• Asian Pacific Journal of Cancer Prevention
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• 제17권12호
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• pp.5139-5145
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• 2016

There has been an increment in the number of studies focused on marine bioactive materials. Many peptides and other biomaterials with anticancer potential have been extracted from various marine animals. Artemia extracts have found uses in sun-light protection cosmetics and anti-aging products. However, contents of biochemical compounds in Artemia spp. and molecular mechanisms of have not been clearly studied in leukemic cells in vitro. In this work, we isolated and purified proteins of Artemia Urmiana. Six clear fractions (A-F) observed on DEAE-cellulose chromatography were assayed for effects on cell growth, differentiation and apoptosis using the human leukemic HL-60 cell line. Cell proliferation analysis by MTT and BrdU assays indicated that did not affect cells, growth. Cells treated with crude extract and fractions A, B and C, but not E and F (up to $100{\mu}g/mL$), exhibited increase of cell growth in a dose dependent manner. Stimulatory effects of fraction D were observed at concentrations of $10{\mu}g/mL$ and above. In nitro blue tetrazolium (NBT) reduction assays, treatment with $100{\mu}g/mL$ of fraction E or F for 96 hr increased the fraction of differentiated cells up to $14.8{\pm}3.56%$ and $16.5{\pm}2.08%$ respectively. Combination of those fractions with retinoic acid had significant synergistic effects on the differentiation of cells ($56.8{\pm}3.7%$ and $67.4{\pm}4.2%$, $p{\leq}0.01$). Annexin-V FITC staining for apoptosis and flow cytometric assays indicated induction of apoptosis by fractions E and F up to 23.8 and 31.8% of cells.

• Journal of Oral Medicine and Pain
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• 제26권4호
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• pp.319-329
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• 2001

Endothelin-1 (ET-1) is a recently discovered potent vasoconstrictive peptide. It was first identified in vascular endothelial cells. ET-1 is a 21-amino acid peptide and elicits systemic effects such as stimulation of the production of atrial natriuretic peptide and release of aldosterone and corticosterone. In this study, to examine the role of ET-1 in the bone metabolism, effect of ET-1 on the proliferation and activity of osteoblastic cells was studied using HOS cells as osteoblast model. ET-1 dose-dependently increased the cell proliferation as determined by cell counting and MTT reduction assay after 48hr treatment. Alkaline phosphatase activity was inhibited by ET-1 and showed significant inhibition by 50 and 100 nM ET-1. ET-1 increased NBT reduction by HOS cells dose-dependently showing that ET-1 may increase the superoxide production by osteoblasts. Nitrite concentration in the media of HOS cell culture without cytokine stimulation was negligible and unaffected by ET-1 after 48hr treatment. Finally, after collection and concentration of conditioned media, gelatinase activity produced by HOS cells was determined by zymography. HOS cells can produce and secrete the gelatinase (gelatinase A type as determined by molecular weight of about 65,000) into culture media, however, ET-1 had no effect on the gelatinase activity. These findings suggest that ET-1 may have diverse effects on the proliferation and differentiation of osteoblasts, therefore, it may play an important role in bone metabolism.

• BMB Reports
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• 제39권6호
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• pp.722-729
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• 2006

Recently, we have reported that 3-hydrogenkwadaphnin (3-HK), a diterpene ester isolated from Dendrostellera lessertii (Thymealeaceae), is very effective against leukemia cell lines without any detectable effects on normal cells (Moosavi et al., 2005b). In this study, we report that 3-HK induces $G_1$ cell-cycle arrest, differentiation and apoptosis in APL NB4 cell line. Indeed, the drug between 24 to 96 h induced 7-65% growth inhibition of NB4 cells. Cell viability was also decreased by 2-55% between 24 to 96 h treatments with the drug, respectively. These effects of the drug were also dose-dependent. According to flow cytomtry results, 3-HK (15 nM) induced a significant G1-arrest up to 24 h which was consequently followed with appearance of sub-$G_1$ peak at 72 to 96 h. Hoechst 33258 staining and DNA fragmentation assays confirmed the occurrence of apoptosis among the treated cells. On the other hand, NBT reducing assay, Wright-Giemsa staining, phagocytic activity and expression of cell surface markers (CD11b and CD14) confirmed that the inhibition of proliferation is associated with differentiation especially toward macrophage-like morphology. Interestingly, 3-HK at 5 and 10 nM enhanced the effects of all-trans retinoic acid (ATRA) in NB4 cells. Based on these results, 3-HK might become an ideal candidate for treatment of APL patients pending full exploration of its biological functions.

• Journal of Animal Science and Technology
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• 제56권1호
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• pp.3.1-3.7
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• 2014

We examined the effects of Prunella vulgaris Labiatae (P. vulgaris L.) on specific and non-specific immune responses of Nile tilapia, Oreochromis niloticus. The optimal concentration without toxicity of P. vulgaris was determined to $30-40{\mu}g/ml$ in vitro and $120{\mu}g$/100 g of fish in vivo. P. vulgaris significantly elicited an antibody titer compared to FCA or ${\beta}$-glucan. ${\beta}$-glucan plus P. vulgaris group synergistically enhanced antibody production. No significant difference in antibody production was observed between P. vulgaris and P. vulgaris plus ${\beta}$-glucan group. A respiratory burst activity of head kidney (HK) leucocytes of tilapia administered with 300 or $500{\mu}g$ P. vulgaris was significantly (p < 0.05) enhanced compared with the PBS-injected control group and FCA-treated group. Maximum increase in the NBT reduction value was observed in $500{\mu}g$ P. vulgaris group but no significant difference was found between 300 and $500{\mu}g$ P. vulgaris group. The level of serum lysozyme activity was significantly (p < 0.05) higher in the 300 and $500{\mu}g$ P. vulgaris than $100{\mu}g$ P. vulgaris and FCA group. The phagocytic activities of HK leucocytes from tilapia administered with 300 and $500{\mu}g$ P. vulgaris were significantly (p < 0.05) higher than $100{\mu}g$ P. vulgaris and the control group. P. vulgaris was revealed with a good immunoadjuvant evoking the specific and non-specific immune responses of tilapia.

• 한국약용작물학회지
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• 제6권4호
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• pp.311-322
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• 1998

60종(種)의 자생 약용식물과 식용작물을 대상으로 항산화활성의 생리활성을 조사함으로써 약용식물과 식용작물의 유용성 측면의 확인뿐만 아니라 새로운 생리활성물질 탐색의 가능성을 검토하기 위해서 실시한 실험 결과는 다음과 같다. 자생 약용식물 및 식용작물에 대한 50% EtOH 추출물을 이용하여 TBA법. DPPH법에 의하여 1차 활성검정 결과 처리방법에 따라 활성값에 차이가 있으나 TBA법에서는 검정콩의 추출액이 87.3%, DPPH법에서는 까마중 추출액이 80.6%로서 가장 높은 활성을 보였고 두가지 방법에서 동시에 활성이 높은 것은 검정콩을 비롯하여 10종 이었다. SOD활성정도도 검정식물에 따라 큰 차이가 있으나 검정콩이 53.5%로서 가장 높은 SOD활성을 나타내었으며 SOD종류는 Cu/ZnSOD, FeSOD을 포함하는 것으로 나타났다. Cu/ZnSOD만을 함유하고 있는 잔대는 SOD활성 정도가 10.37%로서 실험 대상식물 중 가장 낮은 활성을 보였다.

• 한국어병학회지
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• 제21권3호
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• pp.219-228
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• 2008

The effects of dietary yeast β-glucan administration on growth, nonspecific immune responses, serum lysozyme, skin mucous lysozyme, NBT (nitroblue tetrazolium) reduction by phagocytes, and disease resistance against Edwardsiella tarda in Japanese eel, Anguilla japonica were evaluated. Fish were fed the diets supplemented with 0%, 0.1% and 0.5% of yeast β-glucan to a commercial diet for 6 weeks. The body weight gain from the fish fed on the 0.5% supplemented diet for 6 weeks was significantly higher than the control. Both serum and skin mucous lysozyme were significantly higher in the all experimental groups except 2 weeks of 0.5% group. The bactericidal activity of serum was slightly increased at 6 weeks. Also, The intracellular superoxide anion production of kidney phagocytes was significantly higher in the all experimental groups. The diet supplemented with 0.1% were also found to raise the relative percent survival (RPS) of Japanese eel after an artificial challenge with 1×107 cells of Edwardsiella tarda per fish. The results suggested the potential of yeast β-glucan to activate some innate immune responses and to improve the growth in Japanese eel.

• The Korean Journal of Physiology and Pharmacology
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• 제2권4호
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• pp.479-491
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• 1998

In order to know the pathogenesis of adult respiratory distress syndrome (ARDS) in association with the oxidative stress by neutrophils, the role of platelet activating factor (1-0-alkyl-2-acetyl-snglycero-3-phosphocholine, PAF) was investigated during acute lung injury induced by interleukin- $1{\alpha}$ (IL-1) in rats. An insufflation of IL-1 into the rat's trachea increased the acetyltransferase activity in the lung and the increase of PAF content was followed. As evidences of acute lung injury by neutrophilic respiratory burst, lung leak index, myeloperoxidase activity, numbers of neutrophils in the bronchoalveolar lavage fluid, neutrophilic adhesions to endothelial cells and NBT positive neutrophils were increased after IL-1 treatment. In addition, a direct instillation of PAF into the trachea caused acute lung leak and the experimental results showed a similar pattern in comparison with IL-1 induced acute lung injury. For the confirmation of oxidative stress during acute lung leak by IL-1 and PAF, a histochemical electron microscopy was performed. In IL-1 and PAF treated lungs of rats, the deposits of cerrous perhydroxide were found. To elucidate the role of PAF, an intravenous injection of PAF receptor antagonist, WEB 2086 was given immediately after IL-1 or PAF treatment. WEB 2086 decreased the production of hydrogen peroxide and the acute lung leak. In ultrastructural study, WEB 2086 mitigated the pathological changes induced by IL-1 or PAF. The nuclear factor kappa B (NFkB) was activated by PAF and this activation was inhibited by WEB 2086 almost completely. Based on these experimental results, it is suggested that the PAF produced in response to IL-1 through the remodeling pathway has the major role for acute lung injury by neutrophilic respiratory burst. In an additional experiment, we can also come to conclude that the activation of the NFkB by PAF is thought to be the fundamental mechanism to initiate the oxidative stress by neutrophils causing release of proinflammatory cytokines and activation of phospholipase $A_2$.

• Korean Journal of Acupuncture
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• 제28권2호
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• pp.23-36
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• 2011

목적 : 이 연구는 관절 및 심혈관계 질환 치료에 사용되는 송절(松節)(Pinus densiflora Gnarl)을 약침용 시료로 조제하여 본 약물의 항산화 효능을 규명하고자 하였으며 이를 다양한 시스템에서 검토하였다. 방법 : $FeCl_2$-ascorbic acid system에서 흰쥐 간조직의 지질과산화 반응을 관찰하였고, Fenton reaction system에서 자유기에 의한 plasmid DNA 분절을 유도하였다. 또한 deoxyribose assay를 통해 hydroxyl radical 소거능을 관찰하였고, NBT reduction assay로 superoxide radical 소거능을 검토하였다. 또한 human low-density lipoprotein(LDL)의 산화를 유도하기 위해 $CuSO_4$와 AAPH를 사용하였으며 relative electrophoretic mobility (REM) assay로 LDL 산화 억제 효능을 대조 항산화물질과 비교 검토하였다. 결과 : 송절 약침액은 자유기에 의한 간조직의 지질과산화(p < 0.01)및 DNA 분절을 현저하게 억제하였으며, hydroxyl radical, superoxide radical (p < 0.01), nitric oxide 및 peroxynitrite를 강하게 소거하였다. 또한 $CuSO_4$ ($IC_{50}=9.2{\pm}0.2\;{\mu}g/ml$)와 AAPH ($IC_{50}=34.8{\pm}5.1\;{\mu}g/ml$)에 의해 유도된 human LDL의 산화를 억제하였고, REM assay에서도 산화 억제 효능을 재확인할 수 있었다. 결론 : 송절 약침액은 활성산소종 및 활성질소종를 소거하였고, 지질과산화를 억제하였으며, 특히 human LDL의 산화적 손상을 방어하였다. 이에 본 약물은 자유기에 의한 심혈관의 산화적 손상을 효과적으로 보호할 것으로 판단된다.

• 동의생리병리학회지
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• 제20권5호
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• pp.1315-1320
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• 2006

The root and rhizomes of Nardostachys chinensis belonging to the family Valerianaceae has been used for medicinal therapy in Korean traditional medicine. The parts have been especially used to elicit stomachic and sedative effects. Our previous studies reported that the water extract of N. chinensis has induced granulocytic differentiation inhuman promyelocytic leukemia (HL-60) cells. The Mitogen-activated protein kinases (MAPKs) are serine/threonine kinases involved in the regulation of various cellular responses, such as cell proliferation, differentiation and apoptosis. In this study, we investigated the signaling pathways on the HL-60 cell differentiation induced by N. chinensis. Activation of extracellular signal-regulated kinase (ERK) increased time-dependently in differentiation of HL-60 cells induced by N. chinersis. Activation of p38 increased slightly at 24 h after N. chinensis treatment, but activation of c-jun N-terminal kinase (JNK) was unaffected. Inhibitor of ERK (PD98059) significantly reduced NBT reduction activity induced by N. chinensis in HL-60 cells. In contrast, p38 inhibitor (SB203580) did not inhibit the cell differentiation. These results indicated that activaiton of ERK may De involved in HL-60 cell differentiation induced by N. chinensis.

• Preventive Nutrition and Food Science
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• 제9권3호
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• pp.283-288
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• 2004

Superoxide dismutase (SOD) from tomato (Lycopersicon esculentum Mill.) fruit was purified by ammonium sulphate precipitation, Sephadex G-100 and DEAE-cellulose column chromatographies. A 22 fold purification and an overall yield of 44% were achieved. The purified enzyme was a homodimer with Mr 37.1 kDa and subunit Mr 18.2 kDa as judged by SDS-PAGE. SOD showed $K_{m}$ values of 25 ${\times}$ 10$^{-6}$ M and 1.7 ${\times}$ 10$^{-6}$ M for nitroblue tetrazolium (NBT) and riboflavin as substrates, respectively. The enzyme was thermostable upto 5$0^{\circ}C$ and exhibited pH optima of 7.8. The effect of metal ions and some other compounds on enzyme activity was studied. $Co^{2+}$ and $Mg^{2+}$ were found to enhance relative enzyme activities by 27 % and 73 %, respectively, while M $n^{2+}$ inhibited the SOD activity by 64%. However, $Ca^{2+}$ and C $u^{2+}$ had no effect on enzyme activity. Other compounds like $H_2O$$_2$ and Na $N_3$ inhibited enzymatic activities by 60% and 32%, respectively, while sodium dodecyl sulphate (SDS), chloroform plus ethanol and $\beta$-mercaptoethanol had no effect on the activity of SOD. of SOD.