槲皮苷对Aβ25-35诱导的PC12细胞损伤的影响  

Effect of quercitrin on PC12 cell injury indued by Aβ25-35

作　　者：李树良[1] 王东[1] 李放 张建军[1] 马景鑑[3] Li Shuliang;Wang Dong;Li Fang;Zhang Jianjun;Ma Jingjian(Department of Neurosurgery,Tianjin Fourth Central Hospital,Tianjin 300140,China;Department of Neurosurgery,Peking University People’s Hospital,Beijing 100191,China;Department of Neurosurgery,Tianjin First Central Hospital,Tianjin 300192,China)

机构地区：[1]天津市第四中心医院神经外科,300140 [2]北京大学人民医院神经外科,100191 [3]天津市第一中心医院神经外科,300192

出　　处：《中华实验外科杂志》2020年第10期1851-1854,共4页Chinese Journal of Experimental Surgery

基　　金：天津市自然科学基金(17JCYBJC27300)。

摘　　要：目的:观察槲皮苷对β淀粉样蛋白25-35(Aβ25-35)诱导的PC12细胞损伤的影响。方法:将PC12细胞随机分为正常组,Aβ25-35组,槲皮苷低、中、高剂量组。噻唑蓝(MTT)法检测细胞活力,流式双染检测细胞凋亡,比色法检测含半胱氨酸的天冬氨酸蛋白水解酶3(Caspase-3)、Caspase-9活性,蛋白质印迹法(Western blot)检测各组细胞中B细胞淋巴瘤/白血病-2(bcl-2)、bcl-2相关X蛋白(bax)、磷脂酰肌醇-3-羟激酶/蛋白激酶B(PI3K/Akt)信号通路激活,组间比较采用t检验。结果:与正常组比较,Aβ25-35组细胞活力显著降低(0.362±0.036比0.453±0.045,t=4.230,P<0.05),细胞早期凋亡率[(17.89±1.81)%比(2.31±0.23)%,t=8.623,P<0.05]及晚期凋亡率[(19.56±1.95)%比(3.32±0.32)%,t=8.562,P<0.05]均显著提高(P<0.01),Caspase-3(4.93±0.50比1.32±0.13,t=6.325,P<0.05)、Caspase-9(4.46±0.45比1.45±0.14,t=6.105,P<0.05)活性显著提高(P<0.01),bax表达量上调(0.08±0.01比0.98±0.08,t=6.923,P<0.05)、bcl-2表达量下调(0.45±0.05比0.04±0.00,t=5.124,P<0.05),PI3K(0.16±0.01比2.23±0.24,t=4.236,P<0.05)、p-Akt(0.35±0.03比0.98±0.10,t=6.359,P<0.05)表达量下调;与Aβ25-35组比较,槲皮苷低、中、高剂量组细胞活力显著上升(0.432±0.043比0.362±0.036,t=4.259,P<0.05;0.464±0.046比0.362±0.036,t=5.856,P<0.05;0.523±0.052比0.362±0.036,t=6.258,P<0.05),细胞早期凋亡率[(12.19±1.22)%比(17.89±1.81)%,t=4.355,P<0.05;(6.69±0.68)%比(17.89±1.81)%,t=4.698,P<0.05;(3.31±0.37)%比(17.89±1.81)%,t=5.632,P<0.05]及晚期凋亡率[(12.46±1.25)%比(19.56±1.95)%,t=5.321,P<0.05;(8.37±0.84)%比(19.56±1.95)%,t=5.967,P<0.05;(4.28±0.43)%比(19.56±1.95)%,t=6.597,P<0.05]均显著降低,Caspase-3(4.06±0.41比4.93±0.50,t=5.326,P<0.05;3.08±0.32比4.93±0.50,t=6.257,P<0.05;2.08±0.21比4.93±0.50,t=6.984,P<0.05)、Caspase-9活性显著降低(3.52±0.35比4.46±0.45,t=4.214,P<0.05;2.78±0.27比4.46±0.45,t=5.987,P<0.05;1.85±0.18比4.46±0.45,t=6.865,P<0.05,bcl-2表达量上调(0.18±0.02比0.08±0.01,t=4.21Objective To explore effect of Quercitrin on PC12 cell injury indued byβ-amyloid protein25-35(Aβ25-35).Methods PC12 cells were randomly divided into normal control group,Aβ25-35 group,Quercitrin low,medium and high concentration group.Cell viability was measured by methyl thiazol tetrazolium(MTT)assay.Cell apoptosis was measured by flow cytometry.The activity of Caspase-3 and Caspase-9 was detected by colorimetry.The expression of B-cell lymphoma/leukemia-2(bcl-2),bcl-2-related X protein(bax),the activation of phosphatidylinositol-3-hydroxylkinase/protein kinase B(PI3K/Akt)signal pathway was detected by Western blotting.Results Compared with normal group,cell viability was reduced(0.362±0.036 vs.0.453±0.045,t=4.230,P<0.05),cell early apoptotic rate[(17.89±1.81)%vs.(2.31±0.23)%,t=8.623,P<0.05]and late apoptotic rate[(19.56±1.95)%vs.(3.32±0.32)%,t=8.562,P<0.05],Caspase-3(4.93±0.50 vs.1.32±0.13,t=6.325,P<0.05)and Caspase-9(4.46±0.45 vs.1.45±0.14,t=6.105,P<0.05)activity was increased(P<0.01),the expression of bax(0.08±0.01 vs.0.98±0.08,t=6.923,P<0.05)was up-regulated(P<0.01),the expression of bcl-2(0.45±0.05 vs.0.04±0.00,t=5.124,P<0.05;0.16±0.01 vs.2.23±0.24,t=4.236,P<0.05),p-Akt(0.35±0.03 vs.0.98±0.10,t=6.359,P<0.05)was down-regulated in Aβ25-35 group(P<0.01).Compared with Aβ25-35 group,cell viability was increased(0.432±0.043 vs.0.362±0.036,t=4.259,P<0.05;0.464±0.046 vs.0.362±0.036,t=5.856,P<0.05;0.523±0.052 vs.0.362±0.036,t=6.258,P<0.05),cell early apoptotic rate[(12.19±1.22)%vs.(17.89±1.81)%,t=4.355,P<0.05;(6.69±0.68)%vs.(17.89±1.81)%,t=4.698,P<0.05;(3.31±0.37)%vs.(17.89±1.81)%,t=5.632,P<0.05]and late apoptotic rate[(12.46±1.25)%vs.(19.56±1.95)%,t=5.321,P<0.05;(8.37±0.84)%vs.(19.56±1.95)%,t=5.967,P<0.05;(4.28±0.43)%vs.(19.56±1.95)%,t=6.597,P<0.05],Caspase-3(4.06±0.41 vs.4.93±0.50,t=5.326,P<0.05;3.08±0.32 vs.4.93±0.50,t=6.257,P<0.05;2.08±0.21 vs.4.93±0.50,t=6.984,P<0.05)and Caspase-9(3.52±0.35 vs.4.46±0.45,t=4.214,P<0.05;2.78±0.27 vs.4.46±0.45,t=5.987,P<0.05;1.85�

关 键 词：槲皮苷 PC12细胞 凋亡 磷脂酰肌醇-3-羟激酶/蛋白激酶B信号通路 

分 类 号：R285.5[医药卫生—中药学]