College of Life and Health Sciences, Northeastern University, Shenyang, China; Key Laboratory of Data Analytics and Optimization for Smart Industry, Northeastern University, Ministry of Education, Shenyang, China.
School of Chemical Science and Technology, Yunnan University, Kunming, China.
J Ethnopharmacol. 2021 Apr 6;269:113718. doi: 10.1016/j.jep.2020.113718. Epub 2020 Dec 25.
Ferula sinkiangensis K. M. Shen is a traditional Chinese medicine that has a variety of pharmacological properties relevant to neurological disorders and inflammations. Kellerin, a novel compound extracted from Ferula sinkiangensis, exerts a strong anti-neuroinflammatory effect by inhibiting microglial activation. Microglial activation plays a vital role in ischemia-induced brain injury. However, the potential therapeutic effect of kellerin on focal cerebral ischemia is still unknown.
To explore the effect of kellerin on cerebral ischemia and clarify its possible mechanisms, we applied the middle cerebral artery occlusion (MCAO) model and the LPS-activated microglia model in our study.
Neurological outcome was examined according to a 4-tiered grading system. Brain infarct size was measured using TTC staining. Brain edema was calculated using the wet weight minus dry weight method. Neuron damage and microglial activation were observed by immunofluorescence in MCAO model in rats. In in vitro studies, microglial activation was examined by flow cytometry and the viability of neuronal cells cultured in microglia-conditioned medium was measured using MTT assay. The levels of pro-inflammatory cytokines were measured by qRT-PCR and ELISA. The proteins involved in NF-κB signaling pathway were determined by western blot. Intracellular ROS was examined using DCFH-DA method and NADPH oxidase activity was measured using the NBT assay.
We found that kellerin improved neurological outcome, reduced brain infarct size and decreased brain edema in MCAO model in rats. Under the pathologic conditions of focal cerebral ischemia, kellerin alleviated neuron damage and inhibited microglial activation. Moreover, in in vitro studies of LPS-stimulated BV2 cells kellerin protected neuronal cells from being damaged by inhibiting microglial activation. Kellerin also reduced the levels of pro-inflammatory cytokines, suppressed the NF-κB signaling pathway, and decreased ROS generation and NADPH oxidase activity.
Our discoveries reveal that the neuroprotective effects of kellerin may largely depend on its inhibitory effect on microglial activation. This suggests that kellerin could serve as a novel anti-inflammatory agent which may have therapeutic effects in ischemic stroke.
新疆阿魏（Ferula sinkiangensis K. M. Shen）是一种传统中药，具有多种与神经紊乱和炎症相关的药理学特性。从新疆阿魏中提取的新型化合物凯勒林（kellerin）通过抑制小胶质细胞激活发挥强大的抗神经炎症作用。小胶质细胞激活在缺血性脑损伤中起着至关重要的作用。然而，凯勒林对局灶性脑缺血的潜在治疗效果尚不清楚。
通过应用大脑中动脉闭塞（MCAO）模型和 LPS 激活的小胶质细胞模型，探讨凯勒林对脑缺血的影响，并阐明其可能的机制。
根据四级分级系统评估神经功能结果。采用 TTC 染色测量脑梗死面积。采用湿重-干重法计算脑水肿。通过免疫荧光观察 MCAO 模型中大鼠的神经元损伤和小胶质细胞激活。在体外研究中，通过流式细胞术检测小胶质细胞激活，并用 MTT 测定法测量培养在小胶质细胞条件培养基中的神经元细胞活力。通过 qRT-PCR 和 ELISA 测定促炎细胞因子水平。通过 Western blot 测定 NF-κB 信号通路相关蛋白。通过 DCFH-DA 法检测细胞内 ROS，通过 NBT 测定法测量 NADPH 氧化酶活性。
我们发现凯勒林可改善 MCAO 模型中大鼠的神经功能，减少脑梗死面积和脑水肿。在局灶性脑缺血的病理条件下，凯勒林减轻了神经元损伤并抑制了小胶质细胞激活。此外，在 LPS 刺激的 BV2 细胞的体外研究中，凯勒林通过抑制小胶质细胞激活来保护神经元细胞免受损伤。凯勒林还降低了促炎细胞因子的水平，抑制了 NF-κB 信号通路，并减少了 ROS 生成和 NADPH 氧化酶活性。
我们的发现表明，凯勒林的神经保护作用可能在很大程度上取决于其对小胶质细胞激活的抑制作用。这表明凯勒林可能作为一种新型抗炎药物，在缺血性中风中具有治疗作用。
