Department of Acupuncture and Rehabilitation, Affiliated Hospital of Nanjing University of Chinese Medicine, Nanjing, 210029, Jiangsu Province, China.
Curr Neurovasc Res. 2024;21(1):64-73. doi: 10.2174/0115672026294183240207115956.
Electroacupuncture (EA) treatment plays a protective role in cerebral ischemiareperfusion (CIR) injury. However, the underlying molecular mechanism is still not fully elucidated.
All rats were randomly divided into five groups: the SHAM group, MCAO group, MCAO+EA (MEA) group, MCAO+METTL3 overexpression+EA (METTL3) group and MCAO+lncRNA H19 overexpression+EA (lncRNA H19) group. The middle cerebral artery occlusion (MCAO) rats were established to mimic CIR injury. The overexpression of lncRNA H19 and METTL3 was induced by stereotactic injection of lentiviruses into the rat lateral ventricles. The rats in the MEA, METTL3, and lncRNA H19 groups were treated with EA therapy on "Renzhong" (DU26) and "Baihui" (DU20) acupoints (3.85/6.25Hz; 1mA). Besides, the neurological deficit scoring, cerebral infarction area, pathological changes in brain tissue, total RNA m6A level, and the expression of METTL3, S1PR2, TLR4, NLRP3 and lncRNA H19 were detected in this experiment.
EA improved the neurological deficit scoring, cerebral infarction area, and pathological injury in MCAO rats, while these beneficial effects of EA on CIR injury were attenuated by the overexpression of METTL3 or lncRNA H19. More importantly, EA down-regulated the total RNA m6A level and the expression of METTL3, S1PR2, TLR4, NLRP3 and lncRNA H19 in MCAO rats. Instead, the overexpression of METTL3 or lncRNA H19 was found to reverse the EA-induced down-regulation.
The findings indicated that EA might down-regulate the S1PR2/TLR4/NLRP3 signaling pathway via m6A methylation of lncRNA H19 to alleviate CIR injury. Our findings provide a new insight into the molecular mechanism of EA on CIR injury.
电针（EA）治疗在脑缺血再灌注（CIR）损伤中发挥保护作用。然而，其潜在的分子机制仍未完全阐明。
所有大鼠随机分为五组：假手术（SHAM）组、大脑中动脉闭塞（MCAO）组、MCAO+EA（MEA）组、MCAO+METTL3 过表达+EA（METTL3）组和 MCAO+lncRNA H19 过表达+EA（lncRNA H19）组。通过立体定向注射慢病毒到大鼠侧脑室建立 CIR 损伤的 MCAO 大鼠模型。通过立体定向注射慢病毒到大鼠侧脑室诱导 lncRNA H19 和 METTL3 的过表达。MEA、METTL3 和 lncRNA H19 组大鼠在“人中”（DU26）和“百会”（DU20）穴位进行 EA 治疗（3.85/6.25Hz；1mA）。此外，本实验还检测了神经功能缺损评分、脑梗死面积、脑组织病理变化、总 RNA m6A 水平以及 METTL3、S1PR2、TLR4、NLRP3 和 lncRNA H19 的表达。
EA 改善了 MCAO 大鼠的神经功能缺损评分、脑梗死面积和病理损伤，而过表达 METTL3 或 lncRNA H19 则减弱了 EA 对 CIR 损伤的有益作用。更重要的是，EA 下调了 MCAO 大鼠的总 RNA m6A 水平和 METTL3、S1PR2、TLR4、NLRP3 和 lncRNA H19 的表达。相反，过表达 METTL3 或 lncRNA H19 则发现可逆转 EA 诱导的下调。
这些发现表明，EA 可能通过 lncRNA H19 的 m6A 甲基化下调 S1PR2/TLR4/NLRP3 信号通路，从而减轻 CIR 损伤。我们的研究结果为 EA 治疗 CIR 损伤的分子机制提供了新的见解。
