Liu Fang, Chen Ying, Huang Kangbai
Department of Chinese Medicine, The First Affiliated Hospital of Guangdong Pharmaceutical University, Guangzhou, China.
Medical College of Acu-Moxi and Rehabilitation, Guangzhou University of Chinese Medicine, No. 12, Jichang Road, Baiyun District, Guangzhou, 510405, China.
Appl Biochem Biotechnol. 2025 Feb;197(2):989-1001. doi: 10.1007/s12010-024-05063-6. Epub 2024 Sep 28.
Electro-acupuncture (EA) is identified as an effective therapeutic method for cerebral ischemia/reperfusion injury (CIRI), which is a combination of Chinese traditional acupuncture and modern electro-therapy. However, the downstream molecular mechanisms of EA in CIRI process remains largely unknown. The purpose of the present study is to unveil the therapeutic effect of EA on CIRI rat and its regulatory mechanisms. At first, we constructed middle cerebral artery occlusion (MCAO) rat models and then treated them with EA to observe the pathological changes. The results indicated that EA decreased the infarct volume (43.81 ± 3.34 vs 15.96 ± 2.22) and the neurological scores (3.33 ± 0.52 vs 1.67 ± 0.52) and suppressed the apoptosis in MCAO model rats. For ferroptosis analysis, EA decreased the Fe2 + (0.08 ± 0.01 vs 0.06 ± 0.01), MDA (36.61 ± 4.29 vs 21.72 ± 2.79), and LPS (5.25 ± 0.69 vs 2.89 ± 0.42) contents and increased the GSH (4.94 ± 1.04 vs 11.69 ± 1.88) content in MCAO model rats. We next detected whether succinylation mediated EA-treated I/R injury. According to immunoprecipitation and western blot analysis, EA treatment could lower both levels of succinylation and KAT3B in MCAO rats. Moreover, mechanism experiments unveiled that KAT3B promoted the succinylation of the ferroptosis-related protein ACSL4 at K661 site and thus stabilizing ACSL4. Finally, EA-treated MCAO rats were further injected with KAT3B expression vector. The results showed that KAT3B overexpression increased the infarct volume (31.44 ± 3.92 vs 7.94 ± 2.84) and the neurological scores (2.67 ± 0.51 vs 1.33 ± 0.51) and promoted the apoptosis in EA treated MCAO model rats. For ferroptosis analysis, KAT3B overexpression increased the Fe2 + (0.08 ± 0.01 vs 0.05 ± 0.01), MDA (29.24 ± 4.30 vs 22.06 ± 1.89), and LPO (5.07 ± 0.45 vs 2.88 ± 0.49) contents and decreased the GSH (7.86 ± 1.09 vs 11.06 ± 1.76) content in EA treated MCAO model rats. Collectively, our study demonstrates that EA plays a therapeutic role in CIRI through suppressing KAT3B-induced stabilization of ACSL4 to inhibit ferroptosis. These findings contribute to our understanding of the molecular mechanisms underlying the neuroprotective effects of EA and open new avenues for the development of innovative therapeutic strategies for CIRI.
电针（EA）被认为是治疗脑缺血/再灌注损伤（CIRI）的一种有效治疗方法，它是传统中医针灸与现代电疗法的结合。然而，EA在CIRI过程中的下游分子机制仍 largely 未知。本研究的目的是揭示EA对CIRI大鼠的治疗作用及其调控机制。首先，我们构建了大脑中动脉闭塞（MCAO）大鼠模型，然后用EA对其进行治疗以观察病理变化。结果表明，EA降低了梗死体积（43.81±3.34 vs 15.96±2.22）和神经学评分（3.33±0.52 vs 1.67±0.52），并抑制了MCAO模型大鼠的细胞凋亡。对于铁死亡分析，EA降低了MCAO模型大鼠中Fe2+（0.08±0.01 vs 0.06±0.01）、丙二醛（MDA，36.61±4.29 vs 21.72±2.79）和脂多糖（LPS，5.25±0.69 vs 2.89±0.42）的含量，并增加了谷胱甘肽（GSH，4.94±1.04 vs 11.69±1.88）的含量。接下来，我们检测琥珀酰化是否介导了EA治疗的I/R损伤。根据免疫沉淀和蛋白质印迹分析，EA治疗可降低MCAO大鼠的琥珀酰化水平和KAT3B水平。此外，机制实验表明，KAT3B促进铁死亡相关蛋白ACSL4在K661位点的琥珀酰化，从而稳定ACSL4。最后，对EA治疗的MCAO大鼠进一步注射KAT3B表达载体。结果表明，KAT3B过表达增加了梗死体积（31.44±3.92 vs 7.94±2.84）和神经学评分（2.67±0.51 vs 1.33±0.51），并促进了EA治疗的MCAO模型大鼠的细胞凋亡。对于铁死亡分析，KAT3B过表达增加了EA治疗的MCAO模型大鼠中Fe2+（0.08±0.01 vs 0.05±0.01）、MDA（29.24±4.30 vs 22.06±1.89）和脂质过氧化物（LPO，5.07±0.45 vs 2.
