• Journal of Animal Science and Technology
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• 제63권5호
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• pp.1159-1168
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• 2021

Ovotransferrin (OTF), an egg protein known as transferrin family protein, possess strong antimicrobial and antioxidant activity. This is because OTF has two iron binding sites, so it has a strong metal chelating ability. The present study aimed to evaluate the improved immune-enhancing activities of OTF hydrolysates produced using bromelain, pancreatin, and papain. The effects of OTF hydrolysates on the production and secretion of pro-inflammatory mediators in RAW 264.7 macrophages were confirmed. The production of nitric oxide (NO) was evaluated using Griess reagent and the expression of inducible nitric oxide synthase (iNOS) were evaluated using quantitative real-time polymerase chain reaction (PCR). And the production of pro-inflammatory cytokines (tumor necrosis factor [TNF]-α and interleukin [IL]-6) and the phagocytic activity of macrophages were evaluated using an ELISA assay and neutral red uptake assay, respectively. All OTF hydrolysates enhanced NO production by increasing iNOS mRNA expression. Treating RAW 264.7 macrophages with OTF hydrolysates increased the production of pro-inflammatory cytokines and the phagocytic activity. The production of NO and pro-inflammatory cytokines induced by OTF hydrolysates was inhibited by the addition of specific mitogen-activated protein kinase (MAPK) inhibitors. In conclusion, results indicated that all OTF hydrolysates activated RAW 264.7 macrophages by activating MAPK signaling pathway.

• 대한의생명과학회지
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• 제26권4호
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• pp.296-301
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• 2020

The purpose of this study was performed to evaluate antioxidant activity of the Artemisia asiatica Nakai and Moringa oleifera Lam mixture extract. Mixture extracts were manufactured by concentration and compared with a single extract (only the Artemisia asiatica Nakai mixture and only the Moringa oleifera Lam mixture). The experiments conducted Total polyphenol measurements, Total flavonoid measurements, DPPH radical scavenging activty, ABTS radical scavenging activty and LDH assay. The LDH assay assessment shows that all extracts are cells compared to controls. The toxicity was weak. Finally, The antioxidant capacity was rated higher than mixture extract of a single extract. Also, the optimized mixture was determined AM5 (Artemisia asiatica Nakai mixture: Moringa oleifera Lam mixture = 3:1). For extracts of AM5, Total phenol and flavonoid contents were 271.769±18.087 mg/g and 45.384±5.026 mg/g. and DPPH and ABTS scavenging activity were 70.8±6.496% and 77.1±9.634%. Therefore, it is expected that the value of the extract will increase as it increases its antioxidant activity if it is manufactured according to the appropriate ratio.

• Journal of Applied Biological Chemistry
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• 제65권2호
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• pp.89-92
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• 2022

In this study, the effects of soil conditions on antioxidant activities of the aerial and underground parts of rosemary were assessed to determine the most effective soil conditions for cultivation. The antioxidant activity was the highest (51.58±2.93 ㎍/mL) when cultivated in the mixture of gardening soil and vermiculite using DPPH assay. The antioxidant activity of underground parts the highest (127.48±12.38 ㎍/mL) when cultivated in the mixture of soil, vermiculite, and perlite. ABTS assay showed that the antioxidant activity of aerial parts was 230.34±57.93 ㎍·mL^-1 when cultivated in the mixture of gardening soil and vermiculite and that of underground parts was 320.98±16.04 ㎍·mL^-1 when cultivated in the mixture of gardening soil, vermiculite, and perlite. The total phenolic content of aerial parts was the highest (155.25±2.96 mg GAE/g) when cultivated in the mixture of gardening soil. The total flavonoid content of aerial parts was the highest (67.32±5.27 mg QE/g) when cultivated in the mixture of gardening soil. Therefore, the mixture of gardening soil, vermiculite, and perlite is superior to gardening soil alone for cultivation of rosemary to increase its antioxidant activity as well as total phenolic and flavonoid content.

• BMB Reports
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• 제28권5호
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• pp.437-442
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• 1995

The potency of yeast-derived methionyl-free human growth hormone (rhGH), which was obtained by removal of the N-terminal Met from methionyl-hGH, was estimated by in vitro and in vivo assays. In radio-receptor assay where the binding affinity of growth hormone to the receptor was estimated, the recombinant hGH showed 2.9 international units (IU) per mg of specific activity. In contrast, pitUitary-derived human growth hormone had a slightly lower receptor binding activity (2.5 IU/mg) compared with recombinant growth hormone. For the in vivo assay, efficacy of rhGH was tested by use of hypophysectomized rats, in which pituitary organs were surgically removed, resulting in the termination of growth hormone secretion. The weight-increase in rats by the injection of rhGH was almost identical to the result obtained by the injection of the same amount of pituitary-derived (international standard) hGH. A comparision of the secondary structures of rhGH and rMet-hGH by circular dichroism spectrophotometer demonstrated that the removal of the methionyl residue from rMet-hGH did not exert any effect on the structure of the growth hormone. In conclusion, methionyl-free human growth hormone produced from yeast was highly potent in biological activity and maintained a legitimate three dimensional structure.

• 생약학회지
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• 제38권1호
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• pp.84-89
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• 2007

The present study describes the preliminary evaluation of the antioxidant activities and the neuroprotective effect of methanolic extracts from Psoralea corylifolia Linne (PCE). The antioxidant activities and neuroprotective effect of the PCE were evaluated by total phenolic contents (TPC), DPPH radical scavenging activity (RSA), reducing power (RP), MTT reduction assay, and LDH release assay. TPC, DPPH RSA, and RP of the extract at concentration of 100 ${\mu}g$ was 125.93 ${\mu}g$, 63.81%, 0.138, respectively, and those were concentration dependent. The treatment of PC12 and N18-RE-105 cells with various PCE concentrations under $H_2O_2$ resulted in the induction of protective effect in a dose-dependent manner, as determined by the results of an MTT reduction assay and LDH release assay. Therefore, these results suggest that PCE could be a new potential candidate as an antioxidant against neuronal diseases.

• 대한약침학회지
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• 제17권1호
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• pp.44-50
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• 2014

Objectives: This study was undertaken to isolate a fibrinolytic enzyme from the snake venom of Gloydius blomhoffii siniticus and to investigate its enzymatic characteristics and hemorrhagic activity as a potential pharmacopuncture agent. Methods: The fibrinolytic enzyme was isolated by using chromatography, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and fibrin plate assay. The characteristics of the enzyme were investigated using fibrin plate assay, protein hydrolysis analysis, and hemorrhage assay. Its amino acid composition was determined. Results: The fibrinolytic enzyme with the molecular weight of 32kDa (FE-32kDa) from Gloydius blomhoffii siniticus showed a fibrin hydrolysis zone at the concentration of 0.2 mg/mL in the fibrin plate assay. The fibrin hydrolysis activity of the enzyme was inhibited completely by ethylenediaminetetraacetic acid (EDTA), ethyleneglycoltetraacetic acid (EGTA), and 1, 10-phenanthroline, thiothreitol and cysteine, and partially by phenylmethanesulfonylfluoride (PMSF). Metal ions such as $Fe^{2+}$ and $Hg^{2+}$ inhibited the fibrin hydrolysis completely, but $Zn^{2+}$ enhanced it. FE-32kDa hydrolyzed ${\alpha}$-chain but did not hydrolyze ${\beta}$-chain and ${\gamma}$-chain of fibrinogen. High-molecular-weight polypeptides of gelatin were hydrolyzed partially into low-molecular-weight polypeptides, but the extent of hydrolysis was limited. FE-32kDa induced hemorrhage beneath back skin of mice at the dose of $2{\mu}g$. Conclusions: FE-32kDa is a ${\alpha}$-fibrin(ogen)olytic metalloprotease that requires $Zn^{2+}$ for fibrinolytic activity and causes hemorrhage, suggesting that the enzyme is not appropriate for use as a clinical pharmacopuncture.

• Journal of the Korean Wood Science and Technology
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• 제35권6호
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• pp.126-134
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• 2007

음나무의 새로운 용도 개발을 위하여 다양한 항산화 측정법을 이용하여 생리활성을 측정하였다. 즉, 음나무 수피 및 근피의 온수 추출물과 메탄올 추출물에 대한 세포 내 활성산소 농도를 측정하여 세포수준에서의 산화 스트레스 억제효과를 측정하였으며, DPPH법 및 NBT법을 이용하여 각 추출물의 화학적인 측면에서의 항산화력을 측정하여 이들 간의 차이를 비교하였다. NR assay와 MTT assay에 의해 측정한 세포독성 실험결과, 음나무 근피의 메탄올 추출물의 세포독성이 가장 강했으며, 50%의 세포 생존율을 나타내는 NR50과 MTT50은 각각 0.0018%와 0.0029%였다. 산화 스트레스 억제효과는 근피의 불용성 온수 추출물이 15분 처리에서 57.9%로 가장 우수하였다. DPPH법과 NBT법을 이용한 음나무 추출물에 대한 항산화 활성 측정결과, 각각 수피의 메탄올 추출물이 96% (0.1% 농도)이고, 수피의 불용성 온수 추출물이 95% (0.5% 농도)를 나타내어 우수한 항산화 활성을 나타내었다.

• 생약학회지
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• 제41권3호
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• pp.166-173
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• 2010

Oxidative stress to proteins, lipids, or DNA is higher in human autopsy tissue and in rodent models of a number of neurodegenerative conditions, including Alzheimer's and Parkinson's disease. On the basis of this information, we established a screening system using N18-RE-105 cells to identify therapeutic agents that can protect cells from glutamate toxicity. During the course of our screening program, we recently isolated the active compound 9-hydroxy-$\alpha$-tocopherone ($\alpha$-TP), which prevents glutamate-induced cell death, from Viola mandshurica. The chemical structure of $\alpha$-TP was identified using spectroscopic methods and by comparison with literature values. Antioxidant activity and protective effects of $\alpha$-TP were evaluated by DPPH radical-scavenging assay, morphological assay, MTT reduction assay, and lactate dehydrogenase (LDH) release assay. These results suggest that $\alpha$-TP could be a new potential chemotherapeutic agent against neuronal diseases.

• 생약학회지
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• 제35권1호통권136호
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• pp.6-15
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• 2004

This study was performed to evaluate the hemopoietic effects of 6 species of deer antlers from origins and parts in vitro. CD34 positive cells were isolated and confirmed the its population by FACS analysis. In a week liquid culture, there was any statistical significance between extracts of three parts of six species of deer antlers in the experiments as colony forming assay, proliferation assay, differentiation assay and observation of morphology. However, after 2 weeks- culture with extracts of three parts of six species of deer antlers, colonies were counted. six species of deer antlers, such as middle part of Korean nippon deer, upper part of Chinese nippon deer, upper part of Newzealand horse deer, middle part of Korea horse deer and middle part of Newzealand red deer, significantly increased the CFU-GM (colony forming unit garnulocyte-macrophage) of CD34 positive cells re1atεd to production of leucocytes such as eosinophil, basophil and neutrophil, while only middle part of Korea horse deer significantly increased the BFU-E (burst forming unit-erythroid) at 1 mg/ml seggesting progenting red blood cells (RBC). In the molecular study with CD34+ cells pretreated with cyclophosphamide, antagonist of hemopoietic activity, upper Part of Korean nippon deer and upper part of Chinese nippon deer effectively increased TPO involved in a late pathway of hematopoiesis just like in ELISA assay of IL-3, TPO and GM-CSF. Taken together, these results indicate exσacts of deer antler had some hemopoietic activity still proposing more clinical study and more basic mechanism research.

• 대한예방한의학회지
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• 제5권1호
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• pp.144-150
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• 2001

The cytotoxic activity of Cratalariae sessiliflorae on cultured NIH 3T3 fibroblasts and human oral epithelioid carcinoma cells (KB) were evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazoliumbromide (MTT) colorimetric method These fractions of methanol extract of Cratalariae sessiliflorae showed inhibitory effect in vitro in the milligram range against KB cell lines. In general, the antitumor activities of these fractions were does-dependent over the milligram range. The comparison of IC50 values of these fractions in tumor cell lines showed that their susceptibility to these fractions decrease in the following order: Fr. 4> Fr. 6> Fr. 10> Fr. 2> Fr. 11> Fr. 3> Fr. 8> Fr. 7> Fr. 9> Fr. 1> Fr. 5 by the MTT assay. These fractions were tested for their cytotoxic effects on NIH 3T3 fibroblasts using MTT assay. They exhibited potent cytotoxic activities in vitro in the milligram range against NIH 3T3 fibroblasts. In general, the cytotoxic activities of these fractions were does-dependent over the milligram range. The comparison of CD50 values of these fractions in NIH 313 fibroblasts shows that their susceptibility to these fractions in decrease the following order: Fr. 10> Fr. 9> Fr. 2 = Fr. 4> Fr. 8> Fr. 11> Fr. 1 = Fr. 7> Fr. 3> Fr. 5 = Fr. 6 by the MTT assay. These results suggests that fraction 5 has the most growth - inhibitory activity against KB cell lines.