针刺研究2023，Vol.48Issue(12)：1193-1201,9.DOI:10.13702/j.1000-0607.20230477
大鼠海马组织circHDAC2/miR-3065/SLC30A3轴参与针刺调节脑缺血再灌注损伤的初步研究
A preliminary investigation of the regulatory effect of acupuncture on hippocampal circHDAC2/miR-3065/SLC30A3 axis in CIRI rats
摘要
Abstract
Objective To explore the mechanisms of acupuncture against cerebral ischemia/reperfusion injury(CIRI)through observing the expression of circular RNA HDAC2(circHDAC2)in the hippocampus of rats.Methods SD rats were randomly divided into sham-operation,model and acupuncture groups,with 13 rats in each group.The rat model of CIRI was established by middle cerebral artery occlusion.In the acupuncture group,acupuncture was deliv-vered at"Dazhui"(GV14),"Shuigou"(GV26)and"Baihui"(GV20),and the needles were retained for 30 min each time and acupuncture was conducted once every 12 h for a total of 7 sessions.Before and after intervention,using modified Garcia scale,the neurological function of the rats were evaluated,and TTC staining was employed to deter-mine the cerebral infarct area.Gene chip technology was used to screen the circRNAs with differential expressions in the ischemic hippocampus,and the circRNAs with co-differential expression(co-DE circRNAs)in the model group/sham-operation group,and the acupuncture group/model group separately.Among those circRNAs,the core circRNAs were screened according to P value,fold change(FC)and gene ontology(GO)analysis;and their expressions in the ischemic hippocampus were determined using quantitative real-time PCR(qPCR).Based on the verification results,a competing endogenous RNA(ceRNA)prediction network was constructed.The expression levels of microRNA(miRNA)and mRNA with high node centrality in the prediction network were validated by qPCR.Results Before inter-vention,compared with the sham-operation group,the modified Garcia score of each modeling group decreased(P<0.01).After intervention,the modified Garcia score was reduced and the cerebral infarct area ratio increased(P<0.01)in the model group when compared with the sham-operation group.In the acupuncture group,the modified Garcia score was higher and the cerebral infarct area ratio lower(P<0.01)than those of the model group.The microarray results of gene chip showed that 16 co-DE circRNAs were down-regulated in the model group and up-regulated in the acupunc-ture group,and 7 co-DE circRNAs up-regulated in the model group and down-regulated in the acupuncture group.The core circHDAC2 and circNTRK2 were screened according to P value,FC and the enrichment number of GO entries.QPCR results showed that,compared with the sham-operation group,the expression of circHDAC2 and circNTRK2 of the ischemic hippocampal tissue was down-regulated in the model group(P<0.01);and that of circHDAC2 and circNTRK2 up-regulated in the acupuncture group when compared with the model group(P<0.01).The relevant ceRNA regulatory network was constructed for circHDAC2 and the prediction results showed that the regulatory networks con-tained 12 miRNAs and 31 mRNAs.Results of verifying miRNA with high network node centrality and mRNA relevant with nerve regulation showed that,when compared with the sham-operation group,the expression levels of miR-29a,miR-29b and the solute carrier family 30 member 3(SLC30A3)mRNA in the ischemic hippocampus were down-regulated(P<0.01);and those of miR-3065 and mercaptopyruvate sulfurtransferase(MPST)up-regulated(P<0.01)in the model group.Compared with the model group,the expressions of miR-29a,miR-29b and SLC30A3 mRNA in the ischemic hippocampus were up-regulated(P<0.01,P<0.05),while that of miR-3065 down-regulated(P<0.05)in the acupuncture group.Conclusion Acupuncture significantly improves the neurological function and reduces the cerebral infarct area in CIRI rats,which may be related to the regulation of hippocampal circHDAC2/miR-3065/SLC30A3 axis.关键词
脑缺血再灌注损伤/针刺/环状RNA/微小RNAKey words
Cerebral ischemia/reperfusion injury/Acupuncture/Circular RNA/MicroRNA分类
医药卫生引用本文复制引用
谢灿明,江姗姗,王瑶,向静,刘晓月,陈楚淘,田浩梅..大鼠海马组织circHDAC2/miR-3065/SLC30A3轴参与针刺调节脑缺血再灌注损伤的初步研究[J].针刺研究,2023,48(12):1193-1201,9.基金项目
国家自然科学基金项目(No.82274662) （No.82274662）
湖南省自然科学基金项目(No.2021JJ30490) （No.2021JJ30490）
长沙市科技局自然科学基金项目(No.kq2014094) （No.kq2014094）
湖南省研究生科研创新项目(No.2022CX103) （No.2022CX103）
