Analysis of Nucleotides and Their Derivatives in Renal Tissue of Rat during Ischemia by HPLC

흰쥐의 신허혈에서 HPLC를 이용한 핵산대사산물의 분석

  • Kim, Seong-Yong (Department of Biochemistry College of Medicine, Yeungnam University) ;
  • Kim, Jung-Hye (Department of Biochemistry College of Medicine, Yeungnam University)
  • 김성용 (영남대학교 의과대학 생화학교실) ;
  • 김정희 (영남대학교 의과대학 생화학교실)
  • Published : 1992.06.30

Abstract

In rat kidney, the changes in concentrations of nucleotides and their derivatives during ischemia induced by renal artery ligation was measured quantitatively with high performance liquid chromatography(HPLC). After the ligation of renal artery for 60minutes, the concentrations of the nucleotides and derivatives were measured. In ischemic tissue, IDP was significantly decreased from $217.4{\pm}12.68{\mu}g$ in control to $80.7{\pm}18.39{\mu}g$ (p<0.01) ; ATP, $307.2{\pm}56.63{\mu}g$ to $47.6{\pm}5.95{\mu}g$ (p<0.01) ; ADP+AMP, $227.1{\pm}7.98{\mu}g$ to $61.4{\pm}3.92{\mu}g$(P<0.01); $NAD^+$, $217.9{\pm}4.49{\mu}g$ to $126.6{\pm}10.44{\mu}g$(P<0.01) ; GTP, $202.5{\pm}23.76{\mu}g$ to $117.7{\pm}14.24{\mu}g$ (P<0.05) ; GMP, $54.5{\pm}9.03{\mu}g$ to $23.7{\pm}0.46{\mu}g$(p<0.05), and inosine, $16.6{\pm}3.45{\mu}g$ to $7.8{\pm}0.87{\mu}g$ (P<0.05). But hypoxanthine and xanthine were significantly increased from $113.0{\pm}15.58{\mu}g$ to $159.7{\pm}12.07{\mu}g$ (P<0.05) and from $87.7{\pm}6.77{\mu}g$ to $173.1{\pm}12.52{\mu}g$ (P<0.01). In ischemic kidney, concentration of ATP was decreased to 39.9% of control at 10 minutes, 19.8% at 30 minutes, and 15.5% at 60 minutes, and ADP+AMP were decreased to 70.3% of control at 10 minutes, 67.3% at 30 minutes, and to 27.0% at 60 minutes, but hypoxanthine and xanthine were increased to 121.5% and 127.1% at 10 minutes, 126.0% and 174.4% at 30 minutes, and 141.4% and 197.3% at 60 minutes Total adenosine nucleotides were decreased to 20.3% of control during 60 minutes of ischemia, but hypoxanthine and xathine were increased to 157.5 % of control. These results suggest that the changes in the concentration of nucleotides and their metabolic derivatives are useful indices of the extents of tissue ischemia in rat kidney.

흰쥐 신동맥 결찰로 허혈을 유도하여 허혈상태의 핵산 및 핵산 대사산물의 변화가 정상조직과 어떠한 차이가 있는지를 알아 보기 위하여 HPLC(high performance liquid chromatography)를 이용하여 nucleotides를 분리하고 분석 및 정량한 결과이다. 60분간의 허혈후 핵산 및 핵산 대사산물의 변화를 보면 조직 1 gram당의 IDP농도는 대조군 $217.4{\pm}12.68{\mu}g$에서 $80.7{\pm}18.39{\mu}g$으로(P<0.01), ATP는 $307.2{\pm}56.63{\mu}g$에서 $47.6{\pm}5.95{\mu}g$으로(P<0.01), ADP+AMP는 $227.1{\pm}7.98{\mu}g$에서 $61.4{\pm}3.92{\mu}g$으로(P<0.01), $NAD^+$ $217.9{\pm}4.49{\mu}g$에서 $126.6{\pm}10.44{\mu}g$으로(P<0.01), GTP는 $202.5{\pm}23.76{\mu}g$에서 $117.7{\pm}14.24{\mu}g$으로(P<0.05), GMP는 $54.5{\pm}9.03{\mu}g$에서 $23.7{\pm}0.46{\mu}g$으로(P<0.05), 그리고 inosine은 $16.6{\pm}3.45{\mu}g$에서 $7.8{\pm}0.87{\mu}g$으로(P<0.05) 유의하게 감소한 반면, hypoxanthine은 $113.0{\pm}15.58{\mu}g$에서 $159.7{\pm}12.07{\mu}g$으로(P<0.05), xanthine은 $87.7{\pm}6.77{\mu}g$에서 $173.1{\pm}12.52{\mu}g$으로(P<0.01) 유의하게 증가하였다. 허혈의 지속시간에 따른 변화는 ATP가 10분에 대조군의 39.9%, 30분에 19.8% 그리고 60분에 15.5%로 감소하였으며, ADP+AMP는 10분에 70.3%, 30분에 67.3%, 60분에 27.0%로 감소하는 경향이었고, hypoxanthine과 xanthine은 각각 10분에 121.5%와 127.1%로, 30분에 126.0%와 174.4%로, 그리고 60분에 141.4%와 197.3%로 증가하는 경향이었다. 허혈 60분동안 고 에너지원인 adenosine nucleotide의 총량은 대조군의 20.3%로 감소하였고, purine nucleotide의 대사산물인 hypoxanthine과 xanthine의 총량은 157.5%로 증가하였다. 이상의 결과로 보아 쥐의 허혈 신조직에서의 핵산 및 그 대사산물의 농도는 허혈의 정도를 알 수 있는 유용한 지표라고 생각된다.

Keywords