• Preventive Nutrition and Food Science
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• 제20권1호
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• pp.22-28
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• 2015

Phenolic rich ethyl acetate fraction (EAF) from lotus leaves was prepared and its bioactive components, antioxidant and cytoprotective effects were investigated. EAF showed high total phenolic content and flavonoid content and contained rutin ($11,331.3{\pm}4.5mg/100g\;EAF$), catechin ($10,853.8{\pm}5.8mg/100g\;EAF$), sinapic acid ($1,961.3{\pm}5.6mg/100g\;EAF$), chlorogenic acid ($631.9{\pm}2.3mg/100g\;EAF$), syringic acid ($512.3{\pm}2.5mg/100g\;EAF$), and quercetin ($415.0{\pm}2.1mg/100g\;EAF$). EAF exerted the $IC_{50}$ of $4.46{\mu}g/mL$ and $5.35{\mu}g/mL$ toward DPPH and ABTS cation radicals, respectively, and showed strong reducing power, which was better than that of ascorbic acid, a positive control. Additionally, EAF protected hydroxyl radical-induced DNA damage indicated by the conversion of supercoiled pBR322 plasmid DNA to the open circular form and inhibited lipid peroxidation of polyunsaturated fatty acid in a linoleic acid emulsion. In cultured hepatocytes, EAF exerted a cytoprotective effect against oxidative stress by inhibiting intracellular reactive oxygen species formation and membrane lipid peroxidation. In addition, depletion of glutathione under oxidative stress was remarkably restored by treatment with EAF. The results suggest that EAF have great potential to be used against oxidative stress-induced health conditions.

• Asian Pacific Journal of Cancer Prevention
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• 제16권17호
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• pp.7943-7957
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• 2015

Background and Aims: Colorectal cancer is one of the leading causes of death in the world. The aim of this study was to investigate the growth-suppression potentiality of a crude saponin extract (CSENS) prepared from medicinal herb, Nigella sativa, on human colon cancer cells, HCT116. Materials and Methods: HCT116 cells were subjected to increasing doses of CSENS for 24, 48 and 72 h, and then harvested and assayed for cell viability by WST-1. Flow cytometry analyses, cell death detection ELISA, fluorescent stains (Hoechst 33342 and acridine orange/ethidium bromide), DNA laddering and comet assays were carried out to confirm the apoptogenic effects of CSENS. Luciferase reporter gene assays, quantitative reverse transcription-polymerase chain reaction and Western blot analyses were performed to assess the impact of CAERS and CFEZO on the expression levels of key regulatory proteins in HCT116 cells. Results: The results demonstrated that CSENS inhibited proliferation and induced apoptosis. Apoptosis was confirmed by flow cytometry analyses, while CSENS-treated cells exhibited morphological hallmarks of apoptosis including cell shrinkage, irregularity in cellular shape, cellular detachment and chromatin condensation. Biochemical signs of apoptosis, such as DNA degradation, were observed by comet assay and gel electrophoresis. The pro-apoptotic effect of CSENS was caspase-3-independent and associated with increase of the Bax/Bcl-2 ratio. CSENS treatment down-regulated transcriptional and DNA-binding activities of NF-${\kappa}B$ and AP-1 proteins, associated with down-regulation of their target oncogenes, c-Myc, cyclin D1 and survivin. On the other hand, CSENS up-regulated transcriptional and DNA-binding activities of Nrf2 and expression of cytoprotective genes. In addition, CSENS modulated the expression levels of ERK1/2 MAPK, p53 and p21. Conclusions: These findings suggest that CSENS may be a valuable agent for treatment of colon cancer.

• Asian Pacific Journal of Cancer Prevention
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• 제17권4호
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• pp.1947-1959
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• 2016

Background: Colorectal cancer (CRC) is a major cause of morbidity and mortality, being the second most common type of cancer worldwide in both men and women. It accounts yearly for approximately 9% of all new cases of cancers. Furthermore, the current chemotherapeutic regimens seem unsatisfactory, so that exploration of novel therapeutic modalities is needed. The present study was undertaken to investigate the inhibitory effects of a crude alkaloid extract (CAERS) of a medicinal herb, Rhazya stricta, on proliferation of CRC HCT116 cells and to elucidate mechanisms of action. To achieve these aims, we utilized MTT, comet, DNA laddering and gene reporter assays, along with Western blot and RT-PCR analyses. Results: We found that CAERS inhibited cell proliferation and induced apoptotic cell death in HCT116 cells. Hallmarks of morphological and biochemical signs of apoptosis were clearly evident. CAERS down-regulated DNA-binding and transcriptional activities of NF-${\kappa}B$ and AP-1 proteins, while up-regulating expression of the Nrf-2 protein. It also down-regulated expression levels of the ERK MAPK, Bcl-2, cyclin D1, CDK-4, survivin and VEGF and up-regulated levels of Bax, caspase-3/7 and -9, p53, p21, Nrf-2. Markedly, it promoted mRNA expression levels of cytoprotective genes including the hemeoxygenase-1, NAD(P)H quinine oxidoreductase 1 and UDP-glucuronyltransferase. Conclusions: These findings indicate that CAERS exerts antiproliferative action on CRC cells through induction of apoptotic mechanisms, and suggest CAERS could be a promising agent for studying and developing novel chemotherapeutic agents aimed at novel molecular targets for the treatment of CRC.

• 대한화장품학회지
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• 제43권2호
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• pp.87-92
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• 2017

본 연구는 머위 추출물의 화장품소재로서의 가능성을 확인하기 위하여 자체의 독성과 카드뮴으로부터 유도된 세포손상에 미치는 영향에 대하여 평가하였다. 카드뮴으로부터 손상을 유도한 각질형성세포에 머위 추출물을 처리하여 세포사멸 인자인 Bcl-2와 procaspase-3의 단백질 발현을 측정하였다. 그 결과 머위 추출물 $200{\mu}g/mL$를 제외한 모든 농도에서 98% 이상의 높은 생존율을 나타내었으며 세포사멸인자인 Bcl-2와 procaspase-3 단백질 발현이 증가한 것으로 보아 머위 추출물이 카드뮴 독성 시 일어나는 세포자멸사에 대한 보호기전을 나타낸 것으로 평가되었다. 또한 카드뮴으로 12 h 동안 PARP cleavage를 유도한 각질형성세포에 머위 추출물을 전처리한 결과, 카드뮴을 매개로 하는 PARP cleavage를 억제하는 것으로 나타났다. 이러한 결과를 통해 머위 추출물이 피부세포 보호 효능을 나타내는 천연소재로서의 활용가능성을 제안한다.

• Journal of Microbiology and Biotechnology
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• 제26권2호
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• pp.277-286
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• 2016

Makgeolli lees (ML) has several physiological effects such as antioxidant, antidiabetic, and anticancer properties, but its biological functions have not been determined definitively. Here, we tested whether ML has a cytoprotective effect on paraquat (PQ)-induced oxidative stress in the human lung carcinoma cell line A549. At 0.1 mg/ml ML, viability of PQ-exposed A549 cells was restored by 12.4%, 18.5%, and 48.6% after 24, 48, and 72 h, respectively. ML also reduced production of the intracellular reactive oxygen species (ROS) that were generated by PQ treatment. Further experiments revealed that ML treatment enhanced the expression and nuclear translocation of nuclear factor erythroid 2-related factor 2 (NRF2) as well as ARE-GFP reporter activity. ML treatment also effectively increased the expression of NRF2's target genes NAD(P)H dehydrogenase quinone 1 (NQO1) and heme oxygenase 1 (HO-1). Moreover, we found that expression of cytoprotective genes, including glutathione peroxidases (GPXs), superoxide dismutase (SOD1), catalase (CAT), peroxiredoxin 3 (PRDX3), and peroxiredoxin 4 (PRDX4), was greatly enhanced by treatment with ML during PQ exposure. Taken together, the data suggest that treatment of PQ-exposed A549 cells with ML ameliorates cytotoxicity through induction of NRF2 expression and its target genes HO-1, NQO1, and other antioxidant genes. Thus, ML may serve as a functional food applicable to ROS-mediated human diseases.

• 생명과학회지
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• 제27권3호
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• pp.310-317
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• 2017

본 연구진은 HaCaT-ARE-luciferase 세포를 이용하여 400 여개의 약용식물 에탄올 추출물 중 NRF2/ARE 유도효과가 있는 신규 추출물을 검색하였고 이를 통하여 종대황(Rheum undulatum)과 선복화(Inula japonica)의 주정 추출물이 HaCaT-ARE-luciferase 세포에서 ARE 활성을 강하게 유도하는 것을 관찰하였다. 종대황과 선복화 에탄올 추출물은 HaCaT 세포에서 생존(viability)을 증가시켰고 NRF2/ARE에 의존적인 phase II cytoprotective 효소인 heme oxygenase-1 (HO-1)와 NADPH:quinone oxidoreductase-1 (NQO1)의 전사 및 단백질 발현을 강하게 유도하였다. 또한 종대황과 선복화 추출물은 HaCaT 세포에서 TPA로 유도한 세포 내 활성 산소 및 이를 통하여 생성되는 스트레스 마커인 8-hydroxydeoxyguanosine (8-OH-dG)과 4-hydroxynonenal (4HNE)의 발생을 강하게 억제하였다. 본 연구는 종대황과 선복화의 에탄올 추출물이 인간 피부 세포주인 HaCaT 세포에서 NRF2/ARE에 의존적인 유전자 발현의 유도를 통하여 강력한 항산화 효과를 발휘한다는 것을 증명한다.

• Natural Product Sciences
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• 제13권1호
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• pp.33-39
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• 2007

Eleven polyhydroxyursane triterpenoids (PHUTs) were tested to determine their cytoprotective, immunosuppressive and anti-inflammatory effects. To compare the bioactivities of $19{\alpha}$-hydroxyursane-type triterpenoids {23-hydroxytormentic acid (6), its methyl ester (7), tormentic acid (8), niga-ichigoside $F_1$ (9),euscaphic acid (10) and kaji-ichigoside $F_1$ (11)} of the Rosaceae crude drugs (Rubi Fructus and Rosa rugosae Radix) with PHUTs possessing no $19{\alpha}-hydroxyl$ of Centella asiatica (Umbelliferae), the four PHUTs, asiaticoside (1), madecassoside (2), asiatic acid (3), and madecassic acid (4) were isolated from C. asiatica and 23-hydroxyursolic acid (5) from Cussonia bancoensis. Cytoprotective effects were assessed by measuring cell viabilities against cisplatin-induced cytotoxocity in $LLC-PK_1$, cells (proximal tubule, pig kidney) to determine whether these agents have protective effects against nephrotoxicity caused by cisplatin. The inhibitory effect of 11 PHUTS on nitric oxide (NO) and prostaglandin $E_2\;(PGE_2)$ were evaluated by measuring nitrite accumulation in lipopolysaccharide (LPS)-induced macrophage RAW 264.7 cells, and their anti-inflammatory effects were tested in 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced mouse ear edema model. Six MHUTs (compounds 1, 2, 4, 6, 10, and 11) exhibited higher cell viabilities during cisplatin-induced cytotoxicity testing even at a concentration of $200\;{\mu}g/ml$ than cisplatin only-treated group, suggesting that ese compounds have the potentcytoprotective efffcts. Compounds 1 and 3 of the C. asiatica and niga-ichigoside $F_1$ exhibited no inhibitory effect on NO and/or $PGE_2$ production whereas other PHUTs produced mild to significant NO and/or $PGE_2$ production.The four compounds (2, 5, 9, and 10) potently inhibited mouse ear edema induced by TPA whereas two compounds (1 and 3) had no activity in this test. These results suggest that many PHUTs are potentchemopreventives. Structure-activity relationship (SAR) was also discussed in each assay with regard to the significant role of OHs at the position of 2, 3, 6, 19, and 23 and to the glycoside linkage at the 28-carboxyl.

• 동의생리병리학회지
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• 제21권1호
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• pp.28-32
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• 2007

Cisplatin, an antitumor agent widely used in the treatment of cancers, has nephrotoxicity. This side effect is closely related to oxidative stress. In the present study, we studied to protective effect of ethanol extract of Bojungbangam-tang (EBJT) on cisplatin-induced nephrotoxicity. Bojungbangam-tang is a new herbal prescription composed of nine crude drugs. Pretreatment of EBJT prevented cisplatin-induced cytotoxicity and generation of ROS. Also, cellular GSH content and gluathione peroxidase activity were recovered by EBJT. EBJT also decreased cisplatin-induced expression of HO-1 via inhibition of ERK activation. Taken together, these results suggest that EBJT has a cytoprotective effect against cisplatin-induced nephrotoxicity through anti-oxidant activity.

• Food Science and Biotechnology
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• 제17권5호
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• pp.1086-1091
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• 2008

The antioxidant activities of the extracts of dangyuja (Citrus grandis Osbeck) leaves were evaluated. The highest phenolic content was obtained from the ethyl acetate fraction (EF) (202.1$\pm$0.8 mg GAE/g dried extract) and it exhibited the highest 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging activity. The cytoprotective effects of EF on oxidative damage induced by tert-butyl hydroperoxide (t-BHP) in a human hepatoma cell line, HepG2 cells, were investigated to understand the intracellular antioxidant mechanisms. Treatment of HepG2 cells with EF prior to oxidative stress was found to inhibit reactive oxygen species (ROS) generation, lipid peroxidation, and DNA damage in a dose-dependent manner. Gas chromatography-mass spectrometry (GC-MS) studies on EF resulted in tentative identification of 19 compounds representing 94.3% of the total content. Taken together, these results demonstrated that EF has excellent antioxidant activities and thus dangyuja leaves have great potential as a source for natural antioxidant which can be applied in food products.

• Biomolecules & Therapeutics
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• 제24권4호
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• pp.426-432
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• 2016

Age-related rotator cuff tendon degeneration is related to tenofibroblast apoptosis. Anthocyanins reduce oxidative stress-induced apoptotic cell death in tenofibroblasts. The current study investigated the presence of cell protective effects in cyanidin and delphinidin, the most common aglycon forms of anthocyanins. We determined whether these anthocyanidins have antiapoptotic and antinecrotic effects in tenofibroblasts exposed to $H_2O_2$, and evaluated their biomolecular mechanisms. Both cyanidin and delphinidin inhibited $H_2O_2$-induced apoptosis in a dose-dependent manner. However, at concentrations of $100{\mu}g/ml$ or greater, delphinidin showed cytotoxicity against tenofibroblasts and a decreased antinecrotic effect. Cyanidin and delphinidin both showed inhibitory effects on the $H_2O_2$-induced increase in intracellular ROS formation and the activation of ERK1/2 and JNK. In conclusion, both cyanidin and delphinidin have cytoprotective effects on cultured tenofibroblasts exposed to $H_2O_2$. These results suggest that cyanidin and delphinidin are both beneficial for the treatment of oxidative stress-mediated tenofibroblast cell death, but their working concentrations are different.