Title:Electroacupuncture Alleviates Cerebral Ischemia-reperfusion Injury by
Regulating the S1PR2/TLR4/NLRP3 Signaling Pathway via m6A Methylation
of lncRNA H19
Volume: 21
Issue: 1
Author(s): Han-Rui Zhang, Gu-Quan Ma, He-Qun Lv, Yao-Ting Feng and Yong-Jun Peng*
Affiliation:
- Department of Acupuncture and Rehabilitation, Affiliated Hospital of Nanjing University of Chinese Medicine,
Nanjing, 210029, Jiangsu Province, China
Keywords:
Electroacupuncture, cerebral ischemia-reperfusion, S1PR2/TLR4/NLRP3, m6A methylation, lncRNA H19, METTL3.
Abstract:
Electroacupuncture (EA) treatment plays a protective role in cerebral ischemiareperfusion
(CIR) injury. However, the underlying molecular mechanism is still not fully elucidated.
Methods: 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.
Results: 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.
Conclusion: 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.