Quantitative analysis of the 5-lipoxygenase mRNA in the rat peritoneal macrophages cultured in vitro by capillary electrophoresis
- VernacularTitle:毛细管电泳定量分析在大鼠腹腔巨噬细胞中5-脂氧合酶信使核糖核酸表达中的应用
- Author:
Jieping LI
;
Tong LIANG
;
Keyuan ZHOU
- Publication Type:Journal Article
- Keywords:
Arachidonate 5-Lipoxygenase;
Macrophage;
Electrophoresis,capillary
- From:
Chinese Journal of Laboratory Medicine
2001;0(01):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To quantify the mRNA of 5-lipoxygenase (5-LO) in the rat peritoneal macrophages cultured in vitro by capillary electrophoresis (CE). Methods The rat peritoneal macrophages were isolated and cultured for indicated time. The mRNA of 5-LO was detected by RT-PCR, and the products of RT-PCR were quantified by CE. Results The DNA fragments in the 100 bp DNA marker and the products of the RT-PCR were separated successfully by CE with the sieving buffer containing 1.8% hydroxypropylmethylcellulose (HPMC). It proved that there were no significant changes on the expressions of 5-LO mRNA when the cells were cultured for 72 h quantified by CE. The mRNA of 5-LO significantly decreased by almost 80% by CE with the cells cultured for 120 h in vitro.Conclusions The products of RT-PCR could be separated and quantified by CE directly.The 5-LO mRNA could express normally in the rat peritoneal macrophages for 72 h in vitro.
