• Journal of Acupuncture Research
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• 제29권1호
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• pp.37-45
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• 2012

목적 : 최근 NF-${\kappa}B$와 STAT3의 활성억제와 관련된 항암제 연구가 주목받고 있으며, 본 연구는 사독(蛇毒)이 세포자멸사 관련 단백질의 발현 조절을 통하여 세포자멸사를 유도하고, NF-${\kappa}B$와 STAT3의 활성억제를 유도하여 난소암 PA-1 세포의 성장을 억제하는지를 확인하고, 해당 기전을 살펴보고자 하였다. 방법 : 사독을 처리한 후 난소암 PA-1 세포의 세포자멸사의 관찰에는 DAPI, TUNEL staining assay를 시행하였고, 세포자멸사 조절단백질 및 NF-${\kappa}B$, STAT3의 활성 변동 관찰에는 western blot analysis를 시행하였다. 결과 : 1. 사독을 처리한 후 난소암 PA-1 세포에서 세포자멸사가 유도되어 암세포성장이 억제되었다. 2. 사독을 처리한 후 세포자멸사 관련 단백질 중 세포자멸사 촉진 단백질인 cleaved caspase-3, Bax의 발현은 증가되었고, 세포자멸사 억제 단백질인 Bcl-2의 발현은 감소되었다. 3. 사독을 처리한 후 난소암 PA-1 세포의 NF-${\kappa}B$와 STAT3 발현은 감소되었고, 각각의 길항제인 salicylic acid와 stattic 처리 후 NF-${\kappa}B$와 STAT3 발현은 더욱 감소되었다. 결론 : 사독은 난소암 세포의 세포자멸사 유발과, NF-${\kappa}B$와 STAT3의 활성억제를 통해 치료 효율이 높고, 내성이 적은 난소암 치료제의 개발에 도움이 될 것으로 기대된다.

• International Journal of Oral Biology
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• 제33권2호
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• pp.59-64
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• 2008

This study was designed to investigate the changes in the properties of the neuronal setm cells or progenitor cells associated with age-related decline in neurogenesis of the hippocampal dentate gyrus (DG). Active whole cells cycle marker Ki67 (a marker of whole cell cycle)-positive and S phase marker bromodeoxyuridine (BrdU)-positive. Neural stem cells gradually were reduced in the hippocampal subgranular zone (SGZ) in an age-dependant manner after birth (from P1 month to P1 year). The ratio of BrdUpositivecells/Ki67-positive cells was gradually enhanced in an age-dependent manner. The ratio of Ki67-positive cells/accu-mulating BrdU-positive cells at 3 hrs after BrdU injection was injected once a day for consecutive 5 days gradually decreased during ageing. TUNEL- and caspase 3 (apoptotic terminal caspase)-positive cells gradually decreased in the dentate SGZ during ageing and immunohistochemical findings of glial fibrillary acid protein (GFAP) were not changed during ageing. NeuN, a marker of mature neural cells, and BrdU-double positive cells gradually decreased in an age-dependent manner but differentiating ratio and survival rate of cells were not changed at 4 wks after BrdU injection once a day for consecutive 5 days. The number of BrdU-positive cells migrated from the hippocampal SGZ into granular layer and its migration speed was gradually declined during ageing. These results suggest that the adult neurogenesis in the mouse hippocampal DG gradually decrease through reducing proliferation of neural stem cells accompanying with cells cycle change and reduced cells migration rather than changes of differentiation.

• Biomolecules & Therapeutics
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• 제18권1호
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• pp.48-55
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• 2010

Neural progenitor cells (NPCs) differentiate into astrocytes, neurons and oligodendrocytes, which is controlled by various factors in brain. Recent evidences suggest that small molecules modulating the proliferation and differentiation of NPCs may have therapeutic value as well as the potential use as chemical probes. Phylligenin is a lignan with anti-inflammatory activity that is isolated from the fruits of Forsythia koreana. We investigated effects of phylligenin on proliferation and differentiation of NPCs. Treatment of phylligenin decreased the number of proliferating NPCs in culture without effects on the differentiation and survival of neural cells such as neurons and astrocytes. To examine the mechanism of the decreased NPCs number, we performed cell cycle analysis. Proliferation of NPCs was decreased via G1-S transition block by phylligenin treatment, and it was mediated by the increase of p21 level. However, phylligenin did not induce apoptosis of NPCs as determined by TUNEL assay and PARP cleavage. We also found that viability of glioma cell lines such as C6 and U87MG glioma cells, but not that of primary neuron and astrocyte, was inhibited by phylligenin. These results suggest that phylligenin selectively inhibits proliferation of rapidly growing cells such as neural stem cells and glioma cells. Given that the possible role of brain tumor stem cells in the pathology of brain cancers, the inhibitory effects of phylligenin might be useful in the development of new therapeutic agents against brain cancers.

• Biomolecules & Therapeutics
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• 제21권1호
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• pp.84-88
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• 2013

High risk of cardiovascular diseases caused by existing PPAR-${\gamma}$ agonists such as rosiglitazone and pioglitazone has been recently reported. CKD-501 is a novel selective PPAR-${\gamma}$ agonist as a potential target to reduce cardiovascular risk in non-insulin dependent diabetes mellitus (NIDDM). In this study, We investigated potential cardiotoxicity of CKD-501 and compared its toxicity with that of rosiglitazone or pioglitazone using db/db mice. After 12-week repeated administration of CKD-501 at doses of 3, 10 and 30 mg/kg/day or rosiglitazone at doses of 10 and 30 mg/kg/day or pioglitazone at doses of 200 and 540 mg/kg/day, animals were sacrificed for investigation of potential toxicities. Diameters of left ventricles and areas of cardiomyocytes were measured. And lipid accumulation and apoptosis in heart muscle were examined by oil red O staining and TUNEL staining, respectively. Diameters of left ventricles were significantly increased in high dose treatment group of pioglitazone compared to control (p<0.05), while other groups showed a tendency for an increase. All test articles induced significantly the increase of area of cardiomyocytes in heart compared to control (p<0.01), in regular order as pioglitazone > CKD-501 ${\geq}$ rosiglitazone. However, lipid accumulation and apoptotic changes in heart were not observed in all dosing groups. Taken together, the myocardial cell hypertrophy of CKD-501 are relatively lower than that of pioglitazone and similar to rosiglitazone. And it is suggested that the myocardial cell hypertrophy of CKD-501 are less adverse in clinical use for the management of the NIDDM.

• Restorative Dentistry and Endodontics
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• 제37권1호
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• pp.34-40
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• 2012

Objectives: The aim was to confirm the stem cell-like properties of the dental pulp stromal cells and to evaluate the morphologic changes during in vitro chondrogenesis. Materials and Methods: Stromal cells were outgrown from the dental pulp tissue of the premolars. Surface markers were investigated and cell proliferation rate was compared to other mesenchymal stem cells. Multipotency of the pulp cells was confirmed by inducing osteogenesis, adipogenesis and chondrogenesis. The morphologic changes in the chondrogenic pellet during the 21 day of induction were evaluated under light microscope and transmission electron microscope. TUNEL assay was used to evaluate apoptosis within the chondrogenic pellets. Results: Pulp cells were CD90, 105 positive and CD31, 34 negative. They showed similar proliferation rate to other stem cells. Pulp cells differentiated to osteogenic, adipogenic and chondrogenic tissues. During chondrogenesis, 3-dimensional pellet was created with multi-layers, hypertrophic chondrocyte-like cells and cartilage-like extracellular matrix. However, cell morphology became irregular and apoptotic cells were increased after 7 day of chondrogenic induction. Conclusions: Pulp cells indicated mesenchymal stem cell-like characteristics. During the in vitro chondrogenesis, cellular activity was superior during the earlier phase (within 7 day) of differentiation.

• KSBB Journal
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• 제31권1호
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• pp.52-57
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• 2016

Mutant p53 (R280K) is highly expressed in MDA-MB-231 triple-negative human breast cancer cells. Currently, we reported the role of mutant p53-R280K in mediating the survival of MDA-MB-231 cells in vitro. The present study was undertaken to determine whether mutant p53-R280K affects breast cancer cell growth in vivo. To this end, we used small interfering RNA to knockdown the level of mutant p53-R280K in MDA-MB-231 cells. Silencing of mutant p53-R280K in MDA-MB-231 cells causes substantial tumor regression of established xenografts in vivo. In xenograft model for breast cancer, silencing of mutant p53-R280K in MDA-MB-231 cells significantly inhibited the tumor growth. Moreover, TUNEL assay showed more occurrence of apoptotic cells in mutant p53-R280K silenced tumors compared to control. Our data indicate that mutant p53-R280K has an important role in mediating tumor growth of MDA-MB-231 cells in vivo. Taken together, this study suggests that endogenous mutant p53-R280K could be used as a therapeutic target for breast cancer cells harboring this TP53 missense mutation.

• The Korean Journal of Physiology and Pharmacology
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• 제24권5호
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• pp.423-431
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• 2020

This study aimed to evaluate the effect of curcumin on brain hypoxic-ischemic (HI) damage in neonatal rats and whether the phosphoinositide 3-kinase (PI3K)/Akt/vascular endothelial growth factor (VEGF) signaling pathway is involved. Brain HI damage models were established in neonatal rats, which received the following treatments: curcumin by intraperitoneal injection before injury, insulin-like growth factor 1 (IGF-1) by subcutaneous injection after injury, and VEGF by intracerebroventricular injection after injury. This was followed by neurological evaluation, hemodynamic measurements, histopathological assessment, TUNEL assay, flow cytometry, and western blotting to assess the expression of p-PI3K, PI3K, p-Akt, Akt, and VEGF. Compared with rats that underwent sham operation, rats with brain HI damage showed remarkably increased neurological deficits, reduced right blood flow volume, elevated blood viscosity and haematocrit, and aggravated cell damage and apoptosis; these injuries were significantly improved by curcumin pretreatment. Meanwhile, brain HI damage induced the overexpression of p-PI3K, p-Akt, and VEGF, while curcumin pretreatment inhibited the expression of these proteins. In addition, IGF-1 treatment rescued the curcumin-induced down-regulated expression of p-PI3K, p-Akt, and VEGF, and VEGF overexpression counteracted the inhibitory effect of curcumin on brain HI damage. Overall, pretreatment with curcumin protected against brain HI damage by targeting VEGF via the PI3K/Akt signaling pathway in neonatal rats.

• 한국동물위생학회지
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• 제42권3호
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• pp.135-144
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• 2019

Low pathogenic avian influenza (LPAI; H9N2) and Newcastle disease (ND) are economically important poultry diseases in Korea. In this study, we investigated pathological features and virus distribution in the lymphoid tissues of chicks experimentally infected with H9N2 and/or ND virus. Six-weeks-old SPF chickens were divided into 4 groups, Control (C), H9N2 (E1), NDV (E2), and H9N2+NDV (E3). E1 group was challenged with 0.1 ml A/Kr/Ck/01310/01 (H9N2) $10^{5.6}$ $EID_{50}$ intranasally, E2 group was challenged with 0.5 ml KJW (NDV) $10^{5.0}{\sim}10^{6.0}$ $ELD_{50}$ intramuscularly, and E3 group was challenged with H9N2, followed 7 days later by NDV. In histopathological examination, E1 group showed depletion and necrosis in bursa of Fabricius, thymus, cecal tonsil, and spleen, whereas E2 and E3 groups were noted severe lymphocyte depletion and necrosis with destruction of lymphoid organs structures. In TUNEL assay, apoptotic bodies were detected in lymphoid organs of all experimental groups, which was most severe in E3 group. H9N2 and ND viruses were predominantly detected in cecal tonsil of E1, E2, and E3 groups by PCR and immunohistochemistry (ICH). In conclusion, co-infection of H9N2 with NDV caused severe pathologic lesions and apoptosis in lymphoid tissues compared to single infections.

• 한국동물생명공학회지
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• 제35권4호
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• pp.347-356
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• 2020

Gangliosides are glycolipids in which oligosaccharide is combined with sialic acids. Our previous studies have suggested an interplay between ganglioside GD1a/GT1b and meiotic maturation capacity in porcine oocyte maturation. Furthermore, ganglioside GD1a and GT1b are known for its antioxidant activity, but it is still unclear whether possible antioxidant role of GD1a and GT1b is involved in porcine embryos development competence during in vitro culture (IVC). Here, the effects of ganglioside GD1a and GT1b on the embryonic developmental competence during in vitro culture of porcine were investigated. The effects of ganglioside GD1a and GT1b on the expression of ST3GAL2 were confirmed during embryos development (2-cell, 4-cell, 8-cell and blastocyst) using immunofluorescent staining (IF). As a result, the fluorescent expression of ST3GAl2 was higher in embryos at 4-8 cells stage than blastocysts. Blastocyst development rate significantly increased in only 0.1 μM GD1a and GT1b treated groups compared with control group. To investigate the cellular apoptosis, we analyzed TUNEL assay. In case of only 0.1 μM GD1a and GT1b treated groups, the total number of cells in blastocyst compared with control group, but there was no significant difference in the rate of apoptotic cells. We identified the intracellular ROS levels using DCF-DA staining. According to the result, ROS production significantly decreased in blastocysts derived from the 0.1 μM GD1a and GT1b treated groups. These results suggest that ganglioside GD1a and GT1b improve the developmental competence of porcine embryos via reduction of intracellular ROS during preimplantation stage.

• 운동영양학회지
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• 제13권2호
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• pp.115-122
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• 2009

Intracerebral hemorrhage (ICH) is a common cause of stroke, and it occurs mainly in the striatum, thalamus, cerebellum, and pons. Physical exercise is known to ameliorate neurologic impairment induced by various brain insults. In the present study, the influence of pre-and post-conditioning of treadmill exercise on spatial learning ability, the lesion volume, and apoptotic neuronal cell death in the striatum following ICH in rats was investigated. ICH in the striatum was induced by injection of collagenase using strereotaxic instrument. The rats in the pre-exercise group were scheduled to run on a treadmill before ICH induction for 2 consecutive weeks. The rats in the post-exercise group were scheduled to run on a treadmill after ICH induction for 2 weeks. The rats in the pre-exercise and post-exercise group were scheduled to run on a preconditioning treadmill exercise 2 weeks before ICH induction until postconditioning treadmill exercise 2 weeks after ICH induction, except the day of surgery. For this study, radial arm maze task, Nissl staining, terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay, and immunohistochemistry for caspase-3 were performed. Our date showed that treadmill exercise suppressed the ICH-induced apoptotic neuronal cell death and decreased lesion volume in the stratum. Treadmill exercise also alleviated the ICH-induced impairment of spatial learning ability. Preconditioning treadmill exercise before the ICH insult and postconditioning treadmill exercise after the ICH insult showed similar effectiveness on the recovery of ICH. In this study, however, preconditioning exercise before the ICH insult and postconditioning exercise after the ICH insult showed the most potent effectiveness on the recovery of ICH.