• 대한약리학회지
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• 제27권2호
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• pp.109-118
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• 1991

허혈심근의 재관류시 arachidonic acid가 반응성 산소대사물의 발생원으로서 심근 손상에 미치는 영향을 검토하였다. Langendorff 관류장치를 이용하여 흰쥐 적출심장을 0.5ml/min의 저용량으로 관류 (45분)한 후 정상관류 (7 ml/min)로 복귀 시키므로써 실험적인 허혈-재관류 심장을 만들었다. 재관류시 Na arachidonate $(10^{-7}{\sim}10^{-2}{\mu}g/ml)$를 투여한 후 superoxide anion 생성을 관찰하고, 심근 손상의 지표로 lactic dehydrogenase(LDH)유리를 측정 하였으며 이들에 대한 각종 arachidonic acid 대사 억제 약물의 영향을 비교 검토하였다. Superoxide anion 생성은 SOD-억제성 ferricytochrorme C 환원 반응을 이용하였다. 연구성적은 다음과 같다. 1) 저용량 관류후 재관류시 ferricytochrorme C환원은 superoxide dismutase (SOD, 300 U/ml) 및 indomethacin (60 nmole/ml), ibuprofen $(30\;{\mu}g/ml)$에 의하여 억제되었다. 2) Na arachidonate는 용량의존적으로 ferricytochrorme C 환원을 증가 시켰으며 반응성 산소대사물 제거효소인 superoxide dismutase (SOD, 300 U/ml)에 의하여 현저히 억제되었다. 3) Na arachidonate $(10^{-3}\;{\mu}g/ml)$에 의한 superoxide anion 생성은 cyclooxygenase 억제약물인 indomethacin (60 nmol/ml), lipooxygenase 억제약물인 nordihydroguaiaretic acid$(NDGA,\;0.1\;{\mu}mole/ml)$, arachidonic acid의 substrate inhibitor인 eicosatetraynoic acid $(ETYA,\;1\;{\mu}g/ml)$에 의하여 현저히 억제되었다. 4) Na arachidonate는 LDH 유리를 증가시켰으며 SOD에 의하여 유의하게 억제 되었다. 5) Na archidonate에 의한 LDH 유리증가는 indomethacin, NDGA, ETYA에 의하여 유의하게 억제 되었다. 이상의 결과로 흰쥐의 허혈-재관류심근에서 arachidonic acid는 그 대사 과정에서 반응성 산소대사물을 발생하고 이는 심근세포손상에 부분적으로 기여할 수 있을 것으로 여겨졌다.

• Journal of Chest Surgery
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• 제29권2호
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• pp.199-207
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• 1996

Adenosine을 함유한 심정지액의 심근 보호 효과심장수술에 있어서 허혈성 심정지는 거의 필연적인 과정이며 이로 인한 재관류후의 심근 손상은 심장 수술의 성패를 결정하는 가장 중요한 요소중의 하나이다. Adenosine은 강력한 혈관 확장제이며 칼슘통로에 길항하고 칼릅통로를 활성화 시컴으로 빠른 심정지를 유도하여 허혈성 심정지후 회복을 향상시킨다. Adenosine이 칼륨보다 빠른 심정지를 유도하고 허혈성 심정지후 혈역학적 회복을 향상시키는데 효과적이라는 가설을 검정하기위해 횐쥐의 심장을 Langendorff 장치에 연결하고 Krebs-Henseleit 완충액으로 관류 시킨후 심근 마비액을 이용하여 60분간 심정지를 시켰으며 심근 마비액은 구성에 따라 3군으로 분류하였다. A군(n=10), 칼륨을 함유하지 않은 St. Thomas 심정지액에 adenosine 10mmole/L 을 첨가. B군(n: 10), 칼륨을 함유한다. Thomas 심정지액에 adenosine 400 mol/L 을 첨가. C군(n=10), 기존의 St. Thomas 심정지액 Adenosine 을 함유한군(A군과 B군)은 대조군(C군)에 비해 빠른 심정지를 유발하였다(p<0.01). 관상동맥 관류량은 대조군과 비교하여 A군에서는 재관류 20분과 30분에 B군에서는 재관류 20분에 증가가 있었다(p<0.01). 수축기 동맥압은 A군과 B군에서 재관류 10분에 향상이 있었다(p<0.01). dpfdt는 A군에서 재관류 10분에 증가가 있었다(p<0.05). A군과 B군은 대동맥 관류량, 심박출량, 심박수에서도 C군에 비해 좋은 회복율을 보였으나 통계학적 유의성은 없었다. CPK 치는 A군에서 낮게 측정 되었다(p<0.01). 이상의 결과로 보아adenosine을 함유한 심정지액이 기존의 칼륨 심정지액에 비하여 빠른 심정지를 유도하였고 심장의 회복에 더욱 유리한 결과를 보여 주었다.

• Journal of Chest Surgery
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• 제31권9호
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• pp.837-844
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• 1998

배경: 아데노신은 심근의 허혈상태에서 심근세포로부터 분비되어 부정맥과 심근허혈 및 수술후 재관류손상을 억제시키는 호르몬으로 알려져 있다. 아데노신의 심근보호 효과에 대한 연구는 주로 심정지액 속에 포함시킨 아데노신의 효과에 대하여 이루어 지고 있으나, 심정지액 속에 포함된 아데노신의 적정용량에 대해서는 보고가 다양하다. 저자들은 자체제작한 심폐체외순환 모델을 이용하여 단일용량의 아데노신(0.75 mg/Kg/min)이 우수한 심근보호효과를 나타낸 결과를 보고한 바 있으나 적절한 용량이었는지에 대한 확신은 없다. 따라서 본 연구의 목적은 심정지액 속에 포함된 아데노신의 적정용량을 알아보는데 있다. 대상 및 방법: 연구방법은 쥐를 이용하여 심정지시 심정지액(St. Thomas 심정지액)에 첨가한 아데노신의 용량에 따라 1군(0.5 mg/Kg/min), 2군(0.75 mg/Kg/min) 및 3군(1 mg/Kg/min) 으로 나누어 각각 10마리씩 실험하여 비교하였다. 마취 후 적출된 쥐심장의 대동맥과 좌심방에 도관을 삽관한 후에 심폐체외순환 모델에 연결하여 비작업성 순환과 작업성 순환을 시행하면서 혈역학적 수치를 측정하여 이를 대조값으로 이용하였다. 심정지액을 주입하여 심정지를 유도한 후에 90분간 허혈상태로 유지한 다음 비작업성 순환을 시행 후 작업성 순환으로 바꿔 10분, 30분 및 60분에 혈역학적 수치(심박동수, 수축기 대동맥압, 1분간 대동맥 박출량 및 관동맥관류량)를 측정하고, 생화학적 검사(CPK, Lactic Acid) 및 심장의 수분함유량도 측정하였다. 측정된 수치는 심정지 전 측정한 대조값에 대한 백분율로 환산하여 비교하였다. 결과: 실험 결과 심정지 전에 측정한 대조값 사이에는 세군 사이의 통계적인 유의성이 없었다. 심정지액의 주입 후 3군에서 가장 빨리 심정지가 일어났으며(p<0.05), 재관류 후 심박동이 돌아온 시간은 1군과 2군이 3군에 비하여 통계학적으로 유의하게 심박동이 빨리 돌아 왔다 (p<0.05). 그러나 1군과 2군 사이에는 유의성이 없었다. 심장의 재관류 후 측정한 심박동수의 회복률에서 10분에 측정한 값은 세군 사이에 유의성이 없었으나, 30분과 60분 에 측정한 값은 1군이 2군과 3군에 비하여 유의하게 높았으며(p<0.05), 2군도 3군에 비하여 유의하게 높았다(p<0.05). 수축기 대동맥압의 회복률, 1분 동안의 대동맥 박출량 및 심박출량(1분 동안의 대동맥 박출량과 관동맥관류량을 합산한 값)은 10분, 30분 및 60분에서 모두 2군이 1군과 3군에 비하여, 1군은 3군에 비하여 유의하게 높았다(p<0.05). 관동맥관류량의 회복률은 10분과 30분에 측정한 값은 2군이 1군과 3군에 비하여, 1군은 3군에 비하여 유의하게 높았으며(p<0.05), 60분에 측정한 값은 1군과 2군이 3군에 비하여 유의하게 높았다(p<0.05). 심근의 수분함유량과 관상동맥 관류량의 생화학적 검사결과 CPK와 Lactic Acid는 세군 사이에 유의성이 없었다. 결론 : 이상의 결과로 아데노신을 심정지용액에 첨가시 비교적 적정 용량은 0.75 mg/Kg/min 을 투여하는 것이 적절할 것으로 생각된다.

• The Korean Journal of Physiology and Pharmacology
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• 제20권2호
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• pp.185-192
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• 2016

Ampicillin, a ${\beta}$-lactam antibiotic, dose-dependently protects neurons against ischemic brain injury. The present study was performed to investigate the neuroprotective mechanism of ampicillin in a mouse model of transient global forebrain ischemia. Male C57BL/6 mice were anesthetized with halothane and subjected to bilateral common carotid artery occlusion for 40 min. Before transient forebrain ischemia, ampicillin (200 mg/kg, intraperitoneally [i.p.]) or penicillin G (6,000 U/kg or 20,000 U/kg, i.p.) was administered daily for 5 days. The pretreatment with ampicillin but not with penicillin G significantly attenuated neuronal damage in the hippocampal CA1 subfield. Mechanistically, the increased activity of matrix metalloproteinases (MMPs) following forebrain ischemia was also attenuated by ampicillin treatment. In addition, the ampicillin treatment reversed increased immunoreactivities to glial fibrillary acidic protein and isolectin B4, markers of astrocytes and microglia, respectively. Furthermore, the ampicillin treatment significantly increased the level of glutamate transporter-1, and dihydrokainic acid (DHK, 10 mg/kg, i.p.), an inhibitor of glutamate transporter-1 (GLT-1), reversed the neuroprotective effect of ampicillin. Taken together, these data indicate that ampicillin provides neuroprotection against ischemia-reperfusion brain injury, possibly through inducing the GLT-1 protein and inhibiting the activity of MMP in the mouse hippocampus.

• Journal of Korean Neurosurgical Society
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• 제40권2호
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• pp.103-109
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• 2006

Objective : Activated endothelial cells mediate the cascade of reactions in response to hypoxia for adaptation to the stress. It has been suggested that hypoxia, by itself, without reperfusion, can activate the endothelial cells and initiate complex responses. In this study, we investigated whether hypoxia-induced endothelial products alter the endothelial permeability and have a direct cytotoxic effect on nerve cells. Methods : Hypoxic condition of primary human umbilical vein endothelial cells[HUVEC] was induced by $CoCl_2$ treatment in culture medium. Cell growth was evaluated by 3,4,5-dimethyl thiazole-3,5-diphenyl tetrazolium bromide [MTT] assay Hypoxia-induced products [$IL-1{\beta},\;TGF-{\beta}1,\;IFN-{\gamma},\;TNF-{\alpha}$, IL-10, IL-6, IL-8, MCP-l and VEGF] were assessed by enzyme-linked immunosorbent assay. Endothelial permeability was evaluated by Western blotting. Results : Prolonged hypoxia caused endothelial cells to secrete IL -6, IL -8, MCP-1 and VEGF. However, the levels of IL -1, IL -10, $TNF-{\alpha},\;TGF-{\beta},\;IFN-{\gamma}$ and nitric oxide remained unchanged over 48 h hypoxia. Hypoxic exposure to endothelial cells induced the time-dependent down regulation of the expression of cadherin and catenin protein. The conditioned medium taken from hypoxic HUVECs had the cytotoxic effect selectively on neuroblastoma cells, but not on astroglioma cells. Conclusion : These results suggest the possibility that endothelial cell derived cytokines or other secreted products with the increased endothelial permeability might directly contribute to nerve cell injury followed by hypoxia.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 2004년도 Annual Meeting of KSAP : New Drug Development from Natural Products
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• pp.3-10
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• 2004

Oxidative stress is a constant threat to all living organisms and an immense repertoire of cellular defense systems is being employed by most pro- and eukaryotic systems to eliminate or to attenuate oxidative stress. Ischemia and reperfusion is characterized by both a significant oxidative stress and characteristic changes in the antioxidant defense. Heme oxigenase-l (HO-l) is up-regulated by various stimuli including oxidative stress so that it is thought to participate in general cellular defense mechanisms against ischemic injury in mammalian cells. Higenamine, an active ingredient of Aconite tuber, has been shown to have antioxidant activity along with inhibitory action of inducible nitric oxide synthase (iNOS) expression in various cells. In the present study, we investigated whether higenamine and related analogs protect cells from oxidative cellular injuries by modulating antioxidant enzymes, such as HO-l, MnSOD etc. R-form of YS-51 was the most potent inducer of HO-l in bovine endothelial cells, which inhibited apoptotic cell death by H$_2$O$_2$. HO-1 induction by YS 51 was mediated by PI3 kinase activation in which PKA- as well as PKG pathway is considered as important regulators. YS-51 also induced Mn-SOD mRNA expression by activating c-jun N-terminal kinase in endothelial cells and Hela cells. In ROS 17/2.1 cells, higenamine and enetiomers of related compounds inhibited iNOS expression by cytokine mixtures. Taken together, higenamine and related compounds can be developed as possible protective agents from oxidative cell injury or death.

• Biomolecules & Therapeutics
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• 제8권2호
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• pp.160-166
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• 2000

The present study examined influence of various ischemic duration on extent of focal ischemic brain injury induced by middle cerebral artery occlusion (MCAO) in rats. The MCAO was produced by insertion of a 17 mm silicone-coated 4-0 nylon surgical thread to the origin of MCA through the internal carotid artery for 30, 60, 90, 120 min (transient) or 24 hr (permanent) in male Sprague-Dawley rats under isoflurane anesthesia. Reperfusion in transient MCAO models was achieved by pulling the thread out of the internal carotid artery. Only rats showing neurological deficits characterized by left hemiparesis and/or circling to the left, were included in cerebral ischemic groups. The rats were sacrificed 24 hr after MCAO and seven serial coronal slices of the brain were stained with 2,3,5-triphenyltetrazolium chloride. Infarct size was measured using a computerized image analyzer. Ischemic damage was common in the frontoparietal cortex (somatosensory area) and the lateral segment of the striatum while damage to the medial segment of the striatum depended on the duration of the occlusion. In the 30-min MCAO grouts, however, infarcted region was primarily confined to the striatum and it was difficult to clearly delineate the region since there was mixed population of live and dead cells in the nucleus. Infarct volume was generally increased depending on the duration of MCAO, showing the most severe damage in the permanent MCAO group. However, there was no significant difference in infarct size between the 90-min and 120-min MCAO groups. % Edema also tended to increase depending on the duration of MCAO. The results suggest that the various focal ischemic rat models established in the present study can be used to evaluate in vivo neuroprotective activities of candidate compounds or to elucidate pathophysiological mechanisms of ischemic neuronal cell death.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 2003년도 Annual Meeting of KSAP : International Symposium on Pharmaceutical and Biomedical Sciences on Obesity
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• pp.62-62
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• 2003

It is now convincing that free radical generation is involved in the pathophy siological mechanisms of ischemic stroke, particularly in ischemia-reperfusion injury. The present study, therefore, examined neuroprotective effect of aloesin isolated from Aloe vera, which was known to have antioxidative activity, in a rat model of transient focal cerebral ischemia. Transient focal cerebral ischemia was induced by occlusion of middle cerebral artery for 2 hr with a silicone-coated 4-0 nylon monofilament in male Sprague-Dawley rats under isoflurane anesthesia Aloesin (1, 3, 10, 30 and 50 mg/kg/injection) was administered intravenously 3 times at 0.5, 2 and 4 hr after onset of ischemia. Neurological score was measured 24 hr after onset of ischemia immediately before sacrifice. Seven serial coronal slices of the brain were stained with 2,3,5-triphenyltetrazolium chloride and infarct size was measured using a computerized image analyzer. Treatment with the close of 1 or 50 mg/kg did not significantly reduce infarct volume compared with the saline vehicle-treated control group. However, treatments with the closes of 3 and 10 mg/kg significantly reduced both infarct volume and edema by approximately 47% compared with the control group, producing remarkable behavioral recovery effect. Treatment with the close of 30 mg/kg also significantly reduced infarct volume to a lesser extent by approximately 33% compared with the control group, but produced similar degree of behavioral recovery effect. In addition, general pharmacological studies showed that aloesin was a quite safe compound. The results suggest that aloesin can serve as a lead chemical for the development of neuroprotective agents by providing neuroprotection against focal ischemic neuronal injury.

• Journal of Chest Surgery
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• 제20권2호
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• pp.230-240
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• 1987

Cardioplegia and myocardial protection were performed under cardiopulmonary bypass during open-heart surgery with the use of cold St. Thomas Hospital cardioplegic solution [4=C] for the coronary artery perfusion and normal saline solution [4- C] for the topical cardiac cooling. To maintain the state of myocardial protection, coronary artery reperfusion was carried out using St. Thomas Hospital cardioplegic solution at the interval of 30 minutes. A total number of patients studied were 57 cases, including 37 cases of correction for congenital cardiac anomalies and 20 cases for acquired heart valvular diseases. Cardiopulmonary bypass time during the surgery was observed to be average of 87.89*47.55 hours, aortic cross-clamping time to be average of 76.68~44.27 hours raging from 30 to 191 minutes. In order to evaluate the effects of myocardial protection in the surgery, serum enzyme levels were determined. To observe the relationship between aortic cross-clamping time and myocardial protection effects, patients studied were divided into the following 3 groups. I group: aortic cross-clamping time, 60 minutes, II group: aortic cross-clamping time, 90 minutes, III group: aortic cross-clamping time, over 91 minutes. 1. Changes in serum enzyme levels in postoperative period. [1] SCOT; The postoperative value [increased over 200 units] for ischemic myocardial injury during operation was observed in 11 cases [19.3% of the total] of the total patients studied, of which 4 cases [13.3%] in I group, 1 case [10.0%] in II group, and 6 cases [35.3%] in III group. [2] LDH; The positive value [increased over 900 units] for ischemic myocardial injury during operation was observed in 9 cases [15.7% of the total] of the total patients studied, of which 2 cases [6.6%] in I group, 1 case [10.0%] in II group, and 6 cases [35.3%] in III group. [3] CPK; The positive value [increased over 800 units] for ischemic myocardial injury during operation was observed in 10 cases [17. 5% of the total] of the total patients studied, including 4 cases [13. 3%] in I group, 1 case [10.0%] in II group, and 5 cases [29.4%] in III group. 2. The myocardial protection method used in the present study was demonstrated to be effective for the myocardial protection in the surgery with aortic cross-clamping time of up to 90 minutes. A few ischemic myocardial injury were observed in the surgery with aortic cross-clamping time over 91 minutes, but no significant cardiac dysfunction was noted. The surgery with aortic cross-clamping time of up to 191 minutes did not appear to give rise any significant interference with postoperative recovery.

• Journal of Chest Surgery
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• 제22권2호
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• pp.225-233
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• 1989

Cardioplegia and myocardial protection were performed under cardiopulmonary bypass during open heart surgery with the use of St. Thomas Hospital cardioplegic solution [4 [C] for the coronary artery perfusion and normal saline solution [4[ c] for the topical cardiac cooling. To maintain the state of myocardial protection, coronary artery reperfusion was carried out using St. Thomas Hospital cardioplegic solution at the interval of 30 minutes. A total number of patients studied were 57 cases, including 37 cases of correction for congenital anomalies and 20 cases for acquired heart diseases. Cardiopulmonary bypass time during the surgery was observed to be average of 87.89*47.55 hours, aortic cross-clamping time [ACCT] to be average of 76.68*44.27 hours raging from 30 to 191 minutes. In order to evaluate the effects of myocardial protection in the surgery, serum enzyme levels were determined. To observe the relationship between ACCT and myocardial protection effects, patients studied were divided into the following 3 groups. I group: ACCT 60 minutes, II group: ACCT 90 minutes, III group: ACCT over 91 minutes [1] SGOT; The positive value [increased over 200 units] for ischemic myocardial injury during operation was observed in 11 cases [19.3% of the total] of the total patients studied, of which 4 cases [13.3%] in I group, 1 case [10.0%] in II group, and 6 cases [35.3%] in III group. [2] LDH; The positive value [increased over 900 units] for ischemic myocardial injury during operation was observed in 9 cases [15.7% of the total] of the total patients studied, of which 2 cases [6.6%] in I group, 1 case [10.0%] in II group and 6 cases [35.3%] in III group. [3] CPK; The positive value [increased over 800 units] for ischemic myocardial injury during operation was observed in 10 cases [17.5% of the total] of the total patients studied, including 4 cases [13.3%] in I group, 1 case [10.0%] in II group, and 5 cases [29.4%] in III group [4] The myocardial protection method used in the present study was demonstrated to be effective for the myocardial protection in the surgery with ACCT of up to 90 minutes. A few ischemic myocardial injury were observed in the surgery with ACCT over 91 minutes, but no significant cardiac dysfunction was noted. The surgery with ACCT of up to 191 minutes did not appear to give rise any significant interference with postoperative recovery.