• Archives of Plastic Surgery
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• v.32 no.6
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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

• The Journal of Korean Medicine
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• v.20 no.1 s.37
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• pp.161-171
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• 1999

This study was performed to investigate the effect of complex formula(CKRG) consisting of Panax ginseng Radix rubra Koreana. Ganoderma, Cinnamomi Cortex, Glycyrrhizae Radix and Laminariae Thallus on brain ischemia and injury such as KCN-induced brain injury, forced brain ischemia, pulmonary thrombosis. The results were summarized as follows: 1. CKRG extracts showed a decrease of the duration of KCN-induced coma and showcd an increase in life expectancy. 2. CKRG extracts showed a decrease of neurologic grade in hind limb but did not affect neurologic grades in fore limb. Also. CKRG extracts showed a significant decrease of brain ischemic area and edema in MCA occlusion, 3. CKRG extracts showed a protective effect on pulmonary thrombosis induced by collagen and epinephrine. These data suggested that CKRG extracts could be applied to the protection of brain ischemia and injury.

• The Korean Journal of Physiology and Pharmacology
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• v.4 no.6
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• pp.525-530
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• 2000

During reperfusion of skeletal muscle after ischemia, lipid mediators, mainly eicosanoids, are released and may have a role in the pathogenesis of reperfusion injury. To validate the role of eicosanoids in the ischemia-reperfusion induced functional deficits in skeletal muscle, we compared muscle edema and the changes of eicosanoid concentration in the rat hind limb after ischemia-reperfusion injury by application of tourniquet. After 4 hours of ischemia, reperfusion was established for 4 hours by releasing tourniquet. To assess tissue damage, edema, and wet/dry weight ratios were determined and the eicosanoid concnentrations were measured by the HPLC. The muscle edema and the release of cyclooxygenase metabolites were not induced by the ischemia itself rather they were significantly increased by reperfusion. Indomethacin treatment ameliorated limb edema and decreased the release of $6-keto-PGF_{1{\alpha}},$ thromboxane $B_2,$ and $PGE_2$ inducedby reperfusion. But the inhibitory effect of indomethacin on edema (35%) was relatively low than the inhibitory effect on release of cyclooxygenase metabolites (up to 69%) by reperfusion. These results support the view that cyclooxygenase products may play a significant role in the formation of muscle injury by ischemia-reperfusion and suggest that nonsteroidal antiinflammatory agents might be partially beneficial to the management of acute limb ischemia-reperfusion injury.

• Archives of Plastic Surgery
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• v.33 no.1
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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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• v.44 no.5
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• pp.384-389
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• 2017

Background The aim of this study was to investigate the effects of remote ischemic conditioning on ischemia-reperfusion injury in rat muscle flaps histopathologically and biochemically. Methods Thirty albino rats were divided into 5 groups. No procedure was performed in the rats in group 1, and only blood samples were taken. A gracilis muscle flap was elevated in all the other groups. Microclamps were applied to the vascular pedicle for 4 hours in order to achieve tissue ischemia. In group 2, no additional procedure was performed. In groups 3, 4, and 5, the right hind limb was used and 3 cycles of ischemia-reperfusion for 5 minutes each (total, 30 minutes) was applied with a latex tourniquet (remote ischemic conditioning). In group 3, this procedure was performed before flap elevation (remote ischemic preconditoning). In group 4, the procedure was performed 4 hours after flap ischemia (remote ischemic postconditioning). In group 5, the procedure was performed after the flap was elevated, during the muscle flap ischemia episode (remote ischemic perconditioning). Results The histopathological damage score in all remote conditioning ischemia groups was lower than in the ischemic-reperfusion group. The lowest histopathological damage score was observed in group 5 (remote ischemic perconditioning). Conclusions The nitric oxide levels were higher in the blood samples obtained from the remote ischemic perconditioning group. This study showed the effectiveness of remote ischemic conditioning procedures and compared their usefulness for preventing ischemiareperfusion injury in muscle flaps.

• Journal of Haehwa Medicine
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• v.8 no.1
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• pp.425-449
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• 1999

For the evaluation of the effect on SWS, experiments were made on hyperlipidemia induced by hypercholesterol diet, inhibitory reaction to human platelet aggregation, Pulmonary thrombosis induced by collagen and epinephrine, global cerebral ischemia induced by KCN, brain ischemia induced by MCA occlusion, cytotoxicity of PC12 cells induced by amyloid ${\beta}$ protein(25-35), and NO production in RAW cells stimulated by lipopolysaccharide. The results were obtained as follows : 1. In the experiment on hyperlipidemia, the level of serum total cholesterol, phospholipid, and LDL-cholesterol were significantly decreased while the level of triglyceride, VLDL-cholesterol, and HDL-cholesterol had no significant change. 2. In the experiment on inhibitory reaction to platelet aggregation, SWS inhibited platelet aggregation induced by ADP(36.05%), by collagen(20.4%), and by thrombin(0.6%). 3. In the experiment on pulmonary thrombosis induced by collagen and epinephrine, the protective effect was found(37%). 4. In the experiment on global cerebral ischemia, coma duration induced by KCN changed insignificantly. 5. In the experiment on MCA occlusion, the change of neurologic grades on hind limb was significant only after the operation. Besides brain ischemic area and edema ratio were significantly decreased. 6. In the experiment on cytotoxicity of PC 12 cells induced by amyloid ${\beta}$ protein, the significant protective effect was found as concentration increases. 7. In the experiment on NO production in RAW cells stimulated by lipopolysaccharide, NO was significantly decreased. According to the results, it is expected that SWS might be effective on hyperlipidemia and brain damage.

• Journal of Haehwa Medicine
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• v.8 no.1
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• pp.379-401
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• 1999

The effect of KamiTongJonHaaATang extracts on hypercholesterolemia, platelet aggregation, pulm onary thrombosis, KCN-induced coma, forcal brain ischemia, cytotoxicity of PC12 cells induced by amyloid ${\beta}$ protein(25-35), and NO production in RAW cells stimulated lipopolysaccharide were investigated, respectively. The results were summarized as follows; 1. KTJHAT extracts showed a significant decrease of serum total cholesterol, triglyceride, phospholipid, LDL-cholesterol, and VLDL-cholesterol in hypercholesterolemia induced by 2% cholesterol diet in NZW rabbit. 2. KTJHAT extracts induced a significant inhibition of human platelet aggregation induced by thrombin and ADP but did not affect human platelet aggregation induced by collagen. 3. KTJHAT extracts showed a protective effect on pulmonary thrombosis induced by collagen and epinephrine. 4. KTJHAT extracts prolonged the duration of KCN-induced coma. 5. KTJHAT extracts showed a significant decrease of brain ischemic area and edema in MCA occlusion. Also, KTJHAT extracts showed a decrease of neurologic grade in hind limb but did not affect neurologic grade in fore limb. 6. KTJHAT extracts showed a protective effect on cytotoxicity of PC 12 cells induced by amyloid ${\beta}$ protein(25-35) in a dose dependent manner. 7. KTJHAT extracts showed a significant decrease of NO production in RAW cells induced by lipopolysaccharide. These results suggested that KTJHAT extracts might be usefully applied for prevention and treatement of thrombosis and brain damage.

• The Journal of Korean Medicine
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• v.25 no.3
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• pp.32-44
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• 2004

Objectives : This study was undertaken to prove the effect of GSHT on the glutamate receptor, free radical and brain damage in rats subjected to brain ischemia Methods : Levels of cultured cortical neuron death caused by toxic chemicals were measured by LDH release assay. Neuroprotective effects of GSHT on brain tissues were examined in vivo by ischemic model of middle cerebral artery (MCA) occlusion. Results : GSHT showed significant inhibitory effect on LDH release induced by NMDA-kinate-Fe/sup 2+/. GSHT remarkably decreased coma duration time in a nonfatal dose of KCN and showed higher survival rate in a fatal dose. GSHT remarkably decreased ischemic area and edema induced by the MCA blood flow block. GSHT showed high improvement of forelimb and hind limb test after MCA occlusion in neurological examination. GSHT showed no significant change after MCA occlusion in pathological observation of the normal group. Conclusions : These results indicate that GSHT can be used to treat the brain damage caused by brain ischemia. Further study will be needed about the functional mechanism, etc.

• The Korean Journal of Physiology and Pharmacology
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• v.25 no.3
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• pp.239-249
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• 2021

The present study explored the therapeutic potential of hydrogen sulfide (H2S) in restoring aging-induced loss of cardioprotective effect of remote ischemic preconditioning (RIPC) along with the involvement of signaling pathways. The left hind limb was subjected to four short cycles of ischemia and reperfusion (IR) in young and aged male rats to induce RIPC. The hearts were subjected to IR injury on the Langendorff apparatus after 24 h of RIPC. The measurement of lactate dehydrogenase, creatine kinase and cardiac troponin served to assess the myocardial injury. The levels of H2S, cystathionine β-synthase (CBS), cystathionine γ-lyase (CSE), nuclear factor erythroid 2-related factor 2 (Nrf2), and hypoxia-inducible factor (HIF-1α) were also measured. There was a decrease in cardioprotection in RIPC-subjected old rats in comparison to young rats along with a reduction in the myocardial levels of H2S, CBS, CSE, HIF-1α, and nuclear: cytoplasmic Nrf2 ratio. Supplementation with sodium hydrogen sulfide (NaHS, an H2S donor) and l-cysteine (H2S precursor) restored the cardioprotective actions of RIPC in old hearts. It increased the levels of H2S, HIF-1α, and Nrf2 ratio without affecting CBS and CSE. YC-1 (HIF-1α antagonist) abolished the effects of NaHS and l-cysteine in RIPC-subjected old rats by decreasing the Nrf2 ratio and HIF-1α levels, without altering H2S. The late phase of cardioprotection of RIPC involves an increase in the activity of H2S biosynthetic enzymes, which increases the levels of H2S to upregulate HIF-1α and Nrf2. H2S has the potential to restore aging-induced loss of cardioprotective effects of RIPC by upregulating HIF-1α/Nrf2 signaling.

• Microbiology and Biotechnology Letters
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• v.39 no.4
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• pp.364-373
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• 2011

Human embryonic stem cells have been highlighted as a valuable cellular source in the regenerative medicine field, due to their pluripotency. However, there is the challenge of the establishment of specific functional cell type forms of undifferentiated human embryonic stem cells (hESC). To establish and purify functional cell types from hESCs, we differentiated undifferentiated hESCs into vascular lineage cells and sorted the specific cell population from the whole cell population, depending on their cell volume, and compared them with the non-sorted cell population. We observed that about 10% of the PECAM positive population existed in the VEGF induced differentiating human embryoid body (hEB), and differentiated hEBs were made into single cells for cell transplantation. After making single cells, we performed cell sorting using a fluorescence-activated cell sorter (FACs), according to their cell volume on the basis of FSC region gating, and compared their therapeutic capacity with the non-sorted cell population through cell transplantation into hindlimb ischemic disease model mice. 4 Weeks after cell transplantation, the recovery rate of blood perfusion reached 54% and 17% in the FSC regions of sorted cells- and non-sorted cells, respectively. This result suggests that derivation of a functional cell population from hESCs can be performed through cell sorting on the basis of cell volume after preliminary differentiation induction. This approach may then greatly contribute to overcoming the limitations of marker sorting.