Chen Yanke, Zhang Li, Ni Jingshu, Wang Xiaoyu, Cheng Jian, Li Yuanchao, Zhen Xuechu, Cao Ting, Jia Jia
Jiangsu Key Laboratory of Translational Research and Therapy for Neuro-Psycho-Diseases and College of Pharmaceutical Sciences, Soochow University, PR China.
Jiangsu Key Laboratory of Translational Research and Therapy for Neuro-Psycho-Diseases and Institute of Neuroscience, Soochow University, Suzhou, Jiangsu, 215123, PR China.
J Pharmacol Sci. 2016 Jun;131(2):131-7. doi: 10.1016/j.jphs.2016.05.003. Epub 2016 May 20.
(5R)-5-Hydroxytriptolide (LLDT-8), an analogue of triptolide, displays lower toxicity compared to triptolide and has comparable immunosuppressive effects. We investigated the anti-inflammatory and neuroprotective effects of LLDT-8 on cerebral ischemia/reperfusion injury.
Nitric oxide production from microglia was assessed by measuring the nitrite concentration in the culture medium with Griess reagent. Microglial cells and ischemic brain tissues were examined for the expression of proinflammatory mediators by qPCR and western blot. Infarct volumes were assessed with TTC histology. The TLR4/NF-κB signaling pathway was analyzed with western blot and immunocytochemistry.
LLDT-8 significantly reduced infarct sizes and expression of pro-inflammatory cytokines in the ischemic cortex. LLDT-8 inhibited NO release and expression of TNF-α, IL-1β and iNOS in BV-2 microglia and primary microglia treated with LPS. In addition, LLDT-8 suppressed expression of TLR4, degradation of IκBα and nuclear translocation of NF-κB.
LLDT-8 exerted anti-inflammatory effects and protected against acute cerebral ischemia/reperfusion injury possibly by acting through the IκB/NF-κB cascade to suppress microglia-mediated neuroinflammation.
（5R）-5-羟基雷公藤内酯醇（LLDT-8）是雷公藤内酯醇的类似物，与雷公藤内酯醇相比毒性更低，且具有相当的免疫抑制作用。我们研究了LLDT-8对脑缺血/再灌注损伤的抗炎和神经保护作用。
通过用格里斯试剂测量培养基中亚硝酸盐浓度来评估小胶质细胞产生一氧化氮的情况。通过qPCR和蛋白质印迹法检测小胶质细胞和缺血脑组织中促炎介质的表达。用TTC组织学评估梗死体积。用蛋白质印迹法和免疫细胞化学分析法分析TLR4/NF-κB信号通路。
LLDT-8显著减小了缺血皮层的梗死面积，并降低了促炎细胞因子的表达。LLDT-8抑制了经脂多糖处理的BV-2小胶质细胞和原代小胶质细胞中一氧化氮的释放以及肿瘤坏死因子-α、白细胞介素-1β和诱导型一氧化氮合酶的表达。此外，LLDT-8抑制了TLR4的表达、IκBα的降解以及NF-κB的核转位。
LLDT-8发挥了抗炎作用，并可能通过IκB/NF-κB级联反应抑制小胶质细胞介导的神经炎症，从而保护免受急性脑缺血/再灌注损伤。
