• 대한본초학회지
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• 제24권3호
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• pp.161-167
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• 2009

Objectives : Maekmoondong-tang (MMDT), a Korean herbal medicine, has been used to treat severe dry cough in patients with bronchitis and pharyngitis. MMDT has been reported to have anti-inflammatory, anti-allergic, immunomodulatory, secretory-modulating, and metabolic regulatory actions. However, there are no evidence in regard to the effects of MMDT on carcinogenesis and metastasis. Here, we investigated the effects of 70% ethanol extract of MMDT on cell viability, apoptosis, and motility in human hepatocarcinoma HepG2 cells. Methods : Cell viability was measured using the CCK-8 assay, and the apoptosis induction was evaluated by caspase-3 activity. To detect apoptotic features, the cells treated with MMDT were stained with 4'-6-diamidino-2-phenylindole (DAPI). Cell motility was examined by Boyden chamber assay and Real-time Cell Index of Migration assay. Gelatin zymography also performed to measure matrix metalloproteinase (MMP)-2/9 activity. Results : We found that MMDT significantly inhibited cell proliferation and increased caspase-3 activity in a dose-dependent manner in HepG2 cells. Apoptotic features such as chromatin condensation and apoptotic bodies were observed in MMDT-treated cells by DAPI staining. MMDT also suppressed PMA-induced cell motility and activities of MMP-2/9. Conclusions : Our results exhibited that MMDT possess the anti-carcinogenetic and anti-metastatic activities via caspase-3 activation and down-regulation of cell motility and invasion in HepG2 cells. Therefore, these findings suggest that MMDT could be potentially applied to the prevention and treatment of cancer.

• Parasites, Hosts and Diseases
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• 제29권1호
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• pp.21-30
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• 1991

간흡충의 충체 부위별 항원성을 알아보고자 면역조직화차적 방법을 이용하여 간흡충의 소화기관, 생식기관, 배설기관, 흡반, 표피 등의 염색강도를 비교 관찰하였다. 간흡충 실험 감염 후 2~8주 된 토끼에서 담관 내 충체를 포함하는 간 조직을 얻어 중성 포르말린 용액에 고정하고 파라핀으로 포매한 다음 4 $\mu\textrm{m}$두께로 잘라 항원으로 이용하였다. 항-간흡충 항체(1차 항체)는 실험 감염 10주 된 고양이 혈청을 희석하여 사용하였고 peroBidaseconjugated goat anti-cat IgG를 2차 항체로 하여 간접 면역효소 염색을 시행하였다. 가장 적합한 1차 항체의 희석 농도는 1 : 1,000~1 : 2,000, 2차 항체의 희석 농도는 1 : 1,000이었다. 충체의 장 상피세포, 장 내용물 및 배설낭은 1차 항체 희석 농도 1 : 1,000~1 : 2,000에서 강한 양성 반응을 보인 반면, 자궁 및 일부 자궁 내 충란, 난관선, 웅성 생식기관 등은 1차 항체 회석 농도 1 : 1,000에서 미약한 양성 반응을 나타내었다. 한편, 흡반, 표피, 표피하 세포, 충체 실질 등은 1차 항체 회석 농도 1 : 1,000에서도 음성 반응을 나타내었다. 이상의 결과를 종합하면, 간흡충 감염시 숙주의 항체 반응은 충체의 소화-분비 기관에서 유래된 이른바 분비-배설 항원에 의해 가자 강력하게 유발되는 것으로 추측된다.

• 한국식품과학회지
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• 제47권4호
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• pp.534-538
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• 2015

본 연구는 사과 과육을 이용한 잼, 음료, 소스, 통조림 등을 가공하는 과정 중 부수적으로 발생하는 사과 과피의 활용을 위해 70% 에탄올 추출물을 이용하여 생리활성을 측정하였다. 이를 위해 사과 과피 추출물의 총 페놀 및 플라보노이드 함량을 측정하고 항산화 활성과 항염 효과를 평가하였다. 사과 과피 추출물의 총 페놀 함량과 플라보노이드 함량은 각각 $6.8{\pm}0.5mgGAE/g$, 플라보노이드는 $3.3{\pm}0.3mgCE/g$으로 나타났다. 항산화 활성을 평가하기 위해 측정한 DPPH 라디칼 소거능은 추출물의 농도에 의존적으로 증가하여 0.1, 0.5 및 1.0 mg/mL의 농도에서 각각 $18.9{\pm}1.6$, $46.3{\pm}2.3$ 및 $58.1{\pm}3.9%$로 나타났다(p<0.05). 사과 과피 추출물에 의한 염증 관련 효소 활성 억제 효과를 측정한 결과 $sPLA_2$ 활성은 0.5 및 1.0 mg/mL 농도에서 각각 $26.9{\pm}1.4$ 및 $53.5{\pm}2.3%$ 유의적으로 감소하였다(p<0.05). COX-2 활성 억제 효과는 0.1, 0.5 및 1.0 mg/mL의 모든 농도에서 각각 $16.7{\pm}2.6$, $43.6{\pm}3.0$ 및 $64.8{\pm}5.4%$로 유의적이었으나 COX-1 활성은 1.0 mg/mL에서만 13.4% 유의적인 저해효과를 나타냈다(p<0.05). LOX 활성 저해 효과 또한 0.1, 0.5 및 1.0 mg/mL 농도에서 각각 $11.9{\pm}1.3$, $27.4{\pm}2.7$ 및 $44.4{\pm}4.5%$로 나타나 유의적인 항염 효과를 보였다(p<0.05). 본 연구결과는 사과 과피 추출물이 라디칼 소거능을 통한 항산화 효과를 가지고, 다양한 염증 관련 효소의 활성을 억제함으로써 염증 반응을 전반적으로 조절하고 완화시킬 수 있음을 보여주어, 사과를 이용한 제품의 가공 과정에서 부산물로 생기는 과피를 항산화 및 항염 효과를 가지는 건강기능성식품 소재로 이용할 수 있는 가능성을 시사한다.

• 한국동물학회지
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• 제37권4호
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• pp.531-537
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• 1994

Partial internal amino acid sequences of the 15S ATPase from chick skeletal muscle were determined and found to be identical to the corresponding regions of the Mg2+_ATPase from Xenopus laevis oocytes, that is a close homolog of N-ethvlmaleimide-sensitive factor (called NSF) in hamster and Sec18p in yeast, both of which are believed to plaN an essential role in vesicle fusion in secretory process. Thus, the 15S Arpase in chick skeletal muscle maw also belong to a protein family of the "vesicle fusion proteins". Unlike the Mg2'-Afpase with an isoelectric point (pl) of 5.5, however, the 15S Arpase was separated into four spots with pls of 4.9,6.4 and 6.9 upon analysis by twoiimensional gel electrophoresis. In addition, the anti-15S ATPase IgG was found to be capable of interacting with the 265 protease complex upon analysis by immunoprecipitation. Moreover, immunoblot analysis revealed that the anti-155 Arpase IgG recognizes three subunits, ko of which show the same mobilities as the 510-kDa subunit 4 and 48-kDa subunit 7 of the 26S protease complex that are known to contain a highly consented ATP-binding motif. These results surest that a common antigenic site, likely the consensus nucleotide-binding site, exists in the 15S ATPase and the 26S pretense complex and hence both the enzymes maw also be related ATPases.d ATPases.

• Archives of Pharmacal Research
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• 제24권4호
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• pp.307-311
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• 2001

The activation of NF-$\kappa$B induced by kojic Acid, an inhibitor of tyrosinase for biosynthesis of melanin in melanocytes, was investigated in human transfectant HaCaT and SCC-13 cells. These two keratinocyte cell lines transfected with pNF-$\kappa$B-SEAP-NPT plasmid were used to determine the activation of NF-$\kappa$B. Transfectant cells release the secretory alkaline phosphatase (SEAP) as a transcription reporter in response to the NF-$\kappa$B activity and contain the neomycin phosphotransferase (NPT) gene for the dominant selective marker of geneticin resistance. NF-$\kappa$B activation was measured in the SEAP reporter gene assay using a fluorescence detection method. Kojic Acid showed the inhibition of cellular NF-$\kappa$B activity in both human keratinocyte transfectants. It could also downregulate the ultraviolet ray (UVR)-induced activation of NF-$\kappa$B expression in transfectant HaCaT cells. Moreover, the inhibitory activity of kojic Acid in transfectant HaCaT cells was found to be more potent than known antioxidants, e.g., vitamin C and N~acetyl-L-cysteine. These results indicate that kojic Acid is a potential inhibitor of NF-$\kappa$B activation in human keratinocytes, and suggest the hypothesis that NF-$\kappa$B activation may be involved in kojic Acid induced anti-melanogenic effect.

• Journal of Veterinary Science
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• 제23권1호
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• pp.7.1-7.14
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• 2022

Background: Enterotoxigenic Escherichia coli (ETEC) infection is a primary cause of livestock diarrhea. Therefore, effective vaccines are needed to reduce the incidence of ETEC infection. Objectives: Our study aimed to develop a multivalent ETEC vaccine targeting major virulence factors of ETEC, including enterotoxins and fimbriae. Methods: SLS (STa-LTB-STb) recombinant enterotoxin and fimbriae proteins (F4, F5, F6, F18, and F41) were prepared to develop a multivalent vaccine. A total of 65 mice were immunized subcutaneously by vaccines and phosphate-buffered saline (PBS). The levels of specific immunoglobulin G (IgG) and pro-inflammatory cytokines were determined at 0, 7, 14 and 21 days post-vaccination (dpv). A challenge test with a lethal dose of ETEC was performed, and the survival rate of the mice in each group was recorded. Feces and intestine washes were collected to measure the concentrations of secretory immunoglobulin A (sIgA). Results: Anti-SLS and anti-fimbriae-specific IgG in serums of antigen-vaccinated mice were significantly higher than those of the control group. Immunization with the SLS enterotoxin and multivalent vaccine increased interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) concentrations. Compared to diarrheal symptoms and 100% death of mice in the control group, mice inoculated with the multivalent vaccine showed an 80% survival rate without any symptom of diarrhea, while SLS and fimbriae vaccinated groups showed 60 and 70% survival rates, respectively. Conclusions: Both SLS and fimbriae proteins can serve as vaccine antigens, and the combination of these two antigens can elicit stronger immune responses. The results suggest that the multivalent vaccine can be successfully used for preventing ETEC in important livestock.

• 생약학회지
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• 제14권1호
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• pp.30-38
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• 1983

These studies were conducted in an attempt to investigate effects of 'Daeshiho-Tang' on the blood vessels, activities of liver enzyme and excretory action of bile juice in serum of $CCl_4-intoxicated$ rabbits, glucose and total cholesterol levels in serum of alloxan-induced diabetic rabbits. The result of these studies were summarized as follows; 1. Hypotensive ind vaso-dilatating actions due to vascular smooth muscle relaxation were noted in frogs, rabbits and dogs. 2. GOT, GPT and total cholesterol in serum of $CCl_4-intoxicated$ rabbits showed significant depressive effects. 3. Significantly increased biliary secretary effect was noted in $CCl_4-intoxicated$ rabbits. 4. Glucose and total cholesterol levels showed markedly decraese in alloxan-induced diabetic rabbits. Considering the experimental studies results, it is suggested that 'Daeshiho-Tang' has therapeutic efficacy to treat febrile diseses and metabolic diseases.

• 생약학회지
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• 제33권1호통권128호
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• pp.49-52
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• 2002

Amentoflavone, naturally occurring biflavonoid, isolated from the leaves of Ginko biloba, selectively inhibited human seceretory phospholipase $A_2$. This compound potently and irreversibly inhibited human group IIA in a dose dependent manner with an $IC_50$ about $3\;{\mu}M$. Amentoflavone inhibited phospholipase $A_2$ by a noncompetitive manner with the apparent Ki value of $1{\times}10^{-5}M$. In addition, the inhibitory activity of amentoflavone is rather specific against group IIA phospholipase $A_2$ than group IB phospholipase $A_2$. Furthermore, this compound strong inhibit histamine release from $A_{23187}$ treated rat peritoneal mast cells. These results indicate naturally occurring biflavonoid represents a novel anti-inflammatory agent.

• International Journal of Oral Biology
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• 제44권2호
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• pp.62-70
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• 2019

Xylitol is well-known to have an anti-caries effect by inhibiting the replication of cariogenic bacteria. In addition, xylitol enhances saliva secretion. However, the precise molecular mechanism of xylitol on saliva secretion is yet to be elucidated. Thus, in this study, we aimed to investigate the stimulatory effect of xylitol on saliva secretion and to further evaluate the involvement of xylitol in muscarinic type 3 receptor (M3R) signaling. For determining these effects, we measured the saliva flow rate following xylitol treatment in healthy individuals and patients with dry mouth. We further tested the effects of xylitol on M3R signaling in human salivary gland (HSG) cells using real-time quantitative reverse-transcriptase polymerase chain reaction, immunoblotting, and immunostaining. Xylitol candy significantly increased the salivary flow rate and intracellular calcium release in HSG cells via the M3R signaling pathway. In addition, the expressions of M3R and aquaporin 5 were induced by xylitol treatment. Lastly, we investigated the distribution of M3R and aquaporin 5 in HSG cells. Xylitol was found to activate M3R, thereby inducing increases in $Ca^{2+}$ concentration. Stimulation of the muscarinic receptor induced by xylitol activated the internalization of M3R and subsequent trafficking of aquaporin 5. Taken together, these findings suggest a molecular mechanism for secretory effects of xylitol on salivary epithelial cells.

• Parasites, Hosts and Diseases
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• 제31권3호
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• pp.259-268
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• 1993

폐흡충의 분비배설물은 항원성이 높아 폐흡충증의 진단용 항원으로 가치가 있다고 보고되었다. 이 까닭은. 분비배설물에는 숙주내에서 여러 생물학적 반응을 일으키는 물질뿐 아니라 충체의 일부구성 성분 및 여러 효소 등이 포함되어 있기 때문이라 가정할 수 있다. 이 연구는 폐흡충 성충의 분비배설물에서 숙주의 산소라디칼을 분해시키는 효소인 catalase, superoxide dismutasr (SOD), peroxidase 등이 존재하는지를 확인하고 그 활성도를 측정하였으며 그 중 peroxidase를 정제하여 생화학적 특성의 일부 및 항원성을 관찰하였다. 폐흡충 성충 50마리를 $37^{\circ}C$ 부란기에 12시간 배양한 뒤 배양액을 원심분리하고 이를 분비배설 조효소로 사용하였다. 분비배설물에서 catalase, SOD와 peroxidase의 활성도를 측정할 수 있었고 그 비활성도(specific activity)는 각각 11.1, 3.4 및 48.5 이었다. 분비배설물의 peroxidase 비활성도는 충체추출액의 비활성도보다 1.5배 높았다 이 효소를 Sephacryl 5-300 Superfine gel filtration. DEAE-Tnsacryl M anion exchange chromatography로 정제하였다. 정제한 peroxidase의 분자량은 HPLC에서는 19 kDa이었고 SDS-전기영동에서는 16 kDa이었다. 정제한 효소를 전기영동한 후 diaminobenzidine으로 specific staining한 결과 이 효소는 충체추출액의 효소와 같은 영동이동거리를 나타내었다 즉 이 효소는 충체로부터 분비되는 것임을 알 수 있었다. 한편 폐흡충 감염 환자의 혈청과 반응시킨 immunoblot에서 분비배설물의 구성 단백질중 84, 64, 42, 32, 30, 28, 26, 24, 22, 11 및 8 kDa가 항원성을 보인 반면 정제한 peroxidase는 미약한 반응을 보였다 이상의 결과로 폐흡충 peroxidase는 분비배설되어 SOD, catalase와 함께 산소 독성을 제거하는데 작용하고 있으나 패흡충 감염 환자에서 특이 항체 반응을 일으키는데 에는 미약한 작용을 한다는 것을 알 수 있었다.