安五脂素对氧化应激诱导的海马神经元HT22细胞自噬和凋亡的影响  

Effect of Macelignan on the autophagy and apoptosis of hippocampal neuron HT22 cells induced by oxidative stress

作　　者：李莎 楚新歌 邱新茹 李莉 延光海[1,2] 崔春爱 LI Sha;CHU Xin-ge;QIU Xin-ru;LI Li;YAN Cuang-hai;CUI Chun-ai(Department of Anatomy,Medical College of Yanbian University,Yanji 133000,Jilin Province,China;Jilin Key Laboratory for Immune and Targeting Research on Common Allergic Diseases,Medical College of Yanbian University,Yanji 133000,Jilin Province,China)

机构地区：[1]延边大学医学院解剖学教研室,吉林延吉133000 [2]延边大学医学院吉林省科技厅过敏性常见疾病免疫与靶向研究重点实验室,吉林延吉133000

出　　处：《中国临床药理学杂志》2024年第13期1865-1868,共4页The Chinese Journal of Clinical Pharmacology

基　　金：国家自然科学基金资助项目(82060746;81660687)。

摘　　要：目的探讨安五脂素对氧化应激介导的神经元自噬和凋亡的干预作用。方法选用鼠性海马神经元细胞,用2.5 mmol·L^(-1)谷氨酸(Glu)制备氧化应激细胞模型。将细胞分为正常组(正常培养)、模型组(2.5 mmol·L^(-1)Glu)和低、中、高剂量实验组(2.5、5、10μmol·L^(-1)安五脂素处理)、抑制药组(2.5 mmol·L^(-1)Glu+10μmol·L^(-1)安五脂素+10μmol·L^(-1)LY294002)。用流式细胞术检测细胞凋亡率;用蛋白印迹法检测自噬微管相关蛋白轻链3蛋白(LC3B)、泛素结合蛋白(p62)、细胞周期蛋白p21、B淋巴细胞瘤-2(Bcl-2)和Bcl-2相关X蛋白(Bax)表达水平。结果正常组、模型组和低、中、高剂量实验组的凋亡率分别为(4.58±1.25)%、(8.75±0.55)%、(6.30±1.71)%、(5.97±2.27)%和(5.49±1.71)%。模型组与正常组比较,在统计学上差异有统计学意义(P<0.01);低、中、高剂量实验组与模型组比较,在统计学上差异均有统计学意义(均P<0.01)。正常组、模型组和低、中、高剂量实验组及抑制药组LC3B蛋白水平分别为0.28±0.02、0.74±0.02、1.02±0.04、0.70±0.03、0.26±0.02和0.21±0.01;p62蛋白水平分别为0.49±0.08、0.33±0.03、0.50±0.07、0.59±0.01、0.64±0.13和0.65±0.06;p21蛋白水平分别为0.87±0.02、1.18±0.03、0.98±0.03、0.88±0.03、0.72±0.06和0.81±0.02;Bcl-2/Bax蛋白水平分别为1.74±0.23、1.11±0.10、1.38±0.05、1.66±0.26、1.58±0.29和1.53±0.09。上述指标,模型组与正常组比较,高剂量实验组与模型组比较,抑制药组与模型组比较,在统计学上差异均有统计学意义(均P<0.01)。结论安五脂素可通过调控细胞自噬和凋亡过程,减轻谷氨酸所诱导的海马神经元损伤,有明显的神经保护作用。Objective To explore the regulatory mechanism of Macelignan on oxidative stress-mediated neuronal injury in autophagy and apoptosis.Methods Murine hippocampal neuronal HT22 cells were treated with 2.5 mmol·L^(-1)glutamic acid(Glu)to establish an oxidative stress cell model.The cells were divided into normal group(normal cultured cells),model group(2.5 mmol·L^(-1)Glu)and experimental-L,-M,-H groups(2.5,5,10μmol·L^(-1)Macelignan treatment),inhibitor group(2.5 mmol·L^(-1)Glu+10μmol·L^(-1)Macelignan+10μmol·L^(-1)LY294002).Aoptosis rate was detected by flow cytometry;the protein expression level of autophagy-related protein LC3B(LC3B),anti-SQSTM1/p62(p62),p21,B-cell lymphoma-2(Bcl-2)and Bel-2 associated X protein(Bax)was detected by Western blot.Results The apoptosis rates in the normal group,model group and experimental-L,-M,-H groups were(4.58±1.25)%,(8.75±0.55)%,(6.30±1.71)%,(5.97±2.27)%and(5.49±1.71)%.The difference between model group and normal group was statistically significant(P<0.01).The difference between experimental-L,-M,-H groups and model group was statistically significant(all P<0.01).The levels of LC3B in normal group,model group,experimental-L,experimental-M,experimental-H groups and inhibitor group were 0.28±0.02,0.74±0.02,1.02±0.04,0.70±0.03,0.26±0.02 and 0.21±0.01;p62 levels were 0.49±0.08,0.33±0.03,0.50±0.07,0.59±0.01,0.64±0.13 and 0.65±0.06;p21 levels were 0.87±0.02,1.18±0.03,0.98±0.03,0.88±0.03,0.72±0.06and 0.81±0.02;Bcl-2/Bax levels were 1.74±0.23,1.11±0.10,1.38±0.05,1.66±0.26,1.58±0.29 and1.53±0.09,respectively.The differences between model group and normal group,between model group and experimental-H group,between model group and inhibitor group,were also statistically significant(all P<0.01).Conclusion Macelignan can reduce the damage of hippocampal neurons induced by glutamate acid by regulating the process of autophagy and apoptosis,and has obvious neuroprotective effect.

关 键 词：安五脂素 神经元 氧化应激 自噬 凋亡 

分 类 号：R28[医药卫生—中药学]