Zhang Heng-Sheng, Ouyang Bo, Ji Xiong-Ying, Liu Mei-Fang
Department of Rehabilitation, Affiliated Nanhua Hospital, University of South China, Hengyang, 421002, Hunan, People's Republic of China.
Department of Traditional Chinese Medicine, Affiliated Nanhua Hospital, University of South China, No. 336 Dongfeng South Road, Zhuhui District, Hengyang, 421002, Hunan, People's Republic of China.
Neurochem Res. 2021 Jul;46(7):1747-1758. doi: 10.1007/s11064-021-03285-2. Epub 2021 Apr 11.
Cerebral ischaemia/reperfusion (I/R) injury-induced irreversible brain injury is a major cause of mortality and functional impairment in ageing people. Gastrodin (GAS), derived from the traditional Chinese herbal medicine Tianma, has been reported to inhibit the progression of stroke, but the mechanism whereby GAS modulates the progression of cerebral I/R remains unclear. The middle cerebral artery occlusion method was used as a model of I/R in vivo. Rats were pretreated with GAS by intraperitoneal injection 7 days before I/R surgery and were then treated with GAS for 7 days after I/R surgery. Additionally, an oxygen-glucose deprivation/reoxygenation model using neuronal cells was established in vitro to simulate I/R injury. 2,3,5-Triphenyltetrazolium chloride and Nissl staining were used to evaluate infarct size and neuronal damage, respectively. Lactate dehydrogenase release and cell counting kit-8 assays were used to assess neuronal cell viability. Enzyme-linked immunosorbent assay, qPCR, flow cytometry and western blotting were performed to analyse the expression levels of inflammatory factors (IL-1β, IL-18), lncRNA NEAT1, miR-22-3p, NLRP3 and cleaved caspase-1. Luciferase reporter experiments were performed to verify the association between lncRNA NEAT1 and miR-22-3p. The results indicated that GAS could significantly improve the neurological scores of rats and reduce the area of cerebral infarction. Meanwhile, GAS inhibited pyroptosis by downregulating NLRP3, inflammatory factors (IL-1β, IL-18) and cleaved caspase-1. In addition, GAS attenuated I/R-induced inflammation in neuronal cells through the modulation of the lncRNA NEAT1/miR-22-3p axis. GAS significantly attenuated cerebral I/R injury via modulation of the lncRNA NEAT1/miR-22-3p axis. Thus, GAS might serve as a new agent for the treatment of cerebral I/R injury.
脑缺血/再灌注（I/R）损伤所致的不可逆脑损伤是老年人死亡和功能障碍的主要原因。天麻素（GAS）源自传统中药天麻，据报道可抑制中风进展，但GAS调节脑I/R进展的机制尚不清楚。采用大脑中动脉闭塞法建立体内I/R模型。在I/R手术前7天通过腹腔注射对大鼠进行GAS预处理，然后在I/R手术后用GAS治疗7天。此外，在体外建立了使用神经元细胞的氧糖剥夺/复氧模型以模拟I/R损伤。分别使用2,3,5-三苯基氯化四氮唑和尼氏染色评估梗死面积和神经元损伤。使用乳酸脱氢酶释放和细胞计数试剂盒-8检测评估神经元细胞活力。进行酶联免疫吸附测定、qPCR、流式细胞术和蛋白质印迹分析炎症因子（IL-1β、IL-18）、lncRNA NEAT1、miR-22-3p、NLRP3和裂解的半胱天冬酶-1的表达水平。进行荧光素酶报告基因实验以验证lncRNA NEAT1与miR-22-3p之间的关联。结果表明，GAS可显著改善大鼠神经功能评分并减少脑梗死面积。同时，GAS通过下调NLRP3、炎症因子（IL-1β、IL-18）和裂解的半胱天冬酶-1抑制细胞焦亡。此外，GAS通过调节lncRNA NEAT1/miR-22-3p轴减轻I/R诱导的神经元细胞炎症。GAS通过调节lncRNA NEAT1/miR-22-3p轴显著减轻脑I/R损伤。因此，GAS可能成为治疗脑I/R损伤的新型药物。
