摘要
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,20mmol/L浓度时达最大值(P<0.05);20mmol/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。
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.
出处
《中华临床医师杂志（电子版）》
CAS
2009年第7期50-53,共4页
Chinese Journal of Clinicians(Electronic Edition)
