• 대한수의학회지
• /
• 제57권1호
• /
• pp.1-7
• /
• 2017

Parkinson's disease (PD) is an irreversible neurological disorder with related locomotor dysfunction and is characterized by the selective loss of nigral neurons. PD can be experimentally induced by 6-hydroxydopamine (6-OHDA). It has been reported that reactive oxygen species, which deplete endogenous glutathione (GSH) levels, may play important roles in the dopaminergic cell death characteristic of PD. Fucoidan, a sulfated algal polysaccharide, exhibits anti-inflammatory and anti-oxidant actions. In this study, we investigated whether fucoidan can protect against 6-OHDA-mediated cytotoxicity in SH-SY5Y cells. Cytotoxicity was evaluated by using MTT and LDH assays. Fucoidan alleviated cell damage evoked by 6-OHDA dose-dependently. Fucoidan reduced the number of apoptotic nuclei and the extent of annexin-V-associated apoptosis, as revealed by DAPI staining and flow cytometry. Elevation of lipid peroxidation and caspase-3/7 activities induced by 6-OHDA was attenuated by fucoidan, which also protected against cytotoxicity evoked by buthionine-sulfoximine-mediated GSH depletion. Reduction in the glutathione/glutathione disulfide ratio induced by 6-OHDA was reversed by fucoidan, which also inhibited 6-OHDA-induced disruption of mitochondrial membrane potential. The results indicate that fucoidan may have protective action against 6-OHDA-mediated neurotoxicity by modulating oxidative injury and apoptosis through GSH depletion.

• The Korean Journal of Physiology and Pharmacology
• /
• 제15권2호
• /
• pp.83-88
• /
• 2011

A large body of evidence has indicated that induction of endogenous antioxidative proteins seems to be a reasonable strategy for delaying the progression of cell injury. In our previous study, cilostazol was found to increase the expression of the antioxidant enzyme heme oxygenase-1 (HO-1) in synovial cells. Thus, the present study was undertaken to examine whether cilostazol is able to counteract tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$)-induced cell death in endothelial cells via the induction of HO-1 expression. We exposed human umbilical vein endothelial cells (HUVECs) to TNF-${\alpha}$ (50 ng/ml), with or without cilostazol ($10{\mu}M$). Pretreatment with cilostazol markedly reduced TNF-${\alpha}$-induced viability loss in the HUVECs, which was reversed by zinc protoporphyrine IX (ZnPP), an inhibitor of HO-1. Moreover, cilostazol increased HO-1 protein and mRNA expression. Cilostazol-induced HO-1 induction was markedly attenuated not only by ZnPP but also by copper-protoporphyrin IX (CuPP). In an assay measuring peroxisome proliferator-activated receptor-${\gamma}$ (PPAR-${\gamma}$) transcription activity, cilostazol directly increased PPAR-${\gamma}$ transcriptional activity which was completely abolished by HO-1 inhibitor. Furthermore, increased PPAR-${\gamma}$ activity by cilostazol and rosiglitazone was completely abolished in cells transfected with HO-1 siRNA. Taken together, these results indicate that cilostazol up-regulates HO-1 and protects cells against TNF-${\alpha}$-induced endothelial cytotoxicity via a PPAR-${\gamma}$-dependent pathway.

• The Korean Journal of Physiology and Pharmacology
• /
• 제10권5호
• /
• pp.263-271
• /
• 2006

Since astrocytes were shown to play a central role in maintaining neuronal viability both under normal conditions and during stress such as ischemia, studies of the astrocytic response to stress are essential to understand many types of brain pathology. The micro array system permitted screening of large numbers of genes in biological or pathological processes. Therefore, the gene expression patterns in the in vitro model of astrocytes following exposure to oxygen-glucose deprivation (OGD) were evaluated by using the micro array analysis. Primary astrocytic cultures were prepared from postnatal Swiss Webster mice. The cells were exposed to OGD for 4 hrs at $37^{\circ}C$ prior to cell harvesting. From the cultured cells, we isolated mRNA, synthesized cDNA, converted to biotinylated cRNA and then reacted with GeneChips. The data were normalized and analyzed using dChip and GenMAPP tools. After 4 hrs exposure to OGD, 4 genes were increased more than 2 folds and 51 genes were decreased more than 2 folds compared with the control condition. The data suggest that the OGD has general suppressive effect on the gene expression with the exception of some genes which are related with ischemic cell death directly or indirectly. These genes are mainly involved in apoptotic and protein translation pathways and gap junction component. These results suggest that microarray analysis of gene expression may be useful for screening novel molecular mediators of astrocyte response to ischemic injury and making profound understanding of the cellular mechanisms as a whole. Such a screening technique should provide insights into the molecular basis of brain disorders and help to identify potential targets for therapy.

• 한국식품과학회지
• /
• 제45권5호
• /
• pp.590-597
• /
• 2013

고전압 펄스 전기장 기술은 가열 공정을 대체할 수 있는 비가열 살균기술로서 꾸준한 연구가 이루어지고 있는 기술이다. 본 실험에서는 미생물의 생리적 측면에서 고전압 펄스 전기장 처리에 의한 효모의 생리적 변화를 살펴보았다. 내염성의 변화를 측정한 결과 사멸하지 않은 세포의 90-99% 이상이 세포 손상에 의해 내염성을 소실하였으며, 내염성을 회복하는데 약 4-6시간의 시간이 걸렸다. 고전압 펄스 전기장 처리 초기에는 세포 내외의 pH 구배(${\Delta}pH$)가 차이가 컸으나 처리시간이 증가함에 따라 그 차이가 감소하였다. 이는 세포의 pH 항상성을 유지시키는 $H^+$-traslocation 기능을 담당하고 있는 $H^+$-ATPase 활성이 소실되었거나 세포막 자체가 손상되어 pH 항상성을 잃어버렸기 때문으로, 이는 고전압 펄스 전기장 처리한 세포의 $H^+$-ATPase의 활성이 초기부터 크게 감소하는 것으로 확인할 수 있었다. 또한 세포의 해당활성이 고전압 펄스 전기장 처리에 의해 크게 감소하여 세포의 대사 기능이 손상되는 것을 확인하였다.

• 한국근적외분광분석학회:학술대회논문집
• /
• 한국근적외분광분석학회 2001년도 NIR-2001
• /
• pp.4105-4105
• /
• 2001

Near infrared spectroscopy (NIRS) was employed to qualify and quantify on survival, the injury rate and apoptosis of the human breast cancer cell line MCF-7 cells. MCF-7 cells were cultured in RPMI medium supplemented with 10% FCS in a 95% air and 5% CO2 atmosphere at 37$^{\circ}C$. For the viable cells preparation, cells were de-touched by 0.1% of trypsin treatment and washed with RPMI supplemented with 10% FCS medium by centrifugation at 1000 rpm for 3min. For the dead cells preparation, cells were de-touched by a cell scraper. The cells were counted by a hemacytometer, and the viability was estimated by the exclusion method with frypan blue dye. Each viable and dead cells were suspended in PBS (phosphate bufferred saline) or milk at the cell density desired. For the quantitative determination of cell death by measuring the LDH (lactate dehydrogenase) activity liberated from cells with cell membrane injuries, LDH-Cytotoxic Test Wako (Wako, Pure Pharmaceutical Co. Ltd., Japan) was used. We found that NIRS measurement of MCF-7 cells at the density range could evaluate and monitor the different characteristics of living cells and dead cells. The spectral analysis was performed in two wavelength ranges and with 1,4, 10 mm pathlength. Different spectral data pretreatment and chemometrics methods were used. We applied SIMCA classificator on spectral data of living and dead cells and obtained good accuracy when identifying each class. Bigger variation in the spectra of living cells with different concentrations was observed when compared to the same concentrations of dead cells. PLS was used to measure the number of cells in PBS. The best model for measurement of dead cells, as well as living cells, was developed when raw spectra in the 600-1098 nm region and 4 mm pathlength were used. Smoothing and second derivative spectral data pretreatment gave worst results. The analysis of PLS loading explained this result with the scatter effect found in the raw spectra and increased with the number of cells. Calibration for cell count in the 1100-2500 nm region showed to be very inaccurate.

• Biomolecules & Therapeutics
• /
• 제32권5호
• /
• pp.647-657
• /
• 2024

Gefitinib is the well-tolerated first-line treatment of non-small cell lung cancer. As it needs analgesics during oncology treatment, particularly in the context of the coronavirus disease, where patients are more susceptible to contract high fever and sore throat. This has increased the likelihood of taking both gefitinib and antipyretic analgesic acetaminophen (APAP). Given that gefitinib and APAP overdose can predispose patients to liver injury or even acute liver failure, there is a risk of severe hepatotoxicity when these two drugs are used concomitantly. However, little is known regarding their safety at therapeutic doses. This study simulated the administration of gefitinib and APAP at clinically relevant doses in an animal model and confirmed that gefitinib in combination with APAP exhibited additional hepatotoxicity. We found that gefitinib plus APAP significantly exacerbated cell death, whereas each drug by itself had little or minor effect on hepatocyte survival. Mechanistically, combination of gefitinib and APAP induces hepatocyte death via the apoptotic pathway obviously. Reactive oxygen species (ROS) generation and DNA damage accumulation are involved in hepatocyte apoptosis. Gefitinib plus APAP also promotes the expression of Kelch-like ECH-associated protein 1 (Keap1) and downregulated the antioxidant factor, Nuclear factor erythroid 2-related factor 2 (Nrf2), by inhibiting p62 expression. Taken together, this study revealed the potential ROS-mediated apoptosis-dependent hepatotoxicity effect of the combination of gefitinib and APAP, in which the p62/Keap1/Nrf2 signaling pathway participates and plays an important regulatory role.

• 한국식품영양과학회지
• /
• 제44권1호
• /
• pp.35-43
• /
• 2015

본 연구에서는 노루궁뎅이 버섯균사체를 비타민나무 잎에 배양하여 조제한 노루궁뎅이 버섯균사체-비타민나무 발효물 열수 추출물이 생체고분자의 산화적 손상과 세포사멸을 보호할 수 있는지를 관찰하였다. 노루궁뎅이 버섯균사체-비타민나무 발효물의 항산화 활성을 DPPH radical, ABTS radical, peroxyl radical 소거활성 측정을 통해 알아보았다. 그 결과 노루궁뎅이 버섯균사체-비타민나무 발효물을 처리한 DPPH radical 소거활성은 $500{\mu}g/mL$ 농도에서 65.06%, ABTS radical 소거활성은 $50{\mu}g/mL$ 농도에서 98.83%, peroxyl radical 소거활성은 $100{\mu}g/mL$ 농도에서 44.03%로 높은 항산화 활성을 나타내었다. 노루궁뎅이 버섯균사체-비타민나무 발효물은 DNA의 산화적 손상을 효과적으로 억제하였다. 노루궁뎅이 버섯균사체-비타민나무 발효물 역시 사람의 혈청단백질과 Cu,Zn-SOD의 산화적 손상을 억제하였다. 세포에 $H_2O_2$를 처리하였을 때 세포생존율에 비하여 발효물을 $100{\mu}g/mL$ 농도로 전 처리한 세포생존율은 21.59% 높게 증가되었다. 또한 발효물을 $50{\mu}g/mL$ 농도로 처리했을 경우 세포 내 ROS의 축적이 유의적으로 감소되었다. 따라서 노루궁뎅이 버섯균사체-비타민나무 발효물은 항산화 활성뿐만 아니라 산화적 스트레스에 의해 야기되는 세포 독성에 대한 보호 작용이 뛰어난 것으로 사료된다.

• 대한임상검사과학회지
• /
• 제51권3호
• /
• pp.370-378
• /
• 2019

허혈 재관류 손상은 기관 이식, 외과적 혈관 재개통 및 출혈시에 발생하며 조직 및 기관 기능 장애를 유발한다. 최근 허혈 재관류 손상의 기전적 연구를 위해 간장 허혈 모델을 많이 이용하고 있다. 본 연구는 허혈 재관류 랫드 모델을 이용하여 항산화와 항염증 효과를 가진 것으로 알려진 Vanillin에 의한 간장 및 신장 손상에 대한 보호 효과를 알아보고 관련된 기전을 조사하기 위하여 실시하였다. 시험물질은 각각 100 mg/kg의 농도로 3일간 투여한 후, 60분동안 간을 결찰하여 허혈 재관류를 유도하여 관찰하였으며, 음성대조군, sham대조군 및 허혈재관류만 실시한 허혈 재관류대조군을 따로 두어, 약물투여군과 비교하였다. Vanillin 처치군에서는 AST, ALT 활성이 허혈 재관류대조군에 비해 유의하게 억제되었고, 조직병리학적 관찰에서도 염증 부분과 괴사부분이 현저하게 감소하였다. MDA와 SOD는 허혈 재관류군에 비해 유의적인 변화를 보였다. 이상의 결과를 종합하면 Vanillin은 간장 허혈 재관류에 의한 세포염증 및 세포괴사를 완화시켜 간세포 보호작용을 나타내었고, 신장의 사구체 및 원위세뇨관에 염증 변화를 완화 시키고 있어 세포 손상을 방어하는 것으로 생각되며, 이러한 방어효과는 항산화 기능에 의한 영향으로 사료된다.

• Journal of Chest Surgery
• /
• 제49권4호
• /
• pp.232-241
• /
• 2016

Background: Paraplegia is a devastating complication following operations on the thoracoabdominal aorta. We investigated whether histidine-tryptophan-ketoglutarate (HTK) solution could reduce the extent of ischemia/reperfusion (IR) spinal cord injuries in a rat model using a direct delivery method. Methods: Twenty-four Sprague-Dawley male rats were randomly divided into four groups. The sham group (n=6) underwent a sham operation, the IR group (n=6) underwent only an aortic occlusion, the saline infusion group (saline group, n=6) underwent an aortic occlusion and direct infusion of cold saline into the occluded aortic segment, and the HTK infusion group (HTK group, n=6) underwent an aortic occlusion and direct infusion of cold HTK solution into the occluded aortic segment. An IR spinal cord injury was induced by transabdominal clamping of the aorta distally to the left renal artery and proximally to the aortic bifurcation for 60 minutes. A neurological evaluation of locomotor function was performed using the modified Tarlov score after 48 hours of reperfusion. The spinal cord was harvested for histopathological and immunohistochemical examinations. Results: The spinal cord IR model using direct drug delivery in rats was highly reproducible. The Tarlov score was 4.0 in the sham group, $1.17{\pm}0.75$ in the IR group, $1.33{\pm}1.03$ in the saline group, and $2.67{\pm}0.81$ in the HTK group (p=0.04). The histopathological analysis of the HTK group showed reduced neuronal cell death. Conclusion: Direct infusion of cold HTK solution into the occluded aortic segment may reduce the extent of spinal cord injuries in an IR model in rats.

• 한방재활의학과학회지
• /
• 제20권4호
• /
• pp.1-15
• /
• 2010

Objectives : This study was performed to evaluate the effects of Hwangryunhaedok-tang(Huanglianjiedu-tang HHT) water extract on locomotor dysfunction induced by spinal cord injury(SCI) in rats. Methods : SCI was induced by mechanical contusion following laminectomy of 10th thoracic vertebra in Sprague-Dawley rats. HHT was orally given once a day for 14 days after SCI. Neurological behavior was examined with the Basso-Beattie-Bresnahan locomotor rating scale. Tissue damage and nerve fiber degeneration were examined with cresyl violet and luxol fast blue staining. Using immunohistochemisty, cellular damage to neurons and nerve fibers were examined against Bax and MAP-2. As inflammatory response markers, iNOS and COX-2 expressions were also examined. Results : 1. HHT ameliorated the locomotor dysfunction of the SCI-induced rats. 2. HHT attenuated the reduction of motor neurons in the ventral horn of the SCI-induced rat spinal cord. 3. HHT significantly reduced the number of Bax positive cells in the peri-lesion of the SCI-induced rat spinal cord. 4. HHT attenuated the reduction of MAP-2 positive cells in the peri-lesion of the SCI-induced rat spinal cord. 5. HHT significantly reduced the number of iNOS and COX-2 positive cells in the peri-lesion of the SCI-induced rat spinal cord. Conclusions : These results suggest that HHT improves the locomotor dysfunction of SCI by protecting motor neurons from cell death through anti-inflammatory effect.