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地塞米松和淀粉样β蛋白片段25-35对PC12细胞损伤和凋亡的影响
引用本文:李维祖,张文,李侠,尹艳艳,李卫平. 地塞米松和淀粉样β蛋白片段25-35对PC12细胞损伤和凋亡的影响[J]. 中国药理学与毒理学杂志, 2011, 25(5): 430-435. DOI: 10.3867/j.issn.1000-3002.2011.05.004
作者姓名:李维祖  张文  李侠  尹艳艳  李卫平
作者单位:1. 安徽医科大学药理学教研室,抗炎免疫药理学省部共建教育部重点实验室,国家中医药管理局中药药理三级实验室,安徽合肥230032
2. 皖南医学院弋矶山医院药剂科,安徽芜湖,230000
3. 安徽医科大学药理学教研室,抗炎免疫药理学省部共建教育部重点实验室,国家中医药管理局中药药理三级实验室,安徽合肥230032;安庆医药高等专科学校,安徽安庆246003
基金项目:安徽省高校省学术带头人科学研究资助,安徽省人才基金,安徽省教育厅自然科学项目,安徽省教育厅自然科学项目
摘    要:目的探讨地塞米松(DEX)和淀粉样β蛋白片段25-35(Aβ25-35)联合作用对PC12细胞损伤和凋亡的影响。方法采用单独或联合应用DEX 0.1~10μmol.L-1和Aβ25-35 1~5μmol.L-1作用PC12细胞24 h,MTT法测定细胞活力;膜联蛋白-Ⅴ,PI双染流式细胞仪检测细胞凋亡率,PI单染流式细胞仪检测细胞凋亡峰;逆转录-PCR检测胱天蛋白酶3 mRNA的表达水平。结果 MTT结果显示,与正常对照组相比,DEX 5和10μmol.L-1,Aβ25-35 1,5和10μmol.L-1,DEX 5+Aβ25-35 1,5和10μmol.L-1及DEX 10+Aβ25-35 5,10μmol.L-1组均可明显降低PC12细胞存活率,而DEX联合Aβ25-35能明显减少PC12细胞数(P<0.01)。流式细胞仪结果显示,DEX 5μmol.L-1和Aβ25-35 1μmol.L-1单独作用对PC12细胞凋亡率和亚二倍体凋亡峰有一定的增加作用(P<0.05),两者联合作用能明显增加PC12细胞早期和中晚期凋亡率,增加亚二倍体凋亡峰(P<0.01);RT-PCR结果显示,DEX 5μmol.L-1和Aβ25-35 1μmol.L-1单独作用对PC12细胞胱天蛋白酶3 mRNA的表达水平没有明显影响,它们联合作用能明显增加PC12细胞胱天蛋白酶3mRNA的表达水平(P<0.05)。结论 DEX和Aβ25-35联合作用能明显增加对PC12细胞的损伤,促进胱天蛋白酶3 mRNA表达,诱导PC12细胞凋亡。

关 键 词:地塞米松  淀粉样β蛋白  阿尔茨海默病  细胞凋亡
收稿时间:2010-12-06

Effect of dexamethasone and amyloid beta protein fragment 25-35 on PC12 cells injury and apoptosis
LI Wei-zu,ZHANG Wen,LI Xia,YIN Yan-yan,LI Wei-ping. Effect of dexamethasone and amyloid beta protein fragment 25-35 on PC12 cells injury and apoptosis[J]. Chinese Journal of Pharmacology and Toxicology, 2011, 25(5): 430-435. DOI: 10.3867/j.issn.1000-3002.2011.05.004
Authors:LI Wei-zu  ZHANG Wen  LI Xia  YIN Yan-yan  LI Wei-ping
Affiliation:LI Wei-zu1,ZHANG Wen2,LI Xia1,YIN Yan-yan1,LI Wei-ping1,3(1.Department of Pharmacology,Key Laboratory of Anti-inflammatory and Immunopharmacology,Ministry of Education,Key Laboratory of Chinese Medicine Research and Development,State Administration of Traditional Chinese Medicine,Anhui Medical University,Hefei 230032,China,2.Department of Pharmacy,Yijishan Hospital,Wannan Medical University,Wuhu 241001,3.Anqing Medical and Pharmaceutical College,Anqing 246003,China)
Abstract:OBJECTIVE To study the effect of dexamethasone (DEX) and amyloid beta protein fragment 25-35 (Aβ25-35) on PC12 cells injury and apoptosis. METHODS DEX 0.1-10 μmol·L-1 and Aβ25-35 1-5 μmol·L-1 were administered alone or in combination to PC12 cells. PC12 cells survival was detected by MTT assay. PC12 cells apoptosis was detected by flow cytometer with Annexin- Ⅴ/PI double stain and PI single stain. Caspase 3 mRNA level in PC12 cells was detected by RT-PCR. RESULTS MTT results showed that, DEX 5, 10 μmol·L-1, Aβ25-35 1, 5, 10 μmol·L-1, DEX 10+Aβ25-35 1, 5, 10 μmol·L-1 and DEX 5+Aβ25-35 5 and 10 μmol·L-1 could decrease PC12 cells survival after 24 h treatment. While combination of DEX with Aβ25-35 could significantly decrease PC12 cells number (P<0.01). DEX 5 μmol·L-1, Aβ25-351 μmol·L-1 could increase the apoptotic rate of PC12 cells. And DEX 5+Aβ25-35 1 μmol·L-1 could significantly increase the apoptotic rate of PC12 cells (P<0.01). Single DEX 5 or Aβ25-35 1 μmol·L-1 had no significant effect on caspase 3 mRNA levels, but DEX 5 μmol·L-1+Aβ25-35 1 μmol·L-1 could significantly increase caspase 3 mRNA levels. CONCLUSION Combination of DEX with Aβ25-35 can significantly increase the injury and apoptosis in PC12 cells.
Keywords:dexamethasone  amyloid beta protein  Alzheimer′s disease  apoptosis
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