中华临床医师杂志（电子版）
中華臨床醫師雜誌（電子版）
중화림상의사잡지（전자판）
CHINESE JOURNAL OF CLINICIANS(ELECTRONIC VERSION)
2009年
7期
1135-1141
,共7页
董晓丽%刘京升%金戈%明海霞%李海龙
董曉麗%劉京升%金戈%明海霞%李海龍
동효려%류경승%금과%명해하%리해룡
阿尔茨海默病%肌肽%过氧化氢%疾病模型,动物%半胱氨酸天冬氨酸蛋白酶3%细胞凋亡
阿爾茨海默病%肌肽%過氧化氫%疾病模型,動物%半胱氨痠天鼕氨痠蛋白酶3%細胞凋亡
아이자해묵병%기태%과양화경%질병모형,동물%반광안산천동안산단백매3%세포조망
Alzheimer disease%Carnosine%Hydrogen peroxide%Disease models,animal%Caspase 3%Apoptosis
目的 探讨L-肌肽对H2O2诱导大鼠肾上腺嗜铬细胞瘤细胞(PC12细胞)凋亡保护的作用机制.方法 在H2O2诱导PC12细胞凋亡模型的基础上加入肌肽,采用MTT比色法检测肌肽对H2O2抑制PC12细胞增殖的影响;RT-PCR法检测凋亡相关基因Caspase-3 mRNA的表达变化;免疫组织化学SABC法检测Caspase-3蛋白的表达,流式细胞仪检测细胞凋亡.结果 不同浓度的肌肽对H2O2损伤的PC12细胞的存活率有显著的提高作用,20 mmol/L浓度时达最大值(P<0.05);20 mmol/L的肌肽作用于PC12细胞可降低其Caspase-3 mRNA的表达及Caspase-3蛋白的表达,流式细胞仪检测显示,肌肽可抑制细胞的早期和晚期凋亡.结论 肌肽对H2O2损伤的PC12细胞有保护作用,机制可能是通过抑制Caspase-3的表达来抑制PC12细胞的凋亡而实现的.
目的 探討L-肌肽對H2O2誘導大鼠腎上腺嗜鉻細胞瘤細胞(PC12細胞)凋亡保護的作用機製.方法 在H2O2誘導PC12細胞凋亡模型的基礎上加入肌肽,採用MTT比色法檢測肌肽對H2O2抑製PC12細胞增殖的影響;RT-PCR法檢測凋亡相關基因Caspase-3 mRNA的錶達變化;免疫組織化學SABC法檢測Caspase-3蛋白的錶達,流式細胞儀檢測細胞凋亡.結果 不同濃度的肌肽對H2O2損傷的PC12細胞的存活率有顯著的提高作用,20 mmol/L濃度時達最大值(P<0.05);20 mmol/L的肌肽作用于PC12細胞可降低其Caspase-3 mRNA的錶達及Caspase-3蛋白的錶達,流式細胞儀檢測顯示,肌肽可抑製細胞的早期和晚期凋亡.結論 肌肽對H2O2損傷的PC12細胞有保護作用,機製可能是通過抑製Caspase-3的錶達來抑製PC12細胞的凋亡而實現的.
목적 탐토L-기태대H2O2유도대서신상선기락세포류세포(PC12세포)조망보호적작용궤제.방법 재H2O2유도PC12세포조망모형적기출상가입기태,채용MTT비색법검측기태대H2O2억제PC12세포증식적영향;RT-PCR법검측조망상관기인Caspase-3 mRNA적표체변화;면역조직화학SABC법검측Caspase-3단백적표체,류식세포의검측세포조망.결과 불동농도적기태대H2O2손상적PC12세포적존활솔유현저적제고작용,20 mmol/L농도시체최대치(P<0.05);20 mmol/L적기태작용우PC12세포가강저기Caspase-3 mRNA적표체급Caspase-3단백적표체,류식세포의검측현시,기태가억제세포적조기화만기조망.결론 기태대H2O2손상적PC12세포유보호작용,궤제가능시통과억제Caspase-3적표체래억제PC12세포적조망이실현적.
Objective To study protective effect of L-carnosine on apoptosis of PC12 cells induced by hydrogen peroxide(H2O2).Methods Based on the model of apoptosis of PC12 cells induced by H2O2 with 200 μmol/L,and then the models were treated with L-carnosine at different concentrations.Cell viability was tested by MTT assay.The mRNA and protein expressions of Caspase-3 were tested by means of reverse transcription-polymerase chain reaction(RT-PCR)and immunocytochemical technique.The apoptosis was detected by flow cytometry(FCM).Results Pretreatment with different concentrations of L-carnosine(10,20,30 mmol/L)for 30 min increased the survival rate of PC12 cells,the peak was 20 mmol/L(P<0.05),pretreatment with 20 mmol/L of L-carnosine decreased the expressions of Caspase-3 mRNA and protein,decreased apoptosis.Conclusions L-carnosine can prevent PC12 cell from H2O2 injury.Neurotoxicity mechanism of protection is likely related to decreasing Caspase-3,and prevent cell from apoptosis.
