针刺研究2025，Vol.50Issue(7)：754-762,9.DOI:10.13702/j.1000-0607.20240274
针刺抑制铜死亡延长脑梗死大鼠溶栓时间窗的实验研究
Acupuncture inhibits cuproptosis to prolong the time window of thrombolysis by down-regulating cerebral FDX1 and DLAT in rats with cerebral infarction
摘要
Abstract
Objective To observe the effect of acupuncture on neuronal cuproptosis after thrombolytic treatment in rats with cerebral infarction(CI),so as to explore its underlying mechanisms in prolonging the time window of thrombolysis therapy for CI.Methods In the first part of the present study,40 male SD rats were randomly divided into 5 groups:sham operation,model,4.5 h thrombolysis,6 h thrombolysis and acupuncture+6 h thrombolysis groups,with 8 rats in each group.The CI model was prepared by modified autologous thromboembolism method.Rats of the 4.5 h thrombolysis,6 h thrombolysis and acupuncture+6 h thrombolysis groups received thrombolysis with intravenous injection of recombinant human tissue plasminogen activator(rt-PA,10 mg/kg)through tail vein at 4.5 h or 6 h after modeling.Manual acupuncture stimulation was applied to bilateral"Neiguan"(PC6)and"Shuigou"(CV26)at the same time with thrombolysis administration for rats of the acupuncture+6 h thrombolysis group,with the needles retained for 30 min.The cerebral blood flow was monitored using laser diffusion blood flow imaging,and neurological deficit score was evaluated 2 h and 24 h after successful modeling using Bederson's method.The CI volume was observed after triphenyltetrazole chloride staining.The immunoactivity of neuronal nuclei(NeuN)in the ischemic penumbra(IP)region of the cerebral cortex was detected by immunofluorescence staining.In the second part of the study,24 male SD rats were randomly divided into sham operation group,model group,6 h thrombolysis group and acupuncture+6 h thrombolysis group,with 6 rats in each group.The protein and mRNA relative expression levels of Ferredoxin-1(FDX1)and dihydrolipoamide acetyltransferase(DLAT)in the IP region of the right cerebral cortex were detected by Western blot and real-time PCR,separately.The copper ion,pyruvate and adenosine triphosphate(ATP)contents in the IP area of the cerebral cortex tissue were detected using colorimetry.Results The neurological deficit scores at both 2 h and 24 h after modeling,percentage of CI volume,the contents of copper ion and pyruvate,and the protein and mRNA expression levels of FDX1 and DLAT were significantly higher in the model group than those in the sham operation group(P<0.01),while the NeuN positive expression and ATP content were evidently lower in the model group than those in the sham operation group(P<0.01).In comparison with the model group,the neurological deficit score and percentage of CI volume in the 4.5 h thrombolysis and acupuncture+6 h thrombolysis groups,the contents of copper ion and pyruvate,and the protein and mRNA expression levels of FDX1 and DLAT in the acupuncture+6 h thrombolysis group,and the NeuN immunoactivity and ATP content in the 6 h thrombolysis group were significantly decreased(P<0.05,P<0.01),while the immunoactivity of NeuN in both 4.5 h thrombolysis and acupuncture+6 h thrombolysis groups,ATP content in the acupuncture+6 h thrombolysis group,the percentage of CI volume,contents of copper ion and pyruvate,and the expression levels of FDX1 and DLAT proteins and mRNAs in the 6 h thrombolysis group were significantly increased(P<0.05,P<0.01).Comparison among the 3 intervention groups showed that the neurological deficit score and percentage of CI volume were considerably lower in both of the 4.5 h thrombolysis and acupuncture+6 h thrombolysis groups than those in the 6 h thrombolysis group(P<0.01),while the NeuN immunoactivity was considerably higher in both of the 4.5 h thrombolysis and acupuncture+6 h thrombolysis groups than in the 6 h thrombolysis group(P<0.01).The contents of copper ion and pyruvate,and the protein and mRNA expression levels of FDX1 and DLAT were considerably lower in the acupuncture+6 h thrombolysis group than in the 6 h thrombolysis group(P<0.01),while the ATP content was considerably higher in the acupuncture+6 h thrombolysis group than in the 6 h thrombolysis group(P<0.01).Conclusion Acupuncture can prolong the time window of thrombolysis therapy for CI rats,which may be related to its function in down-regulating the protein and mRNA expressions of FDX1 and DLAT in ischemic cerebral cortex.关键词
脑梗死/针刺/溶栓时间窗/铜死亡Key words
Cerebral infarction/Acupuncture/Thrombolysis time window/Cuproptosis引用本文复制引用
赵艺,宋扬扬,常思琦,黄正,刘欣雨,刘倩倩,倪光夏..针刺抑制铜死亡延长脑梗死大鼠溶栓时间窗的实验研究[J].针刺研究,2025,50(7):754-762,9.基金项目
国家自然科学基金项目(No.82274639、82074525、82104993)) （No.82274639、82074525、82104993）
江苏省中医药科技发展计划项目(No.QN202204) （No.QN202204）
