• Biomolecules & Therapeutics
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• 제27권5호
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• pp.492-501
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• 2019

Nitrogen-containing heterocycles such as quinoline, quinazolinones and indole are scaffolds of natural products and have broad biological effects. During the last years those structures have been intensively synthesized and modified to yield new synthetic molecules that can specifically inhibit the activity of dysregulated protein kinases in cancer cells. Herein, a series of newly synthesized isoquinolinamine (FX-1 to 8) and isoindoloquinazolinone (FX-9, FX-42, FX-43) compounds were evaluated in regards to their anti-leukemic potential on human B- and T- acute lymphoblastic leukemia (ALL) cells. Several biological effects were observed. B-ALL cells (SEM, RS4;11) were more sensitive against isoquinolinamine compounds than T-ALL cells (Jurkat, CEM). In SEM cells, metabolic activity decreased with $10{\mu}M$ up to 26.7% (FX-3), 25.2% (FX-7) and 14.5% (FX-8). The 3-(p-Tolyl) isoquinolin-1-amine FX-9 was the most effective agent against B- and T-ALL cells with IC50 values ranging from 0.54 to $1.94{\mu}M$. None of the tested compounds displayed hemolysis on erythrocytes or cytotoxicity against healthy leukocytes. Anti-proliferative effect of FX-9 was associated with changes in cell morphology and apoptosis induction. Further, influence of FX-9 on PI3K/AKT, MAPK and JAK/STAT signaling was detected but was heterogeneous. Functional inhibition testing of 58 kinases revealed no specific inhibitory activity among cancer-related kinases. In conclusion, FX-9 displays significant antileukemic activity in B- and T-ALL cells and should be further evaluated in regards to the mechanisms of action. Further compounds of the current series might serve as templates for the design of new compounds and as basic structures for modification approaches.

• 생명과학회지
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• 제31권3호
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• pp.266-279
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• 2021

본 연구에서는 다양한 사람의 암세포주(A-549, MCF-7, MDA-MB-231, U87-MG, AGS, MKN-74 및 SNU-601 세포)와 정상세포주(MRC-5 섬유아세포 및 사랑니 유래 중간엽성 줄기세포에 토복령 추출물(Smilax china L. extract, SCLE)을 처리하여 세포 사멸 효과를 조사하였다. SCLE 처리 후, MTT 분석에서 여러 암세포주는 정상세포주보다 유의적으로 휠씬 낮은 반억제농도값을 나타내었고, 세포는 세포부착력의 소실로 인한 세포사멸(anoikis)이 관찰되었다. 또한, SCLE를 처리한 A-549, AGS 및 MCF-7 암세포주에서 세포의 생존성과 말단소립 복원효소의 활성도를 조사하였을 때, SCLE 처리 후 4일째에 세포의 생존성과 말단소립 복원효소의 활성도가 현저히 줄어드는 것을 관찰하였다. 또한, SCLE를 처리한 A-549, AGS 및 MCF-7 암세포주에서 세포 주기의 G1기에서 세포 성장이 정지되었고,세포 사멸이 유의적으로 증가하는 것을 알 수 있었다. 그러나, SCLE 처리는 rho 단백질의 활성과 관련 없는 세포부착력의 소실과 세포 사멸이 유도되는 것을 관찰하였다. 이 연구의 결과를 바탕으로 토복령 추출물은 정상 세포보다는 암세포에 특이적으로 세포부착력의 소실과 세포 사멸을 유도하여, 이 추출물에 포함된 물질을 이용한 항암 연구에 응용될 수 있을 것으로 판단된다.

• 생명과학회지
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• 제30권12호
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• pp.1054-1062
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• 2020

벼메뚜기는 메뚜기과에 속하는 불완전 변태 곤충으로, 국내에서 오랫동안 식용으로 이용되어 왔다. 현재까지 벼메뚜기의 항산화 및 항염증 효과 등의 기능들이 연구되었으나, 탄수화물 소화 효소나 식후 혈당 수치에 미치는 영향에 관한 연구는 부족한 실정이다. 이에 본 연구는 벼메뚜기 에탄올 추출물(OEE)와 물 추출물(OWE)이 탄수화물 소화 효소를 저해하고, streptozotocin (STZ)으로 유도된 당뇨병 마우스에서 식후 고혈당을 강하시키는 효과에 대해 조사하였다. 그 결과 OEE에 의한 α-glucosidase와 α-amylase 저해 효과가 OWE 보다 더 효과적이었다. 당뇨병 마우스에 전분(2 g/kg)을 투여한 후의 혈당 증가는 15, 30, 60, 120분에 각각 406.50, 431.50, 473.50 and 460.00 mg/dl로 나타났고, 전분(2 g/kg)과 OEE 추출물(300 mg/kg)을 투여한 후 혈당 증가는 15, 30, 60, 120분에 각각 327.00, 361.00, 358.67 and 291.00 mg/dl로 나타나, OEE 추출물 투여군이 대조군에 비해 식후 혈당 강하가 효과적으로 나타남을 알 수 있었다. 이러한 결과는 벼메뚜기 추출물이 α-glucosidase와 α-amylas를 저해함으로써 식후 고혈당을 완화시키고, 특히 벼메뚜기 에탄올 추출물(OEE)이 벼메뚜기 물 추출물(OWE) 보다 식후 고혈당을 완화시키는데 더욱 효과가 있는 것으로 나타났다. 따라서 OEE가 탄수화물 소화 효소 저해 효과로 식후 고혈당을 완화시키는 유용한 천연 기능성 식품이 될 것으로 사료된다.

• 대한본초학회지
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• 제33권4호
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• pp.101-108
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• 2018

Objectives : This study aimed to effects antioxidant activity of citrus peel extract (CPE) and effect on its glucose metabolism in L6 rat skeletal muscle cells. Methods : Antioxidative activities were evaluated by using 10 kinds of natural materials, and total polyphenol and flavonoid contents were examined. The L6 muscle cells toxicity of CPE was examined by MTT assay. Expression of glucose-related genes in L6 muscle cells by CPE treatment was analyzed by real-time PCR and western blotting. Results : The $IC_{50}$ values of DPPH and ABTS free radical scavenging activity of CPE were ($15.47{\pm}0.26{\mu}g/m{\ell}$ and $12.07{\pm}1.23{\mu}g/m{\ell}$, respectively), effectively clearing DPPH and ABTS. CPE showed total polyphenol and flavonoid contents ($20.30{\pm}0.38$ and $64.20{\pm}0.52$, respectively). The selected CPE were used in experiments using an effective concentration that is not toxic in L6 muscle cells. We investigated insulin receptor substrate-1 (IRS-1), phosphatidylinositol 3-kinase regulatory (PI3KR), Akt, and glucose transporter 4 (GLUT4). mRNA analysis by realtime PCR showed no significant difference, but CPE-treated cells showed a tendency to increase in concentration-dependent manner. However, analysis of protein expression of Akt and GLUT4 by western blotting showed that CPE treatment significantly increased concentration dependent (p<0.001). Conclusions : As a result, citrus peel extract with high antioxidant activity regulates glucose metabolism in L6 muscle cells. Therefore, CPE can be a potential treatment for the treatment of diabetes.

• 한국수산과학회지
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• 제56권1호
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• pp.7-20
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• 2023

Boil-dried concentrate (BDC) and steam-dried concentrate (SDC) were prepared from olive flounder Paralichthys olivaceus roe using the cook-dried process, and their food functionality and in vitro bioactivity were examined. The buffer capacity of BDC and SDC was found to be stronger in the alkaline region than in the acidic region, and the buffer capacity of SDC was superior to that of BDC. The water holding capacities of these concentrates were 7.6 and 7.4 g/g protein, respectively, both of which were significantly lower than that of freeze-dried concentrate (FDC). The solubility of BDC (13.4%) and SDC (12.7%), foaming capacity of BDC (107.7%) and SDC (110.6%), and oil-in-water emulsifying activity index of BDC (7.7 m²/g) and SDC (9.7 m²/g) were all significantly lower than the corresponding values for FDC (P<0.05). The lower food functionality of BDC and SDC compared with FDC can be attributed to the high-temperature denaturation of proteins during the cook-dried process. The 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid radical scavenging activities (IC₅₀) of SDC (2.5 mg protein/mL) was 60.4 ㎍/mL, and the angiotensin I converting enzyme inhibitory activity was 80.9%. Olive flounder roe concentrates have good antioxidant and antihypertensive activities, and can be used as materials or ingredients in the processing of seafood and other foods to enhance protein contents and food functionality.

• 대한한방부인과학회지
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• 제22권1호
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• pp.1-14
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• 2009

Purpose: Oxidative stress was thought to play a critical role in neurodegenerative disease. Many in vivo and in vitro reports explained the possible pathway of human aging. But in therapeutic aspects, there was no clear answers to prevent aging associated with neural diseases. In this study, we investigated the antioxidant and neuroprotective effects of the Insamyangyung-tang (IYT). Methods: To estimate the antioxidant effects, we carried out 1.1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging assay, 2,2'-azinobis-(3- ethylbenzothiazoline-6- sulfonic acid (ABTS) radical cation decolorization assay, and measurement of total polyphenolic content. To evaluate neuroprotective effect of IYT in vitro. We performed thiazolyl blue tetrazolium bromide (MTT) assay, reactive oxygen species (ROS) creation in SH-SY5Y. Tyrosine hydroxylase (TH) immunocytochemistry, nitric oxide (NO) assay, and TNF-${\alpha}$ assay in primary rat mesencephalic dopaminergic neurons. Results: The $IC_{50}$ values were $571.6{\mu}g/m{\ell}$ and $202.3{\mu}g/m{\ell}$ in DPPH and ABTS assay respectively. Total polyphenolic content was 1.05%. In SH-SY5Y culture, IYT significantly increased the decreased cell viability by 6-OHDA at the concentrations of $10{\mu}g/m{\ell}$ in pre-treatment group, $10-100{\mu}g/m{\ell}$ in post-treatment group, and $100{\mu}g/m{\ell}$ in co-treatment group. The production of ROS induced by 6-OHDA was significantly inhibited in IYT treated group. In mesencephalic dopaminergic cell culture, the IYT group reduced the dopaminergic cell loss against 6-OHDA toxicity and the production of No and TNF-${\alpha}$ at the concentration of $0.2{\mu}g/m{\ell}$. Conclusion: These results showed that IYT has antioxidant and neuroprotectctive effects in the dopaminergic cells through decreasing the production of ROS, NO and TNF-${\alpha}$ which can cause many neurodegenerative changes in brain cell.

• 한국해양바이오학회지
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• 제15권1호
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• pp.1-11
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• 2023

In the present study, the antioxidant and anti-inflammatory activities of ethanolic extracts from Lathyrus japonicus at concentrations of 50, 100, and 200 ㎍/mL were investigated in LPS-stimulated, RAW264.7 cells. Antioxidant properties were determined using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging assays and ferric reducing antioxidant power assay. In addition, the production of reactive oxygen species (ROS) was measured using the 2',7'-dichlorofluorescein diacetate (DCFH-DA) probe by flow cytometry. To examine the anti-inflammatory activity of the extracts of L. japonicus, their effects on the levels of nitric oxide (NO); production of cytokines such as interleukin (IL)-1β, IL-10, and tumor necrosis factor-α (TNF-α); and the activities of enzymes such as inducible NOS (iNOS) and cyclooxygenase-2 (COX-2) were assessed. The IC₅₀ values of the DPPH and ABTS radical scavenging assays were 476.09 ± 1.50 and 34.91 ± 0.37 ㎍/mL, respectively. In addition, L. japonicus extracts not only inhibited ROS production, but also the production of NO, IL-1β, and IL-10, and the activity of iNOS in a dose-dependent manner. In summary, the ethanolic extracts of L. japonicus could be used as a functional food additive and an anti-inflammatory agent owing to their antioxidant and anti-inflammatory activities

• Biomolecules & Therapeutics
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• 제32권4호
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• pp.481-491
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• 2024

Paxlovid is the first approved oral treatment for coronavirus disease 2019 and includes nirmatrelvir, a protease inhibitor targeting the main protease (M^pro) of SARS-CoV-2, as one of the key components. While some specific mutations emerged in M^pro were revealed to significantly reduce viral susceptibility to nirmatrelvir in vitro, there is no report regarding resistance to nirmatrelvir in patients and animal models for SARS-CoV-2 infection yet. We recently developed xenograft tumors derived from Calu-3 cells in immunodeficient mice and demonstrated extended replication of SARS-CoV-2 in the tumors. In this study, we investigated the effect of nirmatrelvir administration on SARS-CoV-2 replication. Treatment with nirmatrelvir after virus infection significantly reduced the replication of the parental SARS-CoV-2 and SARS-CoV-2 Omicron at 5 days post-infection (dpi). However, the virus titers were completely recovered at the time points of 15 and 30 dpi. The virus genomes in the tumors at 30 dpi were analyzed to investigate whether nirmatrelvir-resistant mutant viruses had emerged during the extended replication of SARS-CoV-2. Various mutations in several genes including ORF1ab, ORF3a, ORF7a, ORF7b, ORF8, and N occurred in the SARS-CoV-2 genome; however, no mutations were induced in the M^pro sequence by a single round of nirmatrelvir treatment, and none were observed even after two rounds of treatment. The parental SARS-CoV-2 and its sublineage isolates showed similar IC₅₀ values of nirmatrelvir in Vero E6 cells. Therefore, it is probable that inducing viral resistance to nirmatrelvir in vivo is challenging differently from in vitro passage.

• 한국산업보건학회지
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• 제34권2호
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• pp.166-178
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• 2024

Objectives: The 12 occupational exposure limits(OELs) for chromium and its compounds in Korea were set by applying the American Conference of Governmental Industrial Hygienists (ACGIH) Threshold Limit Values (TLVs). However, this is significantly different from the TLVs after the existing TLVs were integrated and withdrawn in 2018, so it is necessary to review the revision. Methods: Various documents related to chromium OELs were reviewed, including the ACGIH TLV Documentations for chromium and its compounds. A field survey was conducted targeting workplaces handling chromium and its compounds. Based on this, a revised OELs were proposed and a socio-economic evaluation was conducted. Results: The OELs for chromium compounds in Korea was first enacted in 2002, and in 2007, the OELs for chromium (hexavalent) compounds (insoluble) was lowered from 0.05 mg/m³ to 0.01 mg/m³. In 2008, the OELs for strontium chromate was newly established as 0.0005 mg/m³, and in 2018, the OELs for calcium chromate was newly established as 0.001 mg/m³. Total chromium and hexavalent chromium were measured for each of 6 samples at 2 welding sites, 4 plating sites, and 2 spray coating sites. When omparing the average of the results measured by ICP, a total chromium analysis method, and the analysis results by IC, a hexavalent chromium analysis method, only workplace 4 was the same, and total chromium was evaluated more, and total chromium was evaluated at 0.0004 to 0.0027 mg/m³. And hexavalent chromium was evaluated as non-detection ~ 0.0014 mg/m³. Amendment ①: The exposure standard for hexavalent chromium is not divided into water soluble, insoluble, chromium ore processing, and other hexavalent chromium compounds, and is integrated into 0.01 mg/m³, which is the level of chromium (hexavalent) compound (insoluble)., OELs for chromium (metal) and chromium (trivalent) compounds are integrated into chromium (trivalent) compounds, and the exposure level is maintained. Amendment ②: As in the amendment ①, the OELs are integrated, but the level is lowered to 0.005 mg/m³, which is the OELs of OSHA, and there is a grace period of 4 years. Amendment ③: As in the amendment ①, the OELs are integrated, but the level is lowered to 0.0002 mg/m³, which is the exposure standard of ACGIH, and there is a grace period of 5 years. Conclusions: Amendment ①: The change in the OELs is insignificant, so the cost required is small, and the benefit/cost ratio is greater than 1, so there is no problem in applying the amendment. Amendment ②: In all scenarios except chromium 6(insoluble), the benefit/cost ratio is greater than 1, so it is thought that there will be no major problem in applying the amendment. Amendment ③: Since the benefit/cost ratio is less than 1 in all scenarios, it is thought that the total social benefit that can be obtained when applying the amendment is not large.

• 한국식품영양과학회지
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• 제40권12호
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• pp.1642-1647
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• 2011

본 연구는 왕쥐똥나무잎(Ligustrum ovalifolium H.) 추출물의 항산화 활성을 탐색하고자 물과 에탄올로 각각 추출하였다. 왕쥐똥나무잎에 포함된 총 폴리페놀과 총 플라보노이드 함량을 측정한 결과, 물 추출물($105.5{\pm}1.31$ mg GAE/g extract)과 에탄올 추출물($102.1{\pm}1.82$ mg GAE/g extract)은 비슷한 총 폴리페놀 함량을 함유하고 있었으며, 총 플라보노이드 함량 또한 물 추출물($84{\pm}1.72$ mg CE/g extract)과 에탄올 추출물($82.8{\pm}1.65$ mg CE/g extract)이 비슷한 양을 함유하고 있었다. ESR을 이용한 라디칼 소거능을 측정한 결과, hydroxyl 라디칼을 제외한 DPPH, alkyl 라디칼 소거능은 물 추출물 및 에탄올 추출물 모두 높은 활성을 나타내었다. 특히 alkyl 라디칼 소거능은 비타민 C와 같은 활성을 나타내었다. ABTS를 이용한 라디칼 소거활성, FRAP을 이용한 총항산화능 측정 및 환원력을 통한 항산화 활성을 측정한 결과에서도 왕쥐똥나무잎 추출물이 뛰어난 항산화효과를 가지고 있음이 확인되었다. 특히, FTC 및 TBA법을 이용한 지질과산화 억제 효능을 살펴본 결과, 왕쥐똥나무잎 물추출물과 에탄올 추출물 모두 지질과산화 억제효과가 높았으며 ${\alpha}$-토코페롤보다 활성이 훨씬 우수하였다. 한편, 세포독성을 살펴보기 위하여 정상 간세포(human liver, Chang cells)를 이용하여 MTT assay를 통한 세포의 생존율을 살펴본 결과 물 추출물 및 에탄올 추출물 모두 0.5 mg/mL의 농도까지 전혀 독성을 나타내지 않았다. 따라서 이 연구를 바탕으로 앞으로 왕쥐똥나무잎을 이용한 천연 항산화제로서의 가능성에 대한 추가적인 연구가 필요할 것으로 판단된다.