Mn-SOD Upregulation by Electroacupuncture Attenuates Ischemic Oxidative Damage via CB1R-Mediated STAT3 Phosphorylation
Mn-SOD Upregulation by Electroacupuncture Attenuates Ischemic Oxidative Damage via CB1R-Mediated STAT3 Phosphorylation
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DOI：
10.1007/s12035-014-8971-7
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发表时间：
2014-11
影响因子：
5.1
通讯作者：
Sisi Sun;Xiyao Chen;Yang Gao;Zhaoyu Liu;Qian Zhai;L. Xiong;M. Cai;Qiang Wang
中科院分区：
文献类型：
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作者：
Sisi Sun;Xiyao Chen;Yang Gao;Zhaoyu Liu;Qian Zhai;L. Xiong;M. Cai;Qiang Wang
电针(EA)预处理通过大麻素受体1型受体(CB1R)发挥抗脑缺血损伤的神经保护作用。本研究旨在通过CB1R研究信号转导和转录激活子3(STAT3)和锰超氧化物歧化酶(Mn-SOD)是否参与电针预适应的抗氧化作用。电针预处理后2h，C57BL/6小鼠局灶性脑缺血损伤模型为大脑中动脉阻断60min。用Western印迹和免疫荧光染色检测再灌注后2 h半影区Mn-SOD的表达。在有或无Mn-SOD小干扰RNA(SiRNA)存在或不存在的情况下，评价神经功能评分、脑梗塞体积、末端脱氧核苷酸转移酶介导的dUDP-生物素缺口末端标记(TUNEL)染色和氧化应激。此外，在有或没有CB1R拮抗剂(AM251，SR141716)和CB1R激动剂(花生四烯基-2-氯乙胺(ACEA)，Win 55,212-2)存在和不存在的情况下，还检测了Y705的Mn-SOD蛋白表达和STAT3的磷酸化。电针预处理可上调再灌流后2小时脑组织中锰超氧化物歧化酶蛋白的表达和锰超氧化物歧化酶阳性神经细胞的数量。电针预处理还可减轻氧化应激，抑制细胞凋亡，诱导缺血损伤的神经保护作用，而电针预处理的这些有益作用可被锰超氧化物歧化酶的下调所逆转。两种CB1R拮抗剂可阻断电针预处理引起的锰-超氧化物歧化酶表达上调和STAT3磷酸化，而两种CB1R激动剂可增加锰-超氧化物歧化酶的表达和STAT3的磷酸化水平。电针上调Mn-SOD可通过CB1R介导的STAT3磷酸化减轻卒中小鼠的缺血性氧化损伤，这可能是电针预处理诱导脑缺血保护的一种新机制。
Electroacupuncture (EA) pretreatment elicits the neuroprotective effect against cerebral ischemic injury through cannabinoid receptor type 1 receptor (CB1R). In current study, we aimed to investigate whether the signal transducer and activator of transcription 3 (STAT3) and manganese superoxide dismutase (Mn-SOD) were involved in the antioxidant effect of EA pretreatment through CB1R. At 2 h after EA pretreatment, focal cerebral ischemic injury was induced by transient middle cerebral artery occlusion for 60 min in C57BL/6 mice. The expression of Mn-SOD in the penumbra was assessed by Western blot and immunoflourescent staining at 2 h after reperfusion. In the presence or absence of Mn-SOD small interfering RNA (siRNA), the neurological deficit score, the infarct volume, the terminal deoxynucleotidyl transferase-mediated dUDP-biotin nick end labeling (TUNEL) staining, and oxidative stress were evaluated. Furthermore, the Mn-SOD protein expression and phosphorylation of STAT3 at Y705 were also determined in the presence and absence of CB1R antagonists (AM251, SR141716) and CB1R agonists (arachidonyl-2-chloroethylamide (ACEA), WIN 55,212-2). EA pretreatment upregulated the Mn-SOD protein expression and Mn-SOD-positive neuronal cells at 2 h after reperfusion. EA pretreatment also attenuated oxidative stress, inhibited cellular apoptosis, and induced neuroprotection against ischemic damage, whereas these beneficial effects of EA pretreatment were reversed by knockdown of Mn-SOD. Mn-SOD upregulation and STAT3 phosphorylation by EA pretreatment were abolished by two CB1R antagonists, while pretreatment with two CB1R agonists increased the expression of Mn-SOD and phosphorylation level of STAT3. Mn-SOD upregulation by EA attenuates ischemic oxidative damage through CB1R-mediated STAT3 phosphorylation in stroke mice, which may represent one new mechanism of EA pretreatment-induced neuroprotection against cerebral ischemia.