摘要
目的探讨瑞芬太尼对缺氧复氧的人肝细胞损伤的作用及其可能的作用机制。方法将培养的人肝细胞分为5组,分别为正常对照(C)组、单纯缺氧复氧损伤(AR)组、瑞芬太尼(R)组、白屈菜红碱(CH)组和混合(R+CH)组,n=6。C组正常培养,其余各组缺氧培养15h,复氧5h,然后收集细胞,进行:①肝细胞线粒体的丙二醛(MDA)含量测定;②用Annexin V/PI染色,于流式细胞仪下计数凋亡细胞的百分数;③提取RNA,以PKC mRNA为目的基因,进行RT-PCR,然后电泳,对条带的吸光值进行比较。结果AR组的MDA含量明显高于C组,R组和CH组的MDA含量介于AR与C组之间(P<0.05)。AR组的肝细胞凋亡率明显高于C组,R组和CH组的肝细胞凋亡率介于AR与C组之间(P<0.05)。AR组的PKC mRNA转录量明显高于C组,R组和CH组的PKC mRNA转录量介于AR与C组之间(P<0.05),R+CH组的PKC mRNA转录量低于C组(P<0.05)。结论瑞芬太尼对缺氧复氧的肝细胞具有保护作用,且可能与其抑制PKC mRNA的转录有关。
Objective To assess the protective effect of remifentanil on cultured human hepatocytes against anoxia-reoxygenation injury. Methods Cultured hepatocytes were divided into 5 groups： group C receiving normoxia as control; groups AR, R, CH, R ＋ CH receiving 15-hour xypoxia followed by 5-hour reoxygenation （group R receiving 5 ng/ml remifentanil, group CH 10 μmol/L chelerythrine, group R ＋ CH 5 ng/ml remifentanil and 10 μmol/L chelerythrine before reoxygenation）. The content of MDA in the hepatocyte mitochondria were measured. The rate of apoptotic cells was measured by flow cytometry. The expression of protein kinase C mRNA was measured by RT-PCR. Results Anoxia-reoxygenation caused dramatic increase in the content of MDA, the rate of apoptotic cells and the expression of protein kinase C mRNA. The three indexes mentioned above of groups R and CH were between that of groups C and AR （P 〈 0. 05 ）. The expression of protein kinase C mRNA in group R ＋ CH was less than that in group C （P 〈 0.05 ）. Conclusion Remifentanil can protect cultured hepatocytes against anoxia-reoxygenation injury, and its mechanism may be associated with protein kinase C mRNA.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2007年第20期1976-1978,共3页
Journal of Third Military Medical University
关键词
瑞芬太尼
肝细胞
缺氧
复氧
蛋白激酶C
remifetanil
hepatocytes
anoxia
reoxygenation
protein kinase C
