• 한국식품영양과학회지
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• 제46권8호
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• pp.919-928
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• 2017

큰잎모자반에 함유되어 있는 다당류를 저분자화하기 위해 알긴산 분해 효소를 생산하는 Shewanella oneidensis PKA 1008의 조효소액을 첨가하여 저분자화하고 이의 면역증진능을 확인하였다. 먼저 저분자화됨을 확인하기 위하여 TLC를 실시한 결과 분해 24 h 이후부터 분해되기 시작하여 최종 60 h에서 dimers로 분해되었다. 그 후 비장세포에 큰잎모자반의 효소적 추출 분해물을 처리하여 in vitro에서 면역증진능을 확인한 결과 $IFN-{\gamma}$, IL-2, IL-6, IL-10의 경우 분해가 진행될수록 분비량이 많아졌다. 또한, 큰잎모자반의 효소적 추출 분해물을 2주 동안 마우스에 경구투여 한 결과, MTT assay 및 비장세포에서 분비되는 $IFN-{\gamma}$ 및 IL-2의 분비량의 경우 24 h 처리구에서 대조군보다 높은 수치로 농도 의존적으로 증가함을 나타내었다. IgG2a 분비량의 경우 24 h와 48 h 처리구에서 다소 증가하였으며, NK 세포의 경우 24 h 처리구에서 농도 의존적으로 활성이 증가하여 면역력을 증진시키는 결과를 보여주고 있다. 전혈을 이용하여 일반혈액검사의 경우 24 h 및 48 h 처리군에서 대조군보다 높은 수치를 나타내었다. 이상의 결과를 종합해볼 때 S. oneidensis PKA 1008 유래 조효소에 의한 큰잎모자반 효소적 추출 분해물이 면역 관련 세포증식률을 증가시키고 이에 따른 사이토카인의 분비량을 증가시키며 혈액 내 다양한 면역세포들의 수치를 증가시킴으로써 면역증진능에 탁월한 효과를 나타냄을 확인하였으나, 면역증진 효과에 기여하는 활성물질에 대해서는 밝혀지지 않아 그에 대한 추가적인 연구가 요구된다.

• 한국약용작물학회지
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• 제17권5호
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• pp.311-320
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• 2009

The low quality fresh ginseng was fermented by Phelinus linteus or Hericium erinaceum mycelium. This fermented ginseng was extracted by water at $100^{\circ}C$ or water with ultrasonification at $60^{\circ}C$. Total phenolic compounds was improved by ultrasonification extraction process, compare to conventional water extraction. All extracts enhanced the growth of human B and T cells, showing 2.68 times and 3.43 times higher, respectively, than the control. The secretion of TNF-$\alpha$ and IL-6 from human immune cells was enhanced as $3.53{\times}10^{-4}\;pg/cell$, $3.40{\times}10^{-4}\;pg/cell$ by adding H. erinaceum mycelium fermented ginseng. H. erinaceum mycelium fermented ginseng yielded higher nitric oxide production from macrophage than Lipopolysaccharides (LPS). The cytotoxicity on human normal kidney cell (HEK293) was as low as 20.5% in adding the maximum concentration of $1.0\;mg/m{\ell}$ of fermented ginseng. Generally, the extracts from ultrasonification extraction process showed 10% lower toxicity than that by conventional process. H. erinaceum mycelium fermented ginseng had the highest anticancer activity on human lung cancer and stomach cancer cells as 69.33% and 75.32%, respectively at $1.0\;mg/m{\ell}$. It can be concluded that, in general, H. erinaceum mycelium fermented ginseng has relatively better immune and anticancer activities than P. linteus fermented ginseng. Expecially, the extracts treated with ultrasonification had higher activities than that from conventional extraction process.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2003년도 생물공학의 동향(XIII)
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• pp.195-199
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• 2003

물 추출과 초음파를 병행한 모든 조건 중 $60^{\circ}C$에서 초음파 병행 추출물이 보다 높은 추출 수율을 보였고 그 중 복분자가 가장 높은 추출 수율을 보였다. 또한 총 5일간 인간 면역세포인 T-cell의 생육도와 Cytokine 분비량을 확인한 결과도 마찬가지로 물 추출물보다는 초음파를 병행한 추출물을 더 높은 면역 증진 효과를 보였다. 이는 초음파를 통해 발생되는 액체 상호간의 상승작용으로 인하여 생약재중에 함유되어 있는 많은 phenol성 물질 및 식물성분의 증가로 생리활성 물질의 증가를 가져오기 때문이다. 더욱이 $100^{\circ}C$가 아닌 $60^{\circ}C$에서 더 높은 면역 증진활성을 보인 이유는 액체 상태의 과도한 열로 인하여 내부 식물성분의 파괴 및 열 변성을 가져와 이런 결과를 가져온 것으로 생각된다. 이상의 결과로 기존의 $100^{\circ}C$ 물 추출물보다는 $60^{\circ}C$의 비교적 낮은 온도에서 초음파 추출을 통한 생약재의 추출물들이 더 높은 활성을 나타내므로 초음파 추출에 대한 다양한 실험 모델을 통해 더 많은 실험이나 기능성 식품 및 소재 개발에 이용되고자 한다.

• 대한치과교정학회지
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• 제28권6호
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• pp.915-926
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• 1998

Cytoskeleton은 세포핵과 세포외 기질을 연결하고 있어서 기질에 가해지는 물리적 힘에 의해 cytoskeletal change가 유도되고 이에 의해 세포의 개조활성이 영향을 받는다고 생각되어 왔다. 본 연구는 골모세포 활성에 대한cytoskeletal change의 역할을 규명하기 위한 것으로서, 신생 백서로부터 조골세포양 세포를 분리, 배양하고 네가지 농도의 cytochalasin B(CB) 또는 colchicine(COL)을 3시간 처리하였다. 다시 배양액을 교환하고 24시간 동안 배양하여 prostaglandin $E_2\;(PGE_2)$, interleukin-6(IL-6), tumor necrosis factor-$\alpha$(TNF-$\alpha$) 및 matrix metalloproteinase-1 (MMP-1) 생산을 측정하고 통계적으로 비교하였으며 cytoskeletal protein actin 변화를 관찰하기위하여 면역형광염색하고 형광현미경으로 관찰하여 다음과 같은 결과를 얻었다: 1. CB 처리군에서 $PGE_2$ 생산이 증가되는 경향을 보였고 COL 처리군에서는 약물농도에 비례하여 증가하였다. 2. IL-6 생산은 CB농도 1.0 ${\mu}g/ml$일때를 제외하고 증가되었다. 3. TNF-$\alpha$도 CB 농도가 1.0 ${\mu}g/ml$ 일때를 제외하고 증가하였다. 4. MMP-1 생산은 CB 처리군에서 감소하는 경향을 보이고 COL 처리군에서는 변화되지 않았다. 5. CB처리군에서는 cytoskeletal actin stress fibers가 사라지고 세포모양이 둥글어지는 경향을 보였다. 이상의 결과로 미루어 보아 cytoskeletal rearrangement는 골모세포유사세포의 활성, 특히 $PGE_2$, IL-6, 및 TNF-$\alpha$같은 paracrine/autocrine factor의 생산과 관련있는 것으로 보인다.

• Mycobiology
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• 제40권3호
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• pp.181-188
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• 2012

This study was initiated in order to investigate the anticancer and immunomodulating activities of crude polysaccharides extracted in methanol, neutral saline, and hot water (hereinafter referred to as Fr. MeOH, Fr. NaCl, and Fr. HW, respectively) from the fruiting bodies of Panellus serotinus. Content of ${\beta}$-glucan and protein in Fr. MeOH, Fr. NaCl, and Fr. HW extracts of P. serotinus ranged from 22.92~28.52 g/100 g and 3.24~3.68 g/100 g, respectively. In vitro cytotoxicity tests, none of the various fractions of crude polysaccharides were cytotoxic against sarcoma 180, HT-29, NIH3T3, and RAW 264.7 cell lines at the tested concentration. Intraperitoneal injection with crude polysaccharides resulted in a life prolongation effect of 23.53~44.71% in mice previously inoculated with sarcoma 180. Treatment with Fr. HW resulted in an increase in the numbers of spleen cells by 1.3 fold at the concentration of $50{\mu}g/mL$ compared with control. Treatment with Fr. NaCl resulted in improvement of the immuno-potentiating activity of B lymphocytes by increasing the alkaline phosphatase activity by 1.4 fold, compared with control, at the concentration of $200{\mu}g/mL$. Among the three fractions, maximum nitric oxide ($13.48{\mu}M$) was recorded at $500{\mu}g/mL$ in Fr. HW. Production of tumor necrosis factor alpha, interleukin-$1{\beta}$, and interleukin-6 was significantly higher, compared to the positive control, concanavalin A, at the tested concentration. Therefore, treatment with crude polysaccharides extracted from the fruiting body of P. serotinus could result in improvement of antitumor activity.

• 한국식품저장유통학회지
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• 제13권6호
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• pp.769-773
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• 2006

녹차종자를 기능성 식품소재로서 이용하기 위하여 녹차 종자메탄올추출물의 생리활성을 녹차 메탄올추출물의 생리활성과 비교하였다. 녹차 메탄올추출물기 수소공여능은 $100{\mu}g/mL$ 농도 이상에서 50% 이상의 활성을 나타내었으나, 녹차종자 메탄올추출물은 $1000{\mu}g/mL$ 농도에서 21.86%의 활성을 나타내었다. 또한 녹차종자 및 녹차 메탄올추출물이 $1000{\mu}g/mL$ 농도로 처리된 흰쥐의 간 균질물에서 MDA의 생성량이 대조구의 86 Mol/g에 비하여 각각 60 및 50 Mol/g로 낮게 나타내어 이들 추출물은 항산화효과가 있음을 알 수 있었다. 녹차종자 및 녹차 메탄올추출물은 $1000{\mu}g/mL$ 농도에서 A549 및 SW480 세포에 대하여 각각 20%, 50% 이상의 성장 억제율을 나타내었다. 또한 녹차종자 메탄을 추출물이 $500{\mu}g/mL$의 농도 처리된 암세포는 대조구에 비하여 상대적인 세포 수의 감소되었으며, 심한 형태학적인 변화를 보여주었다. 대식세포 RAW264.7에 녹차 및 녹차종자 메탄을 추출물이 1, 10, 100및 $1000{\mu}g/mL$의 농도로 처리 하였을 때, 이들 추출물은 $100 {\mu}g/mL$ 농도까지 농도 의존적으로 NO(nitric oxide)의 생성을 유도하였으며, $100{\mu}g/mL$의 농도에서 그 농도는 각각 0.77와 2.04 uM로서 녹차종자 메탄을 추출물이 녹차 메탄을 추출물보다 그 효과가 크게 나타났다.

• IMMUNE NETWORK
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• 제10권4호
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• pp.135-143
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• 2010

Background: Cordyceps militarys water extract (CME) has been reported to exert antitumor and immunomodulatory activities in vivo and in vitro. However, the therapeutic mechanism has not yet been elucidated. In this study, we examined the effects of CME on the antigen presenting function of antigen presenting cells (APCs). Methods: Dendritic cells (DCs) were cultured in the presence of CME, and then allowed to phagocytose microspheres containing ovalbumin (OVA). After washing and fixing the efficacy of OVA, peptide presentation by DCs were evaluated using CD8 and CD4 T cells. Also, we confirmed the protein levels of proinflammatory cytokines through western blot analysis. Results: CME enhanced both MHC class I and class II-restricted presentation of OVA in DCs. In addition, the expression of both MHC class I and II molecules was enhanced, but there was no changes in the phagocytic activity of exogenous OVA. Furthermore, CME induced the protein levels of iNOS, COX-2, proinflammatory cytokines, and nuclear p65 in a concentration-dependent manner, as determined by western blot. Conclusion: These results provide an understanding of the mechanism of the immuno-enhancing activity of CME on the induction of MHC-restricted antigen presentation in relation to their actions on APCs.

• 약학회지
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• 제54권6호
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• pp.494-499
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• 2010

We have been focussing on discovery of natural compounds that have immunoregulatory activities for many years. In the present study, we investigated if chlorogenic acid (CRA), a polyphenolic compound, has an immunoadjuvant activity. Prior to examining the immunoadjuvant activity, effect of CRA on proliferation of T- or B-lymphocyte was determined. Results showed that CRA enhanced the proliferation of those lymphocytes in dose-dependant manner (P<0.05), and the proliferation enhancement by CRA was appeared to be more effective to B-cells than to T-cells. Based on these observations, it was tested with bovine serum albumin (BSA) and Candida albicans cell wall (CACW) as antigenic sources if CRA has an immunoadjuvant activity. In experiments, BSA alone or a mixture of BSA plus CRA was injected intraperitoneally to mice (BALB/c strain). For a negative control, mice were given only diluent (DPBS) by the same route. In other experiment, CACW was tested by the same way as did with BSA. Three weeks after the first immunization these animals were boosted. Antisera collected from the mice one week after the booster were analyzed by ELISA. Results displayed that the induction of anti-BSA antibody was increased in mice that received the mixture of BSA and CRA as compared to anti-BSA induction in BSA only-given mice groups (P<0.05). In case of CACW, a similar observation as did with BSA was made, resulting in that there was app. 40% increased production of the anti-CACW antiserum from the combination (CACW plus CRA)-received mice as compared to antiserum induction from CACW alone-given animals. Taken all together, these data indicate that CRA has an ability of enhancing antibody production regardless of nature of antigenic sources. Presumably, activation of B-cell proliferation by CRA may plays an important role in the immunoadjuvant activity of the polyphenolic compound.

• International Journal of Industrial Entomology and Biomaterials
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• 제7권2호
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• pp.165-173
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• 2003

Cytokines represent an essential part of the innate immune response in mammals. Recently, several studies have reported the presence of cytokine-like activities and molecules in the invertebrates such as echinoderms, tunicates, mollusks and insects. In our serial study, we investigated presence of cytokines in the silkworm, Bombyx mori, infected with several immune inducers. Western blotting analysis using rabbit anti-human cytokines showed the presence of IL-6-like molecule in the hemolymph collected at 8 and 24 hrs after infection with peptidoglycan and oligodeoxynucleotide, and the molecular weight of the proteins was ∼45 kDa. We attempted to isolate the molecule by gel permeation HPLC, anion exchange chromatography, ultra centrifugation, and immuno-dot-blot assay, but until now the effort was not much successful yet. It, however, does not appear that the IL-6-like molecule in the silkworm larvae is a mere experimental artifact happened by Western blotting analysis. Instead, further experiment on this subject probably will provide us more fruitful result as detected in other invertebrates including insects.

• 대한한의학회지
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• 제31권1호
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• pp.93-111
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• 2010

Objectives: The present study was performed to evaluate the potential effects of Bupleuri radix (SH) on regenerative activities in the peripheral sciatic nerve after crushing injury in rats. Methods: Axonal regeneration after crush injury in rats was analyzed by immunofluorescence staining using anti-NF-200 antibody and retrograde tracing of DiI-axons. Changes in protein levels in the sciatic nerve axons and DRG tissue were analyzed by Western blot analysis and immunofluorescence staining. Effects of SH extract treatment on neurite outgrowth was examined by immunofluorescence staining for cultured DRG neurons. Results: Major findings on the effects of SH extract treatment on axonal regeneration are summarized as follows. 1. SH-mediated enhancement in axonal regeneration was identified by immuno- fluorescence straining of NF-200 protein and retrograde tracing of DiI-labeled axons. 2. Axonal GAP-43 protein levels were upregulated by SH not only in the injured axons but also in the DRG sensory neurons corresponding to sciatic sensory axons. 3. Phospho-Erk1/2 protein levels were increased in both injured axonal area and DRG sensory neurons by SH. Phospho-Erk1/2 was also found in non-neuronal cells in the injured axons. 4. SH elevated levels of Cdc2 protein produced in Schwann cells in the distal portions of injured sciatic nerves. 5. The neurite outgrowth of DRG sensory neurons in culture was augmented by SH, and these changes were positively associated with GAP-43 production levels in the DRG neurons. Conclusions: These data suggest that SH extract improves the regenerative responses of injured peripheral neurons, and thus may be useful for understanding molecular basis for the development of therapeutic strategies.