Department of Neurology and Neurosurgery & Clinical College of Neurorehabilitation of Tianjin Medical University, Tianjin, 300350, China; Department of Neurology, Affiliated Hospital of Hebei University, Baoding, 071030, China.
Department of Neurology, Tianjin Huanhu Hospital, Tianjin, 300350, China.
Neurotoxicology. 2021 Dec;87:62-69. doi: 10.1016/j.neuro.2021.08.018. Epub 2021 Sep 2.
Parkinson's disease (PD) is a complicated multifactorial neurodegenerative disorder. Oxidative stress, neuroinflammatory response, and activation of apoptosis have been proposed to be tightly involved in the pathogenesis of PD. Genkwanin is a typical bioactive non-glycosylated flavonoid with anti-inflammatory and anti-oxidant activities. However, the effect of genkwanin on PD remains unclear. Cell viability, lactate dehydrogenase (LDH) release, caspase-3/7 activity, and apoptosis was evaluated by MTT, LDH release assay, caspase-3/7 activity assay, and TUNEL assay, respectively. The secretion of prostaglandin E (PGE), tumor necrosis factor (TNF)-α, interleukin (IL)-1β, and IL-6 were measured by respective commercial ELISA kits. The mRNA expression of TNF-α, IL-1β, and IL-6 was detected by qRT-PCR. The protein levels of cycloxygenase-2 (COX-2), toll-like receptor 4 (TLR4), myeloid differentiation factor 88 (MyD88), and NOD-like receptor (NLR) protein: 3 (NLRP3) were determined by western blot analysis. Genkwanin at concentrations less than 40 μM had no impact on cell viability and LDH release. Genkwanin suppressed MPP-induced neuroinflammation in SH-SY5Y cells. MPP treatment inhibited cell viability, increased LDH release, apoptosis, and ROS generation, and reduced superoxide dismutase (SOD) activity in SH-SY5Y cells, which were abolished by genkwanin treatment. Genkwanin suppressed MPP-induced activation of TLR4/MyD88/NLRP3 inflammasome pathway in SH-SY5Y cells. TLR4 overexpression weakened the anti-inflammatory and anti-neurotoxicity of genkwanin in SH-SY5Y cells. In conclusion, genkwanin attenuated neuroinflammation and neurotoxicity by inhibiting TLR4/MyD88/NLRP3 inflammasome pathway in MPP-induced cellular model of PD.
帕金森病 (PD) 是一种复杂的多因素神经退行性疾病。氧化应激、神经炎症反应和细胞凋亡的激活被认为与 PD 的发病机制密切相关。根皮苷是一种具有抗炎和抗氧化活性的典型生物活性非糖基化类黄酮。然而，根皮苷对 PD 的影响尚不清楚。通过 MTT、LDH 释放测定、caspase-3/7 活性测定和 TUNEL 测定分别评估细胞活力、乳酸脱氢酶 (LDH) 释放、caspase-3/7 活性和细胞凋亡。通过各自的商业 ELISA 试剂盒测量前列腺素 E (PGE)、肿瘤坏死因子 (TNF)-α、白细胞介素 (IL)-1β 和 IL-6 的分泌。通过 qRT-PCR 检测 TNF-α、IL-1β 和 IL-6 的 mRNA 表达。通过 Western blot 分析测定环加氧酶-2 (COX-2)、Toll 样受体 4 (TLR4)、髓样分化因子 88 (MyD88) 和 NOD 样受体 (NLR) 蛋白 3 (NLRP3) 的蛋白水平。浓度低于 40 μM 的根皮苷对细胞活力和 LDH 释放没有影响。根皮苷抑制 MPP 诱导的 SH-SY5Y 细胞神经炎症。MPP 处理抑制细胞活力，增加 LDH 释放、凋亡和 ROS 生成，并降低 SH-SY5Y 细胞中超氧化物歧化酶 (SOD) 活性，根皮苷处理可消除这些作用。根皮苷抑制 MPP 诱导的 SH-SY5Y 细胞 TLR4/MyD88/NLRP3 炎性小体通路的激活。TLR4 过表达减弱了根皮苷在 SH-SY5Y 细胞中的抗炎和神经保护作用。总之，根皮苷通过抑制 TLR4/MyD88/NLRP3 炎性小体通路减轻 MPP 诱导的 PD 细胞模型中的神经炎症和神经毒性。
