반복적인 허혈양상화가 흰쥐의 앞정강근과 가자미근에서 산화질소합성효소발현에 미치는 영향
Effects of Repetitive Ischemic Preconditioning on the Expression of Nitric Oxide Synthasein Tibialis Anterior and Soleus Muscles of the Rat
2
조회
0
다운로드
2
인용
0
참고문헌
키워드
초록
산화질소합성효소(nitiric oxide synthase, NOS)는 산화질소 생성에 필요한 효소로, 허혈손상에 영향을 준 다고 알려져 있다. 이에 본 연구자들은 흰쥐의 백색근육과 적색근육섬유에서 허혈양상화가 산화질소합성효소의 발현에 미치는 영향을 관찰하고자 하였다. 9주령의 SD계 수컷 흰쥐를 대조군과 허혈양상화 3회, 6회, 10회군으로 나누었고, 재관류 0시간, 3시간, 6시간, 24 시간, 72시간군으로 세분하였다. 허혈양상화는 흰쥐의 온엉덩동맥을 5분 허혈과 5분 재관류를 반복 실시하였고, 재 관류 시간에 따라 실험동물을 희생시켜 앞정강근과 가자미근을 적출하였다. nNOS와 iNOS, eNOS를 관찰하기 위 해 Western blot analysis와 면역조직화학염색을 시행하여 다음과 같은 결과를 얻었다. 허혈양상화 후 nNOS는 대조군보다 증가되었고, 앞정강근에서는 3회군과 6회군이 10회군보다 높게 관찰되었다. 허혈양상화 후 iNOS의 증가는 가자미근보다 앞정강근에서 더 높게 관찰되었고, 10회군에서 가장 높게 관찰되었 다. 허혈양상화 후 eNOS는 대조군보다 증가되었고, 앞정강근보다 가자미근에서 더 높게 관찰되었다. 재관류 0시간 과 3시간에 eNOS는 허혈양상화 6회군과 10회군에서 높게 관찰되었다. 이상의 실험결과로 허혈양상화는 nNOS와 iNOS, eNOS를 증가시키고, 10회의 허혈양상화는 iNOS를 증가시켜 허혈손상을 일으킬 것으로 생각되며, 앞정강근이 가자미근보다 허혈자극에 민감하게 반응한다고 판단되었다.
Abstract
Nitric oxide synthases (NOSs) that catalyzed the conversion of L-arginine to nitric oxide and L-citrulline play a role in ischemic-reperfusion injury. The purpose of this study was to observe the expression patterns of nNOS, iNOS and eNOS in the rat tibialis anterior and soleus muscles after multiple cyclic episodes of ischemic preconditioning (IP). Nine weeks old male SD rats were divided into control and IP groups. The IP group was further divided into 3 groups based on cycle of IP. For IP, left commom iliac artery was occluded 3, 6 and 10 times for 5 minutes ischemia followed by 5 minutes reperfusion using rodent vascular clamps. The animals were sacrificed at 0, 3, 6, 24 and 72 hours of reperfusion and the left tibialis anterior and soleus muscles were removed. The expression of nNOS, iNOS and eNOS were examined with immunohistochemical methods and Western blot analysis. IP increased the expression of nNOS, compared with the control. In the tibialis anterior muscle, the levels of nNOS in the 3IP and 6IP were higher than that in 10IP. IP increased the expression of iNOS, compared with the control, and the levels of iNOS in tibialis anterior muscle were higher than that in soleus muscle. The level of iNOS in the 10IP was higher than those in the 3IP and 6IP. IP increased the expression of eNOS, compared with the control, and the level of eNOS in soleus muscle were higher than that in tibialis anterior muscle. At 0 and 3 hours after reperfusion, the level of eNOS in 6IP and 10IP were higer than that in 3IP. In summary, these results suggest that the ischemic preconditioning increases the expression of nNOS, iNOS and eNOS, and 10 times of ischemic preconditioning may induce ischemic injury through upregulation of iNOS. And tibialis anterior muscle is more susceptabile to ischemic injury than soleus muscle.
참고문헌 (30)
- Amstrong RB, "Muscle fiber type composition of the rat hindlimb. Am J Anat 171", pp.259~272 (1984)
- Balon TW, "Evidence that nitric oxide increases glucose transport in skeletal muscle", 82, pp.359~363 (1997)
- Blaisdell FW, "The pathophysiology of skeletal muscle ischemia and the reperfusion syndrome: a review", 10, pp.620~630 (2002)
- Geller DA, "Molecular biology of nitric oxide synthase", 17, pp.7~23 (1998)
- Granger DL, "requirement for the microbiostaticactivity of murine macrophages. J Clin Invest 81", specificami(acid), pp.1129~1136 (1988)
- Gurke L, "Function of fast- and slow-twitch rat skeletal muscle following ischemia and reperfusion at different intramuscular temperatures", 32, pp.135~141 (2000)
- Hussain SN, "Expression of nitric oxide synthases and GTP cyclohydrolase I in the ventilatory and limb muscles during endotoxemia", 17, pp.173~180 (1997)
- Elander A, "Purinemetabolism after in vivo ischemia and reperfusion inrat skeletal muscle. Am J Physiol 258", pp.~1678 (1990)
- Jefayri MK, "Attenuation of reperfusioninjury by renal ischaemic preconditioningAttenuation of reperfusion injury by renal ischaemic preconditioning the role of nitric oxide", 85, pp.1007~1013 (2000)
- Jugdutt BI, "Nitric oxide and cardioprotection during ischemia-reperfusion", 7, pp.391~405 (2002)
- Kane AJ, "Inducible nitric oxide synthase (iNOS) activity promotes ischaemic skin flap survival", 132, pp.1631~1638 (2001)
- Lochner A, "Nitric oxide triggers classic ischemic preconditoning", 962, pp.402~414 (2002)
- Mattei A, "Preconditioning with short cycles improves ischemic tolerance in rat fast- and slow-twitch skeletal muscle", 32, pp.297~304 (2000)
- Mungrue IN, "Cardiomyocyte overexpression of iNOS in mice results in peroxynitrite generation, heart block, and sudden death", 109, pp.735~743 (2002)
- Murry CE, "a delay of lethal cell injury in ischemic myocardium.Circulation 74", pp.1124~1136 (1986)
- Muscari C, "Early preconditioning prevents the loss of endothelial nitric oxide synthase and enhances its activity in the ischemic", reperfused rat heart, 2004, pp.74, 1127~1137
- Pang CY, "Pharmacologic interventionin schemia -induced reperfusion injury in the skeletalmuscle. Microsurgery 14", pp.176~182 (1993)
- Punkt K, "Nitric oxide synthase isoforms I, III and protein kinase-Ctheta in skeletal muscle fibres of normal and streptozotocin-induced diabetic rats with and without Ginkgo biloba extract treatment", 33, pp.213~219 (2001)
- Punkt K, "Myopathy-dependent changes in activity of ATPase, SDH and GPDH and NOS expression in the different fibre types of hamster muscles", 104, pp.15~22 (2002)
- Reid BM, "Role of nitric oxide in skeletal muscle: synthesis, distribution and functional importance", 162, pp.401~409 (1998)
- Rubinstein I, "Involvement of nitric oxide system in experimental muscle crush injury", 101, pp.1325~1333 (1998)
- Rudnick J, "Differential expression of nitric oxide synthases (NOS 1-3) in human skeletal muscle following exercise countermeasure during 12 weeks of bed rest", 18, pp.1228~1230 (2004)
- Sakamoto K, "Inducible nitric oxide synthase inhibitors abolished histological protection by late ischemic preconditioning in rat retina", 82, pp.512~518 (2006)
- Seo YK, "Age-related alterations of apoptosis in ischemic-reperfused rat tibialis anterior and soleus muscles", 16, pp.291~299 (2003)
- Adelman B, "Skeletal muscle fiber typedoes not predict sensitivity to postischemic damage. J SurgRes 53", pp.535~541 (1992)
- Stewart AG, "Physiological andpathophysiological roles of nitric oxide. Microsurgery 15", pp.693~702 (1994)
- Tidball JG, "Mechanical loading regulates NOS expression and activity in developing and adult skeletal muscle", 275, pp.C260~266 (1998)
- Welter R, "The effects of nitric oxide on electron transport complexes", 331, pp.914~ (1996)
- Woitaske MD, "Effects of fiber type on ischemia-reperfusion injury in mouse skeletal muscle", 102, pp.2052~2063 (1998)
- Zhang L, "Reperfusion injury is reduced in skeletal muscle by inhibition of inducible nitric oxide synthase", 94, pp.1473~1478 (2003)