姜黄素通过激活AMPK信号通路介导的自噬抑制大鼠蛛网膜下腔出血后神经元凋亡  

Curcumin inhibits neuronal apoptosis following subarachnoid hemorrhage in rats by activating AMPK signaling mediated autophagy

作　　者：罗焱[1] 张平(指导)[1] LUO Yan;ZHANG Ping(Internal Medicine-Neurology,Affiliated Nanhua Hospital,University of South China,Heng-yang 421002,China)

机构地区：[1]南华大学附属南华医院神经内科,衡阳421002

出　　处：《中国免疫学杂志》2021年第7期807-811,818,共6页Chinese Journal of Immunology

基　　金：国家自然科学基金项目(81471310)。

摘　　要：目的:探讨姜黄素(Cur)对大鼠蛛网膜下腔出血(SAH)后神经元凋亡的影响及其作用机制。方法:将60只雄性SD大鼠随机分成5组:假手术组(Sham)、模型组(SAH)、Cur低剂量干预组(50 mg/kg,Cur-L)、Cur高剂量干预组(200 mg/kg,Cur-H)和高剂量Cur联合AMPK抑制剂Dorsomorphin干预组(200 mg/kg Cur+25 mg/kg Dorsomorphin,Cur-H+Dor),每组12只。除Sham组外,其余各组均采用血管内穿刺法建立大鼠SAH模型,术后立即给药,1次/d,连续3 d。评估各组大鼠神经功能情况并检测脑组织水肿程度;TUNEL染色法检测大鼠海马组织神经元凋亡情况;免疫荧光染色观察大鼠海马区LC3-Ⅱ蛋白颗粒形成情况;Western blot检测大鼠海马组织Beclin-1、LC3-Ⅱ、cleaved-caspase-3、AMPK、p-AMPK、mTOR和p-mTOR等蛋白表达水平。结果:与Sham组比较,SAH组大鼠脑组织含水量和海马组织神经元凋亡率均显著增加,神经功能评分和LC3-Ⅱ蛋白颗粒形成显著降低(P<0.05),且海马组织中cleaved-caspase-3和p-mTOR蛋白表达水平显著升高(P<0.05),而LC3-Ⅱ、Beclin-1、p-AMPK等蛋白表达水平显著降低(P<0.05);与SAH组相比,Cur-H组大鼠脑组织含水量以及海马神经细胞凋亡率均显著降低(P<0.05),神经功能评分和LC3-Ⅱ蛋白颗粒形成显著增加(P<0.05),同时海马组织中LC3-Ⅱ、Beclin-1、p-AMPK等蛋白表达水平均显著增加(P<0.05),而cleaved-caspase-3和p-mTOR蛋白表达水平显著降低(P<0.05),采用Dor联合干预可显著抑制高剂量Cur作用效果。结论:Cur能够改善大鼠SAH后神经损伤,其机制可能是通过激活AMPK信号通路介导的自噬进而抑制海马神经元凋亡。Objective:To investigate the effects of curcumin(Cur)on the neuronal apoptosis following subarachnoid hemorrhage(SAH)in rats and its mechanism.Methods:Sixty male SD rats were randomly divided into sham operation group(Sham),model group(SAH),low-dose Cur intervention group(50 mg/kg,Cur-L),high-dose Cur intervention group(200 mg/kg,Cur-H)and high dose Cur combined with AMPK inhibitor Dorsomorphin intervention group(200 mg/kg Cur+25 mg/kg Dorsomorphin,CurH+Dor),with 12 rats in each group.Except for sham group,the SAH rat models in the other groups were established by intravascular puncture.Then rats were administered with Cur or Dorsomorphin,once a day for 3 days.The neurological function and the brain water content of rats in each group were evaluated.TUNEL staining was used to detect neuronal apoptosis in rat hippocampal tissue.Immunofluorescence staining was used to observe the formation of LC3-Ⅱin the hippocampus of rats.Western blot analysis was used to detect the protein expression levels of Beclin-1,LC3-Ⅱ,cleaved-caspase-3,AMPK,p-AMPK,mTOR and p-mTOR in the hippocampus of rats.Results:Compared with Sham group,the water content of brain tissue and the apoptosis rate of hippocampal neurons in the SAH group were significantly increased(P<0.05),the neurological function score and the formation of LC3-Ⅱprotein granules were significantly decreased(P<0.05),and the protein expression levels of cleaved-caspase-3 and p-mTOR in the hippocampus were significantly increased(P<0.05),while the protein expression levels of LC3-Ⅱ,Beclin-1 and p-AMPK were significantly decreased(P<0.05).Compared with SAH group,the water content of brain tissue and the apoptosis rate of hippocampal neurons in Cur-H group were significantly decreased(P<0.05),and the neurological function score and the formation of LC3-Ⅱprotein granules were significantly increased(P<0.05).The protein expression levels of LC3-Ⅱ,Beclin-1 and p-AMPK in hippocampus were significantly increased(P<0.05),while the protein expression levels of cleaved-caspa

关 键 词：姜黄素 AMPK信号通路 神经元 自噬 

分 类 号：R743.35[医药卫生—神经病学与精神病学]