• Archives of Pharmacal Research
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• 제29권10호
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• pp.859-865
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• 2006

With an approach to study the anti-tumor effects and mechanism of selenium compound, we investigated the anti-tumor activity and mechanism of $Na_5SeV_5O_{18}{\cdot}3H_2O$ (NaSeVO) in K562 cells. The results showed that $0.625{\sim}20\;mg/L$ NaSeVO could significantly inhibit the proliferation of K562 cells in vitro in a time- and concentration-dependent manner as determined by microculture tetrazolium (MTT) assay, the IC50 values were 14.41 (4.45-46.60) and 3.45 (2.29-5.22) mg/L after 48 hand 72 h treatment with NaSeVO respectively. In vivo experiments demonstrated that i.p. administration of 5, 10 mg/kg NaSeVO exhibited an significant inhibitory effect on the growth of transplantation tumor sarcoma 180 (S180) and hepatoma 22 (H22) in mice, with inhibition rate 26.8% and 58.4% on S180 and 31.3% and 47.4% on H22, respectively. Cell cycle studies indicated that the proportion of G0/G1 phase was increased at 2.5 mg/L while decreased at 10 mg/L after treatment for 24, 48 h. Whereas S phase was decreased at 2.5-5 mg/L and markedly increased at 10 mg/L after treatment for 48 h. After treatment for 24 h, 10 mg/L NaSeVO also markedly increased S and G2/M phases. Take together, the result clearly showed that NaSeVO markedly increased S and G2/M phases at 10 mg/L. The study of immunocytochemistry showed that the expression bcl-2 is significantly inhibited by 10 mg/L NaSeVO, and bax increased. Morphology observation also revealed typical apoptotic features. NaSeVO also significantly caused the accumulation of $Ca^{2+}$ and $Mg^{2+}$, reactive oxygen species (ROS) and the reduction of pH value and mitochondrial membrane potential in K562 cells as compared with control by confocal laser scanning microscope. These results suggest that NaSeVO has anti-tumor effects and its mechanism is attributed partially to apoptosis induced by the elevation of intracellular $Ca^{2+}$, $Mg^{2+}$ and ROS concentration, and a reduction of pH value and mitochondria membrane potential (MMP).

• 한국식품영양과학회지
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• 제40권3호
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• pp.366-371
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• 2011

본 연구는 남해 통흑마늘, 의성 통흑마늘, 창녕 통흑마늘 및 본 연구실에서 개발한 제조방법으로 만든 흑마늘의 이화학적 특성 및 항산화 특성을 비교하였다. 각각 흑마늘의 당도는 42.7, 42.7, 56.7, $54.7^{\circ}Brix$를 나타내었다. 환원당 함량은 각각 15.0, 16.0, 23.5, 25.9%로 본 연구실에서 제조한 흑마늘의 환원당 함량이 가장 높게 나타났다. 시판 및 본 연구실에서 제조한 발효숙성 흑마늘의 pH는 3.9~4.6으로 생마늘을 흑마늘로 만드는 과정에서 산도가 높았다. Thiol의 함량은 각각 448.5, 350.0, 318.2, 478.7 mg/g으로 본 연구실에서 개발한 방법으로 제조한 흑마늘의 thiol 함량이 가장 높게 나타났다(p<0.05). 총 phenol 함량은 각각 0.64, 0.60, 0.68, 0.72 mg/mL로 본 연구실에서 제조한 흑마늘이 가장 높았다. FRAP은 각각 1.7, 1.4, 1.7, 1.9 mg/mL로 본 연구실에서 개발한 제조방법으로 만든 발효숙성 흑마늘의 이 가장 높게 나타났다. ABTS 라디칼 소거능은 73.8, 66.2, 95.8, 96.0%로 본 연구실에서 개발한 방법으로 제조한 흑마늘 항산화능이 가장 높게 나타났다. DPPH 및 hydroxyl 라디칼 소거능의 $IC_{50}$(DPPH 라디칼을 50% 소거시키는데 필요한 농도)값은 각각 13.1, 12.4, 12.4, 9.1 mg/mL 및 5.2, 3.7, 4.0, 3.2 mg/mL로 본 연구실에서 개발한 방법으로 제조한 흑마늘의 $IC_{50}$이 가장 낮아, 항산화능이 가장 높게 나타났다. 위 결과로부터 본 연구실에서 제조한 흑마늘은 당도, 환원당, thiol 함량 및 항산화능이 시판되는 흑마늘보다 높아 건강식품소재로 우수함을 나타내었다.

• 한국식품영양과학회지
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• 제43권12호
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• pp.1843-1851
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• 2014

다시마에는 항산화성과 무기질이 풍부하고 알긴산 등 증점 다당류가 풍부한 해조류이다. 다시마 추출물 함량을 달리한 재래식 보리된장을 제조하여 2개월간 발효시키면서 항산화능 및 이화학적 실험을 실시하였다. 제조된 된장의 pH는 5.80~6.86, 산도는 0.57~1.87%, 수분 함량은 65.30~40.90% 범위로 저장기간 동안 pH는 감소하였고 산도는 증가하였다. 점도는 보리된장 대조군이 4,913 cps이었고 다시마 추출물의 함량이 증가할수록 4,793~9,333.3 cps까지 비례하여 증가하였으며 저장기간에 따라 비례하였다(P<0.05). 다시마를 첨가한 보리된장의 색도는 다시마 추출물의 첨가량에 따라 각각 L 값과 b 값은 증가하였으나 a 값은 다시마 추출물의 함량에 따라 감소하는 경향을 나타내었다. 총 페놀 함량은 대조군이 12.72 mg TAE/g, 처리군이 13.40~16.37 mg TAE/g이었으며(P<0.05), flavonoid 함량은 대조군이 $0.98{\mu}gRE/g$, 처리군이 $1.25{\sim}1.56{\mu}gRE/g$으로 다시마 추출물의 첨가량과 비례하여 항산화능에 영향을 주었다(P<0.05). DPPH 라디칼 소거능은 BHA를 (+)대조군으로 하여 $IC_{50}$ 함량을 각각 구하였으며 보리된장 대조군 23.23 mg/mL, 다시마 추출물 처리군의 경우 24.83 mg/mL, 10.28 mg/mL 수준으로 다시마 추출물이 된장의 항산화능을 증진시킨 것으로 나타났다. 다시마 추출물을 첨가한 보리된장의 총균수는 초기에 7.20~7.57 log CFU/g으로 나타났고 초기 젖산균수는 4.20~4.71 log CFU/g 범위로 나타나 다시마 처리군의 총균수는 대조군보다 적고 젖산균 수는 더 많은 것으로 나타났다. 관능검사에서 감칠맛은 대조군에 비해서 20% 다시마 처리군이 6.1점으로 유의적인 차이를 보였고(P<0.05), 질감 또한 대조군(5.7점)보다 20% 다시마 추출물을 첨가한 군이 6.4점으로 유의적으로 높았으며(P<0.05), 전체적 기호도는 대조군이 5.6점이었고 20% 첨가 처리군에서 6.5점으로 유의차가 크게 나타났다(P<0.01). 이상의 결과에서 보리된장에 다시마 추출물을 첨가하면 항산화능도 뛰어나므로 보리된장의 제품화 가능성도 더욱 크다고 사료된다.

• Asian Pacific Journal of Cancer Prevention
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• 제14권11호
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• pp.6363-6367
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• 2013

Atractylis lancea (Thunb.) DC. (AL), an important medicinal herb in Asia, has been shown to have anti-tumor effects on cancer cells, but the involved mechanisms are poorly understood. This study focused on potential effects and molecular mechanisms of AL on the proliferation of the Hep-G2 liver cancer cell line in vitro. Cell viability was assessed by MTT test in Hep-G2 cells incubated with an ethanol extract of AL. Then, the effects of AL on apoptosis and cell cycle progression were determined by flow cytometry. Telomeric repeat amplification protocol (TRAP) assays was performed to investigate telomerase activity. The mRNA and protein expression of human telomerase reverse transcriptase (hTERT) and c-myc were determined by real-time RT-PCR and Western blotting. Our results show that AL effectively inhibits proliferation in Hep-G2 cells in a concentrationand time-dependent manner. When Hep-G2 cells were treated with AL after 48h,the $IC_{50}$ was about 72.1 ${\mu}g/mL$. Apoptosis was induced by AL via arresting the cells in the G1 phase. Furthermore, AL effectively reduced telomerase activity through inhibition of mRNA and protein expression of hTERT and c-myc. Hence, these data demonstrate that AL exerts anti-proliferative effects in Hep-G2 cells via down-regulation of the c-myc/hTERT/telomerase pathway.

• Archives of Pharmacal Research
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• 제27권4호
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• pp.390-395
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• 2004

The methanol extract obtained from the aerial parts of Aceriphyllum rossii (Saxifragaceae) was fractionated into ethyl acetate (EtOAc), n-BuOH and $H_2O$ layers through solvent fractionation. Repeated silica gel column chromatography of EtOAc and n-BuOH layers afforded six flavonol glycosides. They were identified as kaempferol 3-O-$\beta$-D-glucopyranoside (astragalin, 1), quercetin 3-O-$\beta$-D-glucopyranoside (isoquercitrin, 2), kaempferol 3-O-$\alpha$-L-rhamnopyranosyl $(1{\to}6)-\beta$-D-glucopyranoside (3), quercetin 3-O$\alpha$-L-rharnnopyranosyl $(1{\to}6)-\beta$-D-qlucopyrano-side (rutin, 4), kaempferol 3-O-[$\alpha$-L-rharnnopyranosyl $(1{\to}4)-\alpha$-L-rhamnopyranosyl $(1{\to}6)-\beta$-D-glucopyranoside] (5) and quercetin 3-O-[$\alpha$-L-rhamnopyranosyl $(1{\to}4)\alpha$-L-rhamnopyranosyl $(1{\to}6)\beta$-D-glucopyranoside] (6) on the basis of several spectral data. The antioxidant activity of the six compounds was investigated using two free radicals such as the ABTS free radical and superoxide anion radical. Compound 1 exhibited the highest antioxidant activity in the ABTS $\{2,2-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid)\}$ radical scavenging method. 100 mg/L of compound 1 was equivalent to $72.1\pm1.4\;mg/L$ of vitamin C, and those of compounds 3 and 5 were equivalent to $62.7\pm0.5\;mg/L$ and $54.3\pm1.3\;mg/L$ of vitamin C, respectively. And in the superoxide anion radical scavenging method, compound 5 exhibited the highest activity with an $IC_{50}$ value of $17.6{\pm}0.3{\mu}M$. In addition, some physical and spectral data of the flavonoids were confirmed.

• Asian Pacific Journal of Cancer Prevention
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• 제13권11호
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• pp.5455-5461
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• 2012

Luteolin is a plant flavonoid which exhibits anti-oxidative, anti-inflammatory and anti-tumor effects. However, the antiproliferative potential of luteolin is not fully understood. In this study, we investigated the effect of luteolin on cell cycling and apoptosis in human esophageal squamous carcinoma cell line Eca109 cells. MTT assays showed that luteolin had obvious cytotoxicity on Eca109 with an $IC_{50}$ of $70.7{\pm}1.72{\mu}M$ at 24h. Luteolin arrested cell cycle progression in the G0/G1 phase and prevented entry into S phase in a dose- and time-dependent manner. as assessed by FCM. Luteolin induced apoptosis of Eca109 cells was demonstrated by AO/EB staining assay and annexin V-FITC/PI staining. Moreover, luteolin downregulated the expression of cyclin D1, survivin and c-myc, and it also upregulated the expression of p53, in line with the fact that luteolin was able to inhibit Eca109 cell proliferation.

• Natural Product Sciences
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• 제23권2호
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• pp.75-83
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• 2017

The optimization and microwave assisted extraction of stem bark of Terminalia arjuna, quantitative estimation of the marker compounds arjunic acid and arjunolic acid using HPTLC and the evaluation of free radical scavenging activity has been performed in this study. The central composite design was used for optimization and the values of parameters for optimized batch of microwave assisted extraction were 1000 W (Power), 3 minutes (Time) and 1/120 (Solid/solvent ratio). The solvent system to carry out the HPTLC was toluene: acetic acid: ethyl acetate (5: 5: 0.5) and quantitative estimation was done using standard equations obtained from the marker compounds. The in-vitro free radical scavenging activity was performed spectrophotometrically using ascorbic acid as standard. The value of estimated percentage yield of arjunic acid and arjunolic acid was 1.42% and 1.52% which upon experimentation was obtained as 1.38% and 1.51% respectively. The DPPH assay of the different batches of microwave assisted extraction and marker compounds taken suggested that the marker compounds arjunic acid and the arjunolic acid were responsible for the free radical scavenging activity as the batch having the maximum percentage yield of the marker compounds showed best free radical scavenging effect as compared to standard ascorbic acid. The $IC_{50}$ value of the optimized batch was found to be 24.72 while that of the standard ascorbic acid was 29.83. Hence, the yield of arjunic acid and arjunolic acid has direct correlation with the free radical scavenging activity of stem bark extract of Terminalia arjuna and have potential to serve as active lead compounds for free radical scavenging activity.

• Asian Pacific Journal of Cancer Prevention
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• 제16권6호
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• pp.2473-2481
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• 2015

Colon cancer is one of the leading causes of cancer-associated death worldwide. The prognosis for advanced colorectal cancers remains dismal, mainly due to the propensity for metastatic progression. Accordingly, there is a need for effective anti-metastasis therapeutic agents. Since a great body of research has indicated anticancer effects for curcumin, we investigated the effects of dendrosomal curcumin (DNC) on cellular migration and adhesion of human SW480 cells and possible molecular mechanisms involved. Different methods were applied in this study including MTT, Scratch and adhesion assays as well as real-time PCR and transwell chamber assays. Based on the results obtained, DNC inhibits metastasis by decreasing Hef 1, Zeb 1 and Claudin 1 mRNA levels and can reduce SW480 cell proliferation with $IC_{50}$values of 15.9, 11.6 and $7.64{\mu}M$ at 24, 48 and 72h post-treatment. Thus it might be considered as a safe formulation for therapeutic purpose in colorectal cancer cases.

• Natural Product Sciences
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• 제25권4호
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• pp.326-333
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• 2019

The purpose of our study was to evaluate anti-AD potential of Cirsium maackii flowers. MeOH extract, CH₂Cl₂, EtOAc, and n-BuOH fraction of this flower notably inhibited BACE1 (IC₅₀ = 76.47 ± 1.66, 22.98 ± 1.45, 8.65 ± 0.63, and 72.47 ± 3.04 ㎍/mL, respectively). β-amyrenone (49.70 mg) (1), lupeol acetate (1.43 g) (2), lupeol (1.22 g) (3), lupenone (23.70 mg) (4), β-sitosterol (1.01 g) (6), and β-sitosterol glucoside (13.00 mg) (7) from CH₂Cl₂, apigenin (100.20 mg) (8), luteolin (19.00 mg) (9), apigenin 7-O-glucuronide methyl ester (21.30 mg) (14), and tracheloside (53.70 mg) (5) from EtOAc, apigenin 5-O-glucoside (11.00 mg) (10), luteolin 5-O-glucoside (11.00 mg) (11) and apigenin 7-O-glucuronide (91.00 mg) (12) from n-BuOH, and luteolin 7-O-glucuronide (22.00 mg) (13) from H₂O fraction were isolated. HPLC showed high levels of 8, 9 and 12 in MeOH extract (33.07 ± 0.07, 31. 44 ± 0.17 and 16.89 ± 0.33 mg/g, respectively), EtOAc (161.01 ± 1.78, 96.93 ± 0.34 and 73.38 ± 0.06 mg/g, respectively), and n-BuOH fraction (32.18 ± 0.33, 44.31 ± 0.32 and 105.94 ± 0.36 mg/g, respectively). Since, 3 and 9 are well-known BACE1 inhibitors, the anti-AD activity of C. maackii flower might be attributable to their presence.

• Asian Pacific Journal of Cancer Prevention
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• 제14권2호
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• pp.631-637
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• 2013

Luteolin is a naturally occurring flavonoid present in many plants with diverse applications in pharmacology. Despite several studies elucidating its significant anti-cancer activity against various cancer cells, the mechanism of action in skin cancer is not well addressed. Hence, we investigated the effects of luteolin in HaCaT (human immortalized keratinocytes) and A375 (human melanoma) cells. The radical scavenging abilities of luteolin were determined spectrophotometrically, prior to a cytotoxic study (XTT assay). Inhibitory effects were assessed by colony formation assay. Further, the capability of luteolin to induce cell cycle arrest and apoptosis were demonstrated by flow cytometry and cellular DNA fragmentation ELISA, respectively. The results revealed that luteolin possesses considerable cytotoxicity against both HaCaT and A375 cells with $IC_{50}$ values of 37.1 ${\mu}M$ and 115.1 ${\mu}M$, respectively. Luteolin also inhibited colony formation and induced apoptosis in a dose and time-dependent manner by disturbing cellular integrity as evident from morphological evaluation by Wright-Giemsa staining. Accumulation of cells in G2/M (0.83-8.14%) phase for HaCaT cells and G0/G1 (60.4-72.6%) phase for A375 cells after 24 h treatment indicated cell cycle arresting potential of this flavonoid. These data suggest that luteolin inhibits cell proliferation and promotes cell cycle arrest and apoptosis in skin cancer cells with possible involvement of programmed cell death, providing a substantial basis for it to be developed into a potent chemopreventive template for skin cancer.