Effects of age and ischemia on cardiolipin measured by silicic acid high-performance liquid chromatography in isolated rat hearts
- VernacularTitle:高效液相色谱法研究年龄和缺血对大鼠心肌线粒体心磷脂含量的影响
- Author:
Haijun SHAO
;
Jingyuan LI
;
Zhumin SHAO
;
Jianwei FAN
;
Yinming ZENG
- Publication Type:Journal Article
- Keywords:
cardiolipin;
HPLC;
lipid
- From:
Chinese Pharmacological Bulletin
1987;0(03):-
- CountryChina
- Language:Chinese
-
Abstract:
Aim To establish a method to quantitatively determine cardiolipin content and investigate effects of age and ischemia on cardiolipin in isolated rat hearts.Methods Male SD rats, 4 months old,12 months old and 24 months old were used throughout the experiment. Each had 24 ones,which were randomly distributed to control,ischemia 30 min and ischemia 40 min groups(n=8). Control group hearts were perfused for a total of 70 min and ischemia groups hearts were perfused for a 20 min equilibration period,and then underwent 30 or 40 min of no-flow ischemia followed by reperfusion for a 20 min period, respectively.Cardiolipin was quantitatively determined by high-performance liquid chromatography following extraction of lipids from subsarcolemmal mitochondria(SSM)and interfibrillar mitochondria(IFM).Results Under the condition of HPLC,the retention time of cardiolipin was 13.092 min. The standard curve was Y^ (?V?s) =20 877 455X(mmol?L-1)-352 028(r=0.9993) with a validated quantitation range of 0.2～3.2 mmol?L-1,and the lowest concentration of detection was 0.005 mmol?L-1 (S/N=3). At the concentration of 0.2,0.8,3.2 mmol?L-1,absolute recovery rates ranged from 0.8505 to 0.9519 and relative recovery rates ranged from 0.9898 to 1.0429. The RSD of the precision within-day and between-day was less than 0.11. Cardiolipin content in SSM of nonischemic group of 24 months old rats was lower than that of both 4 months old and 12 months old, but no differences among them were observed in IFM.In all age rats, compared with control group,cardiolipin contents of SSM from ischemia 30 min and ischemia 40 min groups were lower, and effects of ischemia 40 min are not pronounced; there were no differences in CL contents of IFM among three groups.Conclusions The assay method was shown to be sensitive, suitable and accurate for determination of cardiolipin located in mitochondria from perfused hearts of rats. Myocardial ischemia and older age decreased cardiolipin content in SSM, and had no remarkable effects on cardiolipin in IFM.
