• 동의생리병리학회지
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• 제20권4호
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• pp.1051-1056
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• 2006

Rehmanniae radix preparata is the root of Rehmanniae glutinosa Liboschitz var. purpurea Makino which has been classified into Scrophulariaceae. Rehmanniae radix preparata has been used for the treatment of diabetes, for the relief of the pain, and for the anti-oxidative action. In this study, the effect of the aqueous extract of Rehmanniae radix preparata on lipopolysaccharide-induced inflammation was investigated by using 3-(4,5-dimethylthiazol- 2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, reverse transcription-polymerase chain reaction (RT-PCR), Western blot, prostaglandin E2 immunoassay, and nitric oxide (NO) detection in mouse BV2 microglial cells. In the present results, the aqueous extract of Rehmanniae radix preparata suppressed prostaglandin E2 (PGE2) synthesis and nitric oxide production by inhibiting the lipopolysaccharide-stimulated expressions of cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) mRNA and protein in mouse BV2 cells. These results show that Rehmanniae radix preparata exerts anti-inflammatory effect probably by suppressing of COX-2 and iNOS expressions.

• Biomolecules & Therapeutics
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• 제20권3호
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• pp.299-305
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• 2012

Endochondral bone formation is the process by which mesenchymal cells condense to become chondrocytes, which ultimately form new bone. The process of chondrogenic differentiation and hypertrophy is critical for bone formation and as such is regulated by many factors. In this study, we aimed to indentify novel factors that regulate chondrogenesis. We investigated the possible role of isopsoralen in induction of chondrogenic differentiation in clonal mouse chondrogenic ATDC5 cells. Isopsoralen treatment stimulated the accumulation of cartilage nodules in a dose-dependent manner. Further, ATDC5 cells treated with isopsoralen were stained more intensely with Alcian blue than control cells, suggesting that isopsoralen increases the synthesis of matrix proteoglycans. Similarly, isopsoralen markedly induced the activation of alkaline phosphatase activity compared with control cells. Isopsoralen enhanced the expressions of chondrogenic marker genes such as collagen II, collagen X, OCN, Smad4 and Sox9 in a time-dependent manner. Furthermore, isopsoralen induced the activation of extracellular signal-regulated kinase (ERK) and p38 MAP kinase, but not that of c-jun N-terminal kinase (JNK). Isopsoralen significantly enhanced the protein expression of BMP-2 in a time-dependent manner. PD98059 and SB 203580, inhibitors of ERK and p38 MAPK, respectively, decreased the number of stained cells treated with isopsoralen. Taken together, these results suggest that isopsoralen mediates a chondromodulating effect by BMP-2 or MAPK signaling pathways, and is therefore a possible therapeutic agent for bone growth disorders.

• Molecules and Cells
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• 제28권5호
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• pp.447-453
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• 2009

The quorum sensing (QS) inhibitors that antagonize TraR, a receptor protein for N-3-oxo-octanoyl-L-homoserine lactones (3-oxo-C8-HSL), a QS signal of Agrobacterium tumefaciens were developed. The structural analogues of 3-oxo-C8-HSL were designed by in silico molecular modeling using SYBYL packages, and synthesized by the solid phase organic synthesis (SPOS) method, where the carboxamide bond of 3-oxo-C8-HSL was replaced with a nicotinamide or a sulfonamide bond to make derivatives of N-nicotinyl-L-homoserine lactones or N-sulfonyl-L-homoserine lactones. The in vivo inhibitory activities of these compounds against QS signaling were assayed using reporter systems and compared with the estimated binding energies from the modeling study. This comparison showed fairly good correlation, suggesting that the in silico interpretation of ligand-receptor structures can be a valuable tool for the pre-design of better competitive inhibitors. In addition, these inhibitors also showed anti-biofilm activities against Pseudomonas aeruginosa.

• 한국식품과학회지
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• 제48권1호
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• pp.72-76
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• 2016

본 연구에서 국립원예특작과학원 인삼특작부 추출물은행에서 분양받은 에탄올 추출물 시료(시료번호: 4)인 선이질풀(Geranium krameri)에 대한 다양한 생리활성을 조사하여 기능성소재 응용가능성을 검토하였다. 선이질풀 추출물은 멜라노마 세포에 대하여 낮은 세포독성을 나타냈다. 세포독성이 거의 없는 농도에서 선이질풀 추출물 처리 시 산화방지 활성($IC_{50}$, $8.72{\mu}g/mL$) 및 B. subtilis, E. coli, C. albicans에 대한 항균활성이 우수하게 나타났으며 타이로시네이즈 활성저해($IC_{50}$, $456.86{\mu}g/mL$) 및 멜라닌 함량 저하($IC_{50}$, $50.35{\mu}g/mL$)를 보여주었다. 또한, B16F10 세포에서 선이질풀 추출물 농도 의존적으로 타이로시네이즈 발현이 억제되었으며, 이는 선이질풀 추출물이 직접적인 타이로시네이즈 활성저해 및 세포 내 타이로시네이즈 발현을 억제시킴으로서 멜라닌 합성을 저해하는 것으로 판단된다. 이와 같은 결과로 미루어볼 때 선이질풀 추출물은 피부미백 소재 등 피부개선 효과를 지닌 기능성 화장품에 활용하기 위한 매우 효과적인 재료가 될 수 있다고 판단된다.

• 한국식품과학회지
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• 제50권3호
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• pp.344-348
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• 2018

본 연구에서 오크라(Abelmoschus esculentus)에 대한 다양한 생리활성을 조사하여 기능성소재 응용가능성을 검토하였다. 오크라 추출물은 멜라노마 세포에 대하여 낮은 세포독성을 나타냈다. 세포독성이 거의 없는 농도에서 오크라 추출물 처리 시, 산화방지활성($ID_{50}$, $5.24{\mu}g/mL$), tyrosinase 활성저해($ID_{50}$, $102.12{\mu}g/mL$) 및 멜라닌 함량 저하($ID_{50}$, $17.85{\mu}g/mL$)를 보여주었다. 오크라 추출물 농도 의존적으로 tyrosinase 발현이 억제되었으며, 이는 오크라 추출물이 직접적인 tyrosinase 활성저해 및 세포 내 tyrosinase 발현을 억제시킴으로써 멜라닌 합성을 저해하는 것으로 판단된다. 또한, 오크라 추출물이 피부장벽 보호 지표물질로서 관련된 단백질인 involucrin 발현을 증가시키는 것을 확인하였다. 이와 같은 결과로 미루어 볼 때 오크라 추출물은 피부미백 소재 등 피부장벽 개선 효과를 지닌 기능성 화장품에 활용하기 위한 매우 효과적인 재료가 될 수 있다고 판단된다.

• 한국식품위생안전성학회지
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• 제30권3호
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• pp.258-264
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• 2015

본 연구는 윈터체리 추출물의 미백 활성을 검증하여 기능성 미백 소재로서의 가능성을 확인하였다. 먼저, DPPH, ABTS, FRAP를 이용한 항산화 활성 검증에서 윈터체리 추출물은 매우 높은 항산화 효과를 보였으며, 티로시나아 제의 활성에도 농도 의존적인 억제 효과를 보여주었다. 마우스 유래 B16-F10 멜라노마 세포를 이용한 멜라닌 생성에서 윈터체리 추출물이 미치는 영향을 확인한 결과, 세포 내에서 생합성 되는 멜라닌의 양이 상당히 감소하고 있음을 확인하였다. 또한, 멜라닌 생성을 더욱 유도하는 ${\alpha}-MSH$를 세포에 처리하였을 때, 윈터체리 추출물은 농도 의존적인 억제 효능을 보여주었다. 이러한 윈터체리 추출물의 멜라닌 생성 억제는 MITF 전사인자의 발현을 억제함으로써 일어나고, 결과적으로 멜라닌 생합성과 관련된 티로시나아제와 Tyrp-1 등의 단백질 발현을 감소시킴으로써 일어나게 됨을 확인하였다. 이러한 결과들로 볼 때, 윈터체리 추출물은 기존의 미백 원료들을 대체할 수 있을 뿐만 아니라 함께 사용할 경우 상승 효과를 낼 수 있는 새로운 미백 소재로 활용될 수 있을 것이다.

• 한국축산식품학회지
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• 제30권3호
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• pp.443-448
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• 2010

유제품, 김치 및 신생아 분변 등에서 유산균을 분리하여 우유 casein을 분해시켜 배양상등액에 존재하는 serine의 함량을 측정함으로써 간접적으로 CPP 생산을 평가하였다. CPP 생산 능력은 OPA방법으로 측정했을 때 E. faecalis A-2 균주가 1.244 mg/mL로 가장 많은 peptide를 생산 하였다. 본 실험에 사용된 유산균의 경우, 세포내 효소와 세포외 효소로 각각 나누어 proteolytic 활성을 평가한 결과 세포내 효소에 의한 proteolytic 활성은 E. faecalis A-6 균주가 가장 높은 것으로 나타났으며, 세포외 효소에 의한 proteolytic 활성은 L. plantarum A-14 균주에서 가장 높게 나타났다. 가용성 칼슘함량은 Leuconostoc mesenteroides A-1(11 mg/dL), E. durans A-16(10.481 mg/dL), 그리고 L. planatarum A-3(10.356 mg/dL) 균주의 경우에는 대조구보다 높은 유리 칼슘함량을 나타내었다. 따라서 이러한 유산균을 이용한다면 소장 내에서 칼슘 흡수를 극대화시킬수 있는 장점이 있어 이들 유산균을 이용한 고칼슘 제품 개발이 가능할 것이다.

• Bulletin of the Korean Chemical Society
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• 제22권12호
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• pp.1361-1365
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• 2001

Recombinant immunotoxins are hybrid cytotoxic proteins designed to selectively kill cancer cells. A divalent immunotoxins, [B3(FabH1)-PE38]2, was constructed by recombining Fab domain of B3 antibody as a cell-targeting domain and Pseudo monas exotoxin A (PE) as a cytotoxic domain. Monoclonal antibody, B3, is the murine antibody (IgG1k) directed against Lewis Y-related carbohydrate antigens, which are abundant on the surface of many carcinomas. Fab fragment of this antibody was used in this study with the modified hinge sequence where last two cysteines out of three were mutated to serine. PE is a 66 kDa bacterial toxin that kills eukaryotic cells by inhibiting protein synthesis with ADP ribosylation of ribosomal elongation factor 2 (EF2). Fc region of B3 antibody was substituted with the truncated form of PE (38 kDa, PE38) on DNA level. [B3(FabH1)-PE38]2 was formed by disulfide bond between cysteines in the modified hinge region of B3(FabH1)-PE38. Each polypeptide for recombinant immunotoxins was overexpressed in Escherichia coli and collected as inclusion bodies. Each inclusion body was solubilized and refolded, and cytotoxic effects were measured. Divalent immunotoxins, [B3(FabH1)-PE38]2, had ID50 values of about 10 ng/mL on A431 cell lines and about 4 ng/mL on CRL1739 cell lines. Control immunotoxins, B3(scFv)-PE40, had ID50 values of about 28 ng/mL on A431 cell lines and about 41 ng/mL on CRL1739 cell lines. Divalent immunotoxins, [B3(FabH1)-PE38]2, had higher cytotoxic effects than B3(scFv)-PE40 control immunotoxins.

• The Korean Journal of Physiology and Pharmacology
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• 제23권6호
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• pp.493-499
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• 2019

Macrophage-associated inflammation is crucial for the pathogenesis of diverse diseases including metabolic disorders. Rhodanthpyrone (Rho) is an active component of Gentiana rhodantha, which has been used in traditional Chinese medicine to treat inflammation. Although synthesis procedures of RhoA and RhoB were reported, the biological effects of the specific compounds have never been explored. In this study, the anti-inflammatory activity and mechanisms of action of RhoA and RhoB were studied in lipopolysaccharide (LPS)-stimulated macrophages. Pretreatment with RhoA and RhoB decreased inducible nitric oxide synthase and cyclooxygenase-2 expressions in RAW 264.7 cells and in thioglycollate-elicited mouse peritoneal macrophages. In addition, it downregulated transcript levels of several inflammatory genes in LPS-stimulated RAW 264.7 cells, including inflammatory cytokines/chemokines (Tnfa, Il6, and Ccl2) and inflammatory mediators (Nos2 and Ptgs2). Macrophage chemotaxis was also inhibited by treatment with the compounds. Mechanistic studies revealed that RhoA and RhoB suppressed the nuclear factor $(NF)-{\kappa}B$ pathway, but not the canonical mitogen activated protein kinase pathway, in LPS-stimulated condition. Moreover, the inhibitory effect of RhoA and RhoB on inflammatory gene expressions was attenuated by treatment with an $NF-{\kappa}B$ inhibitor. Our findings suggest that RhoA and RhoB play an anti-inflammatory role at least in part by suppressing the $NF-{\kappa}B$ pathway during macrophage-mediated inflammation.

• 대한화장품학회지
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• 제28권1호
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• pp.135-149
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• 2002

To date, research on the regulation of melanogenesis has focused on factors which affect tyrosinase, the rate-limiting enzyme in the melanogenic pathway, by searching for chemicals which competitively inhibit tyrosinase function. Many types of tyrosinase inhibitors have been developed, but no satisfactory results have been made clinically until now, To find a new whitening agent, which effectively inhibits melanogenesis, we synthesized several compounds and selected compounds by cell-based assay system. Finally, 3, 4, 5-trimethoxy cinnamaie thymol ester(TCTE, Melasolv) was selected and the effects of TCTE on melanogenesis were investigated. Treatment of mouse-derived melanocyte melan-a cells with TCTE results in a marked down-regulation of tyrosinase activity. 80% decrease of tyrosinase activity occurs with 30uM TCTE treatment for 72 hours without affecting cell growth. The inhibition of tyrosinase activity is dose-dependent and melanin content was also decreased to 40%. From the in vitro tyrosinase assay using cell extract, TCTE does not act as a direct inhibitor of the enzyme. Treatment of melan-a cultures with TCTE blocks the increase in tyrosinase activity by either forskolin, 3-isobutyl-1-methtyl-xanthine. TCTE decreased the expression of tyrosinase, TRP-1 without effects on TRP-2 protein expression through the down regulation of tyrosinase and TRP-1 mRNA. From the results of cAMP immunoassays, intracellular levels of the cyclin nucleotide are unaffected in cells treated with TCTE. The inhibitory effects of melanin synthesis were also shown in reconstitute human epidermis model by topical application. These findings suggest that TCTE can be used for studying the regulation of melanogenesis and depigmenting agent.