• Nutrition Research and Practice
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• 제12권3호
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• pp.191-198
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• 2018

BACKGROUD/OBJECTIVES: Neuroinflammation plays critical role in neurodegenerative disorders, such as Alzheimer's disease (AD). We investigated the effect of three licorice varieties, Glycyrhiza uralensis, G. glabra, and Shinwongam (SW) on a mouse model of inflammation-induced memory and cognitive deficit. MATERIALS/METHODS: C57BL/6 mice were injected with lipopolysaccharide (LPS; 2.5 mg/kg, intraperitoneally) and orally administrated G. uralensis, G. glabra, and SW extract (150 mg/kg/day). SW, a new species of licorice in Korea, was combined with G. uralensis and G. glabra. Behavioral tests, including the T-maze, novel object recognition and Morris water maze, were carried out to assess learning and memory. In addition, the expressions of inflammation-related proteins in brain tissue were measured by western blotting. RESULTS: There was a significant decrease in spatial and objective recognition memory in LPS-induced cognitive impairment group, as measured by the T-maze and novel object recognition test; however, the administration of licorice ameliorated these deficits. In addition, licorice-treated groups exhibited improved learning and memory ability in the Morris water maze. Furthermore, LPS-injected mice had up-regulated pro-inflammatory proteins, such as inducible nitric oxide synthase (iNOS), cyclooxygenase-2, interleukin-6, via activation of toll like receptor 4 (TLR4) and nuclear factor-kappa B ($NF{\kappa}B$) pathways in the brain. However, these were attenuated by following administration of the three licorice varieties. Interestingly, the SW-administered group showed greater inhibition of iNOS and TLR4 when compared with the other licorice varieties. Furthermore, there was a significant increase in the expression of brain-derived neurotrophic factor (BDNF) in the brain of LPS-induced cognitively impaired mice that were administered licorice, with the greatest effect following SW treatment. CONCLUSIONS: The three licorice varieties ameliorated the inflammation-induced cognitive dysfunction by down-regulating inflammatory proteins and up-regulating BDNF. These results suggest that licorice, in particular SW, could be potential therapeutic agents against cognitive impairment.

• Journal of Ginseng Research
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• 제45권6호
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• pp.665-675
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• 2021

Background: Ginsenoside Rg1 (Rg1), an active ingredient in ginseng, may be a potential agent for the treatment of Alzheimer's disease (AD). However, the protective effect of Rg1 on neurodegeneration in AD and its mechanism of action are still incompletely understood. Methods: Wild type (WT) and APP/PS1 AD mice, from 6 to 9 months old, were used in the experiment. The open field test (OFT) and Morris water maze (MWM) were used to detect behavioral changes. Neuronal damage was assessed by hematoxylin and eosin (H&E) and Nissl staining. Immunofluorescence, western blotting, and quantitative real-time polymerase chain reaction (q-PCR) were used to examine postsynaptic density 95 (PSD95) expression, amyloid beta (Aβ) deposition, Tau and phosphorylated Tau (p-Tau) expression, reactive oxygen species (ROS) production, and NAPDH oxidase 2 (NOX2) expression. Results: Rg1 treatment for 12 weeks significantly ameliorated cognitive impairments and neuronal damage and decreased the p-Tau level, amyloid precursor protein (APP) expression, and Aβ generation in APP/PS1 mice. Meanwhile, Rg1 treatment significantly decreased the ROS level and NOX2 expression in the hippocampus and cortex of APP/PS1 mice. Conclusions: Rg1 alleviates cognitive impairments, neuronal damage, and reduce Aβ deposition by inhibiting NOX2 activation in APP/PS1 mice.

• 운동영양학회지
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• 제13권1호
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• pp.1-7
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• 2009

It has been shown that both caloric restriction and exercise, enhances glucose uptake through translocation of GLUT-4 protein. It remains unclear how exercise and caloric restriction affect the changes in VAMP (vesicle-associated membrane protein) in skeletal muscle and GLUT-2 in liver. This study investigated the effects of exercise training and caloric restriction on the expressions of glucose transport relating proteins in muscle and liver tissues in diabetic rats. Forty male Sprague-Dawley rats (250±10 g; 8 week in age) were assigned equally to four different groups; control (C), exercise only (E), dietary restriction only (D) and dietary restriction and exercise (DE). Daily food consumption was monitored to establish baseline intake. Both C and E groups consumed baseline food intake while D and DE groups were provided with only 60% of baseline total food intake. Forty-eight hours after intraperitoneal injection of STZ (50 mg/kg), diabetes was confirmed (8-hr fasting blood glucose levels ≥300 mg/dl). Rats in the E and DE groups exercised on a motorized treadmill for 30 min/d, 5 days/week for 4 weeks (5 min running at 3 m/min, 0% grade; 8 m/min for the next 5min, and then 15 m/min for 20 min). Rats were sacrificed 48 hrs after the last bout of exercise. Soleus muscle and liver were extracted to analyze for GLUT-4, VAMP-2, and GLUT-2, respectively. All variables were analyzed using the Western Blotting technique. All values were expressed as optical volume measured by optical density. A Two-way ANOVA was used to examine the difference between groups and applied Duncan's test for post-hoc. No significant differences in GLUT-2 expression were found among groups. However, E (280133±13228 arbitrary units{AU}) and DE (268833±14424 AU) groups showed significantly higher (p<.001) levels of GLUT-4 as compared with C (34461±2099 AU) and D groups (27847±703 AU). VAMP-2 protein expression increased (p<.001) in E (184137±7803 AU) and DE (189800±10856 AU) groups as compared to C (74201±8296AU) and D (72967±863 AU) groups. These results suggest that either exercise with or without caloric restriction increases the up-regulation of GLUT-4 and VAMP-2 in skeletal muscle of diabetic rats. However, GLUT-2 protein in liver was not affected by either exercise or exercise with caloric restriction.

• Korean Journal of Acupuncture
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• 제41권2호
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• pp.33-42
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• 2024

Objectives : The sigma-1 receptor is implicated in stress, depression, psychostimulant sensitization, and addiction vulnerability. Prior studies have indicated that ethanol exposure modulates sigma-1 receptor activity within the Ventral Tegmental Area (VTA). Here, we explore the sub-mechanisms underlying sigma-1 receptor activity induced by HT7 (Shinmun) stimulation in behavioral alterations following acute ethanol (ETOH) administration. Methods : Male Wistar rats were investigated for pro- and anti-inflammatory markers after injection of ETOH (1 g/kg) using cytokine enzyme-linked immunosorbent assay (ELISA)s. After confirming that HT7 stimulation changed the total distance traveled in the open field test (OFT), protein changes in the Ventral tegmental area (VTA) were measured by Western blotting. The expression level of inducible nitric oxide synthase (iNOS) after administration of a sigma-1 receptor antagonist (dihydrobromide 1047; BD1047, 10 mg/kg i.p.) and Shenmen (HT7) stimulation was compared. Results : As a result, acute ETOH administration increased proinflammatory marker levels (TNF-𝛼 and IL-6). HT7 stimulation restored the total distance response after acute ethanol administration. In addition, in the VTA, the levels of a microglial marker (iNOS), sigma-1 receptor and protein kinase C, which are predicted to be involved in up- and downregulation, were restored by HT7 stimulation. In particular, HT7 stimulation modulates iNOS expression through effects similar to BD treatment. This study suggests that the stimulatory effect of HT7 may be driven by microglial activation. Conclusions : Microglial activity is regulated by sigma-1 receptor, and sigma-1 receptor activity is regulated by HT7 stimulation. Significantly, we demonstrate that HT7 stimulation ameliorates behavioral alterations induced by acute ETOH administration through microglial activation within the VTA.

• Asian Pacific Journal of Cancer Prevention
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• 제15권12호
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• pp.4773-4780
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• 2014

Background: To investigate the effects of small interference RNA (siRNA) targeting BCR/ABL mRNA on proliferation and apoptosis in the K562 human chronic myeloid leukemia (CML) cell line and to provide a theoretical rationale and experimental evidence for its potential clinical application for anti-CML treatment. Materials and Methods: The gene sequence for BCR/ABL mRNA was found from the GeneBank. The target gene site on the BCR/ABL mRNA were selected according to Max-Planck-Institute (MPI) and rational siRNA design rules, the secondary structure of the candidate targeted mRNA was predicted, the relevant thermodynamic parameters were analyzed, and the targeted gene sequences were compared with BLAST to eliminate any sequences with significant homology. Inhibition of proliferation was evaluated by MTT assay and colony-formation inhibiting test. Apoptosis was determined by flow cytometry (FCM) and the morphology of apoptotic cells was identified by Giemsa-Wright staining. Western blotting was used to analyze the expression of BCR/ABL fusion protein in K562 cells after siRNA treatment. Results: The mRNA local secondary structure calculated by RNA structure software, and the optimal design of specific siRNA were contributed by bioinformatics rules. Five sequences of BCR/ABL siRNAs were designed and synthesized in vitro. Three sequences, siRNA1384, siRNA1276 and siRNA1786, which showed the most effective inhibition of K562 cell growth, were identified among the five candidate siRNAs, with a cell proliferative inhibitory rate nearly 50% after exposure to 12.5nmol/L~50nmol/L siRNA1384 for 24,48 and 72 hours. The 50% inhibitory concentrations ($IC_{50}$) of siRNA1384, siRNA1276 and siRNA1786 for 24hours were 46.6 nmol/L, 59.3 nmol/L and 62.6 nmol/L, respectively, and 65.668 nmol/L, 76.6 nmol/L, 74.4 nmol/L for 72 hours. The colony-formation inhibiting test also indicated that, compared with control, cell growth of siRNA treated group was inhibited. FCM results showed that the rate of cell apoptosis increased 24 hours after transfecting siRNA. The results of annexinV/PI staining indicated that the rate of apoptosis imcreased (1.53%, 15.3%, 64.5%, 57.5% and 21.5%) following treamtne with siRNAs (siRNA34, siRNA372, siRNA1384, siRNA1276 and siRNA1786). Morphological analysis showed td typical morphologic changes of apoptosis such as shrunken, fragmentation nucleus as well as "apoptotic bodies" after K562 cell exposure to siRNA. Western blot analysis showed that BCR/ABL protein was reduced sharply after a single dose of 50nmol/L siRNA transfection. Conclusions: Proliferation of K562 cells was remarkbly inhibited by siRNAs (siRNA1384, siRNA1276 and siRNA1786) in a concentration-dependent manner in vitro, with effective induction of apoptosis at a concentration of 50 nmol/L. One anti-leukemia mechanism in K562 cells appeared that BCR/ABL targeted protein was highly down-regulated. The siRNAs (siRNA1384, siRNA1276 and siRNA1786) may prove valuable in the treatment of CML.

• Journal of Ginseng Research
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• 제44권3호
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• pp.442-452
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• 2020

Backgroud: Sleep deprivation (SD) impairs learning and memory by inhibiting hippocampal functioning at molecular and cellular levels. Abnormal autophagy and apoptosis are closely associated with neurodegeneration in the central nervous system. This study is aimed to explore the alleviative effect and the underlying molecular mechanism of stem-leaf saponins of Panax notoginseng (SLSP) on the abnormal neuronal autophagy and apoptosis in hippocampus of mice with impaired learning and memory induced by SD. Methods: Mouse spatial learning and memory were assessed by Morris water maze test. Neuronal morphological changes were observed by Nissl staining. Autophagosome formation was examined by transmission electron microscopy, immunofluorescent staining, acridine orange staining, and transient transfection of the tf-LC3 plasmid. Apoptotic event was analyzed by flow cytometry after PI/annexin V staining. The expression or activation of autophagy and apoptosis-related proteins were detected by Western blotting assay. Results: SLSP was shown to improve the spatial learning and memory of mice after SD for 48 h, accomanied with restrained excessive autophage and apoptosis, whereas enhanced activation of phosphoinositide 3-kinase/protein kinase B/mammalian target of rapamycin signaling pathway in hippocampal neurons. Meanwhile, it improved the aberrant autophagy and apoptosis induced by rapamycin and re-activated phosphoinositide 3-kinase/Akt/mammalian target of rapamycin signaling transduction in HT-22 cells, a hippocampal neuronal cell line. Conclusion: SLSP could alleviate cognitive impairment induced by SD, which was achieved probably through suppressing the abnormal autophagy and apoptosis of hippocampal neurons. The findings may contribute to the clinical application of SLSP in the prevention or therapy of neurological disorders associated with SD.

• 한국식품영양과학회지
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• 제45권8호
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• pp.1090-1098
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• 2016

본 연구는 해조류 중에서도 홍조류인 주름까막살 에탄올 추출물(GCEE)의 항염증 효과를 확인하기 위한 실험으로 proinflammatory cytokines의 분비량 및 iNOS, COX-2, $NF-{\kappa}B$와 MAPKs의 발현량을 관찰하고 마우스모델에서 항염증 효과를 확인하였다. GCEE가 세포 생존율에 있어 독성을 나타내지 않음을 MTT assay를 통해 확인한 후 같은 농도로 추후실험을 진행하였다. NO 분비량이 GCEE에 의해 농도 의존적으로 감소하였으며 전염증성 매개물질인 사이토카인(IL-6, $TNF-{\alpha}$ 및 $IL-1{\beta}$)의 분비량 또한 유의적으로 감소하였다. 이러한 결과가 전염증성 매개인자의 전사인자인 $NF-{\kappa}B$와 MAPKs 경로에 의한 것인지 확인하기 위하여 발현량을 관찰한 결과, GCEE가 LPS 처리로 현저히 증가한 염증 관련 효소인 iNOS와 COX-2의 단백질 발현을 농도 의존적으로 유의성 있게 억제하였고 전사인자인 $NF-{\kappa}B$ 및 MAPKs의 발현을 억제하였다. 또한, GCEE는 croton oil로 부종을 유발한 마우스모델에서 귀 부종 억제 효과를 나타내었고 250 mg/kg 농도에서 조직의 경피 및 진피 두께의 발달을 prednisolone 50 mg/kg 처리구와 유사한 정도까지 현저히 억제하고 염증성 세포인 mast cell의 침윤 억제 효과도 확인하였다. 이를 통해 주름까막살 에탄올 추출물은 염증반응의 전사인자인 $NF-{\kappa}B$와 MAPKs의 발현을 조절함으로써 iNOS와 COX-2의 발현을 억제하고 그에 따라 전염증성 매 개인자인 NO, IL-6, $TNF-{\alpha}$ 및 $IL-1{\beta}$의 분비를 억제하여 항염증 활성을 가지는 것을 확인하였으며, 이 결과를 종합해 볼 때 주름까막살 에탄올 추출물이 염증 치료제의 소재로 이용될 가치가 충분할 것으로 생각한다.

• Parasites, Hosts and Diseases
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• 제29권4호
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• pp.339-354
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• 1991

스파르가눔을 흰쥐와 흰생쥐에게 인긍 구강감염을 시켜 감염 2주후부터 숙주의 기생부위에서 충체와 함께 둘레의 조직을 적출하여 동결절편을 만들고 이것을 ABC면역효소표지법과 간접형광항체법 (IFA)으로 항원성분의 분포와 그 추이를 추구한 결과, IgG 항체에 의한 면역반응이 충체의 망상유조직에서 인지되었으며 특히 ABC면역효소표지법에 의하면 유조직 중 수질에서 보다 피질층에서 강한 반응성을 나타났다. 이 중에서도 석회소체가 있는 곳에 염색 반응성이 강하였다. 감염후기에서는 스파르가눔의 외피 (tegument) 내외면과 충체를 둘러 싸고 있는 피낭의 막면, 피막 밖에 있는 섬유성 결합조직과 근조직의 간극에서도 항원성분을 관찰할 수 있다. 특히 결합조직 사이에 분포된 소혈관의 둘레와 현관 외벽에 강한 반응이 나타났다. IgM 항체에 인지된 항원성분도 충체의 유조직층중 피질충에서 강한 반응이 나타났다. 스파르가눔의 추출 조항원성분과 배설분비 항원성분을 SDS-PAGE한 후 nitrocellulose 막에 영동전이하여 IgG, IgM 항체를 유도하는 항원성분을 ABC효소표지 항체 법으로 조사하였다. 스파르가눔의 추출 조항원 성분 중 흰쥐의 IgG 항체에 인지된 항원 성분은 23개이고 IsM 항체에는 15개의 항원성분이 인지되었다. 흰생쥐에서는 IgG 항체로 I6개 항원성분이, IgM 항체에 9개의 항원성분이 인지되었다. 배설분비 항원성분 중 횐쥐의 IgG 항체에 20개 항원성분이, IgM 항체에는 5개의 항원 성분이 인지되었다. 횐생쥐의 IgG 항체에 18개, IgM 항체에 11개의 항원성분이 인지되었다. 조창원성분 중 15개 항원성분과 배설분비 항원성분중 13개의 항원 성분이 흰쥐, 흰생쥐의 IgG 항체에 모두 교차반응이 일어났으며 IgM 항체에 반응한 항원성분중 IgG 항체와도 교차반응이 있었다.

• 한국식품저장유통학회지
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• 제25권1호
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• pp.107-116
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• 2018

본 연구의 목적은 블루베리 잎의 용매별 추출물에 따른 약리활성에 대한 검증 및 효능 평가이다. 블루베리 잎의 항산화, 항염증에 대한 효과를 확인하였다. 염증 반응은 자극이 가해지면 histamine, serotonin, prostaglandin과 같은 혈관 활성물질에 의해 혈관 투과성이 증대되어 염증을 유발하고 cytokine, free radical, lysosomal enzyme 등 다양한 매개 인자가 관여한다. 자극에 의한 macrophage cell의 염증반응은 tumor necrosis factor-${\alpha}$(TNF-${\alpha}$), interleukin-6(IL-6), interleukin-$1{\beta}$(IL-$1{\beta}$)와 같은 pro-inflammatory cytokine의 발현이 유도되고, inducible nitric oxide synthase(i-NOS)와 cyclooxygenase-2(COX-2)에 영향을 받는 유전자의 발현을 자극하게 되어 nitric oxide(NO) 및 $PGE_2$등의 염증 인자가 생성된다. 이에 따라 블루베리 잎 추출물의 항염증에 대한 연구를 위해 이에 영향을 주는 인자인 i-NOS, COX-2의 단백질 발현억제 작용을 확인 하였다. 그 결과 BLA > BLE > BLW 순서로 높은 효능을 확인 할 수 있었다. 가장 효과가 좋은 BLA 처리군에서 $PGE_2$ 분비량 및 다양한 염증성 인자의 mRNA 발현량을 확인하였다. 측정 결과, BLA($100{\mu}g/mL$)는 $PGE_2$ 분비량을 85.3% 억제하였으며 i-NOS, COX-2, IL-$1{\beta}$, IL-6, TNF-${\alpha}$ 단백질과 mRNA 발현이 각각 86.8%, 85.7%, 62.7%, 77%, 32.2% 억제되는 효과를 확인할 수 있었다. 결론적으로 블루베리 70% 아세톤 추출물(BLA)의 항염증 효과가 가장 높았으며, 블루베리 잎은 세포의 mRNA 및 단백질 수준에서 염증인자들의 억제를 통해 대식세포에서 항염증 효과가 명백히 확인되었다. 더 나아가, nuclear factor kappa B(NF-${\kappa}B$), signal transducer and activator of transcription(STAT-1), mitogen-activated protein kinases(MAPKs) 등의 세포 내 염증관련 중간기전을 연구해볼 가치가 있다고 사료된다.