• 한국임상수의학회지
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• 제27권5호
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• pp.533-539
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• 2010

최근 줄기세포를 이용한 재생의학분야의 세포치료 연구가 활발히 진행되고 있으나 상대적으로 복잡한 구조를 갖는 신장에서는 난치성 질환인 급성신부전에 대한 줄기세포 치료의 효과는 아직도 논란의 여지가 많다. 이는 연구자마다 다양한 동물모델을 사용하는 것이 한 원인으로 지적되고 있다. 본 연구에서는 줄기세포의 치료효과 분석을 위한 최적의 동물모델 조건을 확립하고자 한다. 쥐를 사용하여 세 종류의 급성신장손상모델을 다음과 같이 다양한 농도와 조건에서 분석하였다: glycerol (대조군, 8 ml/kg, 10 ml/kg), cisplatin (대조군, 5 mg/kg, 10 mg/kg)과 허혈/재관류 (대조군, 35 분 허혈, 45 분 허혈). 줄기세포의 재생능력 평가를 위한 모델의 적합성 정도를 신장손상과 회복의 형태적, 기능적 패턴과 생존률 분석을 통하여 실시하였다. 독성모델인 glycerol과 cisplatin은 각각 10 ml/kg과 7.5 mg/kg 농도에서, 그리고 허혈모델은 45 분 모델에서 대조군 및 다른 비교모델에 비해 장기간의 관찰 기간 동안 신기능의 유의적 차이를 보였다. 또한 45 분 허혈 모델에서 50%의 사망률을 보인 것을 제외하고는 모든 군에서 관찰기간 동안 100% 생존율을 보였다. 조직소견에서 허혈과 glycerol 모델에서는 2일째, cisplatin 모델에서는 5일째 각각 유의적인 조직변화가 관찰되었다. 따라서, 관찰기간 동안 급격한 생존율 감소 없이 안전하고, 조직학적 평가와 신기능에서 대조군과 유의적인 차이를 보이는 cisplatin 7.5 mg/kg, glycerol 10 ml/kg, 35 분 허혈 모델이 치료효과를 평가하기에 적합한 모델로 사료되며, 줄기세포의 세포치료효과 분석에 적합한 급성신장손상 모델로 추천된다.

• Biomolecules & Therapeutics
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• 제7권4호
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• pp.354-361
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• 1999

In this study, the effects of tauroursodeoxycholic acid (TUDCA) on ischemia/ reperfusion injury were investigated on isolated heart perfusion models. Hezrts were perfused with oxygenated Krebs-henseleit solution (pH 7.4, $37^{\cire}C$) on a Langendorff apparatus. After equilibration, isolated hearts were treated with TUDCA 100 and 200 $\mu\textrm{M}$ or vehicle (0.02% DMSO) for 10 min before the onset of ischemia in single treatment group. In 7 day pretreatment group. TUDCA 50, 100 and 200 mg/kg body weight were given orally for 7 days before operation. After global ischemia (30 min), ischemic hearts were reperfused for 30 min. The physiological (i.e. heart rate, left ventricdular developed pressure, coronary flow, double product, time to contracture formation) and biochemical (lactate dehydrogenase; LDH) parameters were evaluated. In vehicle-treated group, time to contracture formation was 810 sec during ischemia, LVDP was 34.0 mmHg at the endpoint of reperfusion and LDH activity in total reperfusion effluent was 34.3 U/L. Single treatment with TUDCA did not change the postischemic recovery of cardiac function, LDH and time to contractur compared with ischemic control group. TUDCA pretreatment showed the tendency to decrease LDH release and to increase time to contracture and coronary flow. Our findings suggest that TUDCA does not ameliorate ischemia/reperfusion-reduced myocardial damage.

• Maxillofacial Plastic and Reconstructive Surgery
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• 제28권4호
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• pp.287-294
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• 2006

Vascular thrombosis and ischemic necrosis still remain the most significant threats to the survival of free flaps. To date, neutrophils have been implicated in the pathogenesis of postischemic injury. Several studies have demonstrated that modulating the neutrophil response to ischemia-reperfusion injury can decrease the extent of the injury. In addition, some authors noticed that mast cell counts were also increased in flaps exposed to state of ischemia/reperfusion. So, we designed to evaluate the role of mast cells in ischemia/reperfusion by blocking histamine and to compare the effect of L-arginine, a nitric oxide precursor which is known to prevent neutrophil-mediated tissue injury. Epigastric island skin flaps were elevated in 30 rats and rendered ischemic. Thirty minutes prior to reperfusion, the rats were treated with intraperitoneal saline, diphenhydramine, cimetidine, and L-arginine. The necrosis rate of flap at 7 days, the number of neutrophils and mast cells at 20 hours were evaluated. In conclusion, histamine receptor blockers as well as L-arginine significantly decreased flap necrosis in a rat skin island ischemia-reperfusion flap model, but the protective effect was not significantly different in both agent groups.

• Archives of Plastic Surgery
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• 제33권1호
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• pp.31-38
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• 2006

The effect of melatonin on morphological changes after ischemia-reperfusion injury was investigated in rat skeletal muscle. Dimethyl-sulfoxide(DMSO) was also tested for comparison. Muscle injury was evaluated in 4 groups as a single laparotomy group(control), ischemia-reperfusion group, DMSO group, melatonin group. Left hind limb ischemia was induced for 4 hours by vascular clamping of the common femoral artery and followed by 24 hours of reperfusion. The midportion of gastrocnemius muscle was taken for histological evaluation. In light microscopic study, ischemia-reperfusion group showed severe neutrophil infiltration, interstitial edema, and partial loss or degeneration of muscle fibers. The muscle tissue of melatonin group showed relatively normal architecture with mild inflammatory cell infiltration. In electron microscopic study, dilated cisternae of sarcoplasmic reticulum, dilated mitochondria with electron loose matrix and dilated cristae, disordered or loss of myofilament, indistinct A-band and I-band, intracytoplasmic vacuoles, and markedly decreased glycogen granules were observed in ischemia-reperfusion group. But relatively well maintained A-band, I-band, Z-line, M-line, and mildly dilated mitochondria with well preserved cristae were observed in melatonin group. The DMSO group showed intermediately attenuated ultrastructural changes. The results show that melatonin improves morphologically ischemia-reperfusion injury more effectively than DMSO. In conclusion, melatonin seems to be a promising agent that can salvage the skeletal muscle from severe ischemia-reperfusion injury.

• Archives of Plastic Surgery
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• 제32권6호
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• pp.683-688
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• 2005

The ischemia-reperfusion injury of the skeletal muscles is caused by generation of reactive oxygen during ischemia and reperfusion. Melatonin or N-Acetyl-5-methoxy- tryptamine is suggested to have antioxidant effects in several tissues. In present study, we examined the protective effect of melatonin in a rat hind limb ischemia-reperfusion injury. Dimethyl-sulfoxide(DMSO) was also tested for comparison. Ischemia was induced for 4 hours by vascular clamping and followed by 1 hour or 24 hours of reperfusion. Muscle injury was evaluated in 4 groups such as single laparotomy group(control), ischemia-reperfusion group, DMSO group, melatonin group. Eedema ratio and malondialdehyde(MDA) of muscle tissue and serum level of creatine kinase(CK), were measeured at the end of reperfusion. DMSO and melatonin group showed significant amelioration of edema and serum CK compared with ischemia-reperfusion group. The decreasing effect was more prominent in melatonin group. The muscle tissue MDA concentration is significantly lower in melatonin group than in ischemia-reperfusion group. The results show that melatonin prevents and improves ischemia-reperfusion injury more effectively in a rat hind limb than DMSO dose. Thus, clinically the melatonin may be used for a beneficial treatment of such injuries

• 약학회지
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• 제56권4호
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• pp.260-267
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• 2012

This study examined the effects of baicalin, a bioactive flavonoid isolated from Scutellaria baicalensis, on hepatic injury caused by ischemia/reperfusion (I/R) in alcoholic fatty liver. Rats were fed an ethanol liquid diet or a control isocaloric diet for 5 weeks, and then subjected to 60 min of hepatic ischemia and 5 h of reperfusion. Baicalin (200 mg/kg) was administered intraperitoneally 24 and 1 h before ischemia. After reperfusion, baicalin attenuated the increase in serum alanine aminotransferase activity. The levels of cytosolic cytochrome c protein expression, caspase-3 activity, the number of apoptotic cells increased after reperfusion, which were higher in ethanol-fed animals, were attenuated by baicalin. Following I/R, the hepatic lipid peroxidation was elevated, whereas hepatic glutathione content was decreased. These changes attenuated by baicalin. In ethanol-fed animals, baicalin augmented the increases in heme oxygenase-1 protein and mRNA expressions, and nuclear Nrf2 expression. In conclusion, our findings suggest that baicalin ameliorates I/R-induced hepatocellular damage by suppressing apoptosis and oxidative stress in alcoholic fatty liver.

• Archives of Reconstructive Microsurgery
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• 제21권1호
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• pp.61-67
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• 2012

Purpose: The purpose of this study was to investigate the role of mast cells and their product, histamine and leukotriene in ischemia-reperfusion injury. Methods: Forty Sprague-Dawley rats were divided into four groups. (Group I: Control group without ischemia, Group II: Normal saline with ischemia, Group III: Cimetidine with ischemia, Group IV: Zafirlukast with ischemia) Skin flap was elevated and ischemic insult was given by clamping the artery for 12 hours. Before reperfusion, the rats were treated with saline, cimetidine and zafirlukast. Flap survival was evaluated at 7 days. Neutrophil counts, mast cell counts were evaluated 24 hours after reperfusion. Results: Flap survival rate in the control group was 92.33%, whereas normal saline group had 37.34% survivals. Cimetidine and zafirlukast treated group showed significantly higher survival rates than normal saline group. The neutrophil and mast cell counts in cimetidine and zafirlukast treated group were significantly decreased than normal saline group. Cimetidine treated group showed higher survival rate and lower cell counts than zafirlukast treated group. Conclusion: The administration of cimetidine and zafirlukast can decrease neutrophils and mast cells caused by ischemia-reperfusion and increase flap survivals. It is suggests that antihistamine and leukotriene receptor antagonist have protective effect against ischemia-reperfusion injury to skin flap in rat.

• Journal of Chest Surgery
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• 제28권11호
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• pp.963-966
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• 1995

Ischemia reperfusion injury occurs in various diseases. The role of oxygen free radicals in IR injury of the lung has been spotlighted and many studies have been performed. In this study, we tried to prepare a stable rat lung model for IR injury, focusing on surrounding conditions as hilar stripped left lung, clamped left pulmonary artery and bronchus,and declamped after determined period was passed, and right main pulmonary aretery was clamped. Arterial blood gas analyes were performed at 1, 10, 20, 30, minutes after reperfusion. Before clamping, PaO2 was 95 to 120 mmHg in all animals. There were six groups; Group I : temperature 15o C, and 120 minutes clamping, Group II: 20 oC, and 120 minutes clamping, Group III : 25 oC, and 120 minutes clamping, Group IV : 15oC, 90 minutes clamping, Group V : 20 oC, 90 minutes clamping,Group VI: 20 oC, 75 minutes clamping. Each groups contained 10 Sprague Dayley rats. The humidity was maintained 100 % as circulation imerged isotonic Hartmann`s solution of the pleural cavity. In group IV, V, and VI, PaO2 decreased significantly in all animals immediately after reperfusion, but 43 % survived till 10 minutes after reperfusion, it was 74.0$\pm$5.7, 73.3$\pm$10.8,and 88.2$\pm$17.7 mmHg. Pulmonary edema was observed histologically in 2/10 animals in group IV, 6/10 in group V , 3/10 in group VI, 9/10 in group I, and the other lungs showed all edema. We established a stable model by setting ischemic time,and temperature, between 75 to 90 minutes,15 to 20o C, and isotemperature Hartmann`s solution immersion of the pleural cavity.

• Journal of Chest Surgery
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• 제30권9호
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• pp.847-853
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• 1997

Although the effects of adenosine on the heart, including the clinical suppression of cardiac arrhythmias, have been recognized for more than half a century, it is only in the last decade that the therapeutic potential of adenosine has been recognized. The objective of this study was to determine if augmentation of myocardial adenosine levels during global ischemia improves functional recovery after reperfusion. We used to modified Langendonf system to evaluate myocardial protective effect. Isolated rat hearts were subjected to 90 minutes of deep hypothermic arrest(15$^{\circ}C$) with modified St. Thomas'Hospital cardioplegic solution used to provide myocardial protection. Myocardial adenosine levels were augmented during ischemia by providing exogenous adenosine in the cardioplegic solution. Two groups of hearts w re studied: (1) control group(n=10) cardioplegia alone; (2) adenosine group(n=10) adenosine(0.75mg/kg/min) added to the cardioplegic solution. Significantly better percent recovery(p<0.01) in hemodynamics(except heart rate) at 60 minutes after reperfusion was evident compared to baseline values in the adenosine group. (systolic no란ic pressure : 78.5$\pm$3.6% vs 66.6$\pm$5.9%, airtic overflow volume : 61.7$\pm$ 11.6% vs 37.2$\pm$ 15.4%, coronary flow volume 77.1$\pm$7.5% vs 57.2$\pm$ 11.1%, and cardiac output : 65.6$\pm$ 11.5% vs 44.2$\pm$ 12.4%). Heart rate was similar in two groups(94.4$\pm$4.8% vs 95.3 $\pm$ 6.8%). Adenosine groups resulted in significantly rapid recovery time of heart beat after reperEusion(p<0.01) (24.5$\pm$7.6 sec. vs 179.0$\pm$ 131.1sec.). In biochemical study, CPK levels(0.1 $\pm$0.3U/L vs 1.4$\pm$0.8U/L) and lactic acid levels(0.08$\pm$0.Immol/L vs 0.34$\pm$0.2 mmol/L) were significantly low in adenosine groups(p<0.01). We concluded that adenosine included cardioplegia have better recovery effects after r perfusion in myocardial ischemia compared to adenosine free cardioplegia.

• Journal of Korean Neurosurgical Society
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• 제30권1호
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• pp.12-19
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• 2001

Objective : Albumin is a very useful drug for the improving of cerebral blood volume and the oncotic effect in cerebral ischemia or cerebral vasospasm. The purpose of this study was to examine the morphological and neurological effect of albumin therapy on reperfusion injury following transient focal cerebral ischemia. Materials and Methods : 18 Male Sprague-Dawley rats weighing 270-320g were used. The ischemia model was produced by 2-hour period of transient middle cerebral artery occlusion with a poly-L-lysin coated intraluminal suture. The agent(20% human serum albumin[HSA]) or control solution(NaCl 0.9%) was administered intravenously at a dosage of 1% of body weight immediate after reperfusion following a 2-hour period occlusion. Neurological function was evaluated by the postural reflex and the forlimb placing test during occlusion(at 60 min) and daily for 3 days thereafter. The brain was perfusion-fixed, and infarct volumes and brain edema were measured. Results : The HSA significantly improved the neurological score in treated group. The rats of albumin treatment group showed significantly reduced total infarct volume(by 34%) and brain edema(by 81%) compared with salinetreated rats. Conclusion : HSA showed a substantial effect on the transient focal cerebral ischemia and reperfusion injury model. These results may indicate its usefulness in treating reperfusion injury patients after thrombolysis treatment for the thrombo-embolic major cerebral artery occlusions.