组织损伤和进一步的细胞和器官功能障碍.除了炎症反应,氧化应激是SCI继发性损伤的重要组成部分.因此,减少炎症和氧化应激是治疗SCI的策略.目的是研究红景天苷(SAD)是否分别通过抑制丝裂原活化蛋白激酶(MAPK)途径和激活Nrf2通路对SCI发挥抗炎和抗氧化作用.在体外实验中,使用q RT-PCR法和酶联免疫吸附实验(ELISA)检测不同浓度SAD对脂多糖(LPS)激活的星形胶质细胞(AS)中白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)和肿瘤坏死因子-α(TNF-α)表达以及不同浓度SAD对过氧化氢(H_(2)O_(2))诱导的星形胶质细胞各组细胞中MDA含量、ROS水平、SOD和GSH-Px活力;q RT-PCR法和western blot分别检测AS中P38、p-P38、JNK、p-JNK、ERK、p-ERK以及Nrf2、HO-1和NQO-1的表达.结果表明,不同浓度SAD抑制了对脂多糖(LPS)激活的星形胶质细胞中白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)和肿瘤坏死因子-α(TNF-α)表达和释放.不同浓度的红景天苷能够通过减少MDA含量和ROS水平同时提高SOD和GSH-Px活力.此外,通过q RT-PCR和蛋白质印迹分析发现SAD抑制了P38、JNK和细胞外调节蛋白激酶(ERK)的表达并上调了Nrf2、HO-1、NQO-1的表达.结论是SAD可以通过抑制MAPK信号通路中P38、JNK和ERK磷酸化,减少SCI后小鼠星形胶质细胞的炎症反应,并通过激活Nrf2信号通路中Nrf2、HO-1、NQO-1减轻H_(2)O_(2)诱导的AS氧化损伤.Spinal cord injury(SCI) is a public health issue worldwide.SCI typically triggers an excessive inflammatory response,leading to secondary tissue damage and further cellular and organ dysfunction.In addition to inflammatory response,oxidative stress is an important component of secondary injury in SCI.Therefore,reducing inflammation and oxidative stress has great potential for therapeutic strategies for SCI.In this study,the aim was to investigate whether salidroside (SAD) exerts anti-inflammatory and antioxidant effects on SCI by inhibiting the mitogen-activated protein kinase (MAPK) pathway and activating the Nrf2 pathway,respectively.Detection of IL-1β,IL-6,and TNF-α expression in astrocytes (AS) activated by lipopolysaccharide (LPS)at different concentrations of SAD using enzyme-linked immunosorbent assay (ELISA) in vitro and different concentrations of SAD on MDA content,ROS level,SOD and GSH-Px activity in AS induced by hydrogen peroxide (H_2O_2).Reverse transcription polymerase chain reaction (RT-PCR) and western blotting were used to detect the expression of P38,p-P38,JNK,p-JNK,ERK,p-ERK,Nrf2,HO-1,and NQO-1 in AS,respectively.The results showed that different concentrations of SAD inhibited IL-1β,IL-6,and TNF-αexpression and release in AS activated by LPS.Different concentrations of SAD can increase SOD and GSH-Px activity and reduce MDA content and ROS levels.In addition,RT-PCR and western blot analysis revealed that SAD inhibits the expression of p38,JNK,and ERK,as well as upregulates the expression of Nrf2,HO-1,and NQO-1.Overall,SAD can reduce inflammation in AS activated by LPS by inhibiting phosphorylation of P38,JNK,and ERK in the MAPK signaling pathway,and alleviate H_2O_2-induced oxidative damage in AS by activating Nrf2,HO-1,and NQO-1 in the Nrf2 signaling pathway.

关 键 词：星形胶质细胞 炎症 氧化应激 红景天苷 脊髓损伤 

分 类 号：R68[医药卫生—骨科学]