• 한국식품과학회지
• /
• 제39권6호
• /
• pp.708-714
• /
• 2007

식품 소재의 국내산 약용식물인 163가지 천연물에서 메탄올로 추출한 시료를 이용하여 in vivo에서 10, 100, 500, $1000\;{\mu}g/mL$의 4가지 농도에서 대식세포 면역 활성을 측정하였다. 그 결과, 42개의 시료에서 면역증진반응을 보였으며, 그 중 20개의 시료는 음성대조군에 대하여 20% 이상 면역 활성을 증진시키는 것으로 측정되었다. 이중 2가지 농도에서 면역증진반응을 보이는 시료는 총 19개, 3가지 농도에서 면역증진효과를 나타내는 시료는 총 3개[골파(Allium schoenoprasmum), 두릅(Aralia elata), 매생이(Capsosiphon fulvescens)]였으며, 특히 마(Dioscorea batatas)는 각 농도에서 활성을 나타내었을 뿐 아니라 양성대조군과 비슷하거나 높은 활성을 나타내어 면역증진활성이 매우 우수한 것으로 나타났다.

• 대한의생명과학회지
• /
• 제10권2호
• /
• pp.65-73
• /
• 2004

I report that single-stranded antisense as a part of large circular (LC-) genomic DNA of recombinant M13 phage exhibits enhanced stability, sequence specific antisense activity, and no need for target site search. A cDNA fragment (708 bp) of rat TNF-$\alpha$ was inserted into a phagemid vector, and TNF-$\alpha$ antisense molecules (TNF$\alpha$-LCAS) were produced as single-stranded circular DNA. When introduced into a rat monocyte/macrophage cell line, WRT7/P2, TNF$\alpha$-LCAS was able to ablate LPS-induced TNF-$\alpha$ mRNA to completion. The antisense effect of TNF$\alpha$-LCAS was shown to be sequence-specific because expressions of three control genes ($\beta$-actin, GAPDH and IL-1$\beta$) were not significantly altered by the antisense treatment. Further, TNF$\alpha$-LCAS was found to be highly efficacious as only 0.1 $\mu$g (0.24 nM) of TNF$\alpha$-LCAS was sufficient to block TNF-$\alpha$ expression in 1$\times10^5$ WRT7/P2 cells. I have also observed specific antisense activity in reduction of NF-$\kappa$B gene expression. The results suggest that an antisense sequence as a part of single-stranded circular genomic DNA has a specific antisense activity.

• 한국식품과학회지
• /
• 제53권3호
• /
• pp.344-347
• /
• 2021

본 연구에서는 chaparral 70% 에탄올 추출물을 0.01-100 ㎍/mL의 농도로 HepG2 간세포에 처리하여 세포 사멸, 염증성 사이토카인 분비, 세포 내 지방 축적을 측정하는 in vitro 실험을 통해 chaparral의 간독성 기전을 조사하였다. Chaparral 추출물 처리에 의해 1-100 ㎍/mL의 농도에서 세포 사멸이 관찰되었으며, 염증성 사이토카인인 IL-8과 M-CSF의 분비 및 지방 축적은 10배 더 낮은 농도부터인 0.1-100 ㎍/mL에서 유의적으로 증가하였다(p<0.05). 본 연구결과에서 염증성 사이토카인 분비 및 지방 축적을 통한 간세포 염증은 chaparral 추출물에 의해 유발되는 간독성의 한 형태로 나타났다. 또한 간염 형태의 간독성은 세포 사멸이라는 심각한 독성을 야기시키는 농도보다 낮은 농도에서 발현되어 저농도의 chaparral 섭취에 의해 간염과 같은 간독성이 초래될 수 있음을 시사한다.

• Journal of Applied Biological Chemistry
• /
• 제57권3호
• /
• pp.205-210
• /
• 2014

Persicaria orientalis (L.) Spach (Po)와 Potentilla fragarioides var. major Maxim (Pf)의 추출물로 reactive oxygen species (ROS)와 같은 free radical의 억제를 통한 항염증 활성을 살펴보았다. 또한, Po와 Pf로 LPS를 처리한 murine macrophage RAW 264.7 세포에서 발생한 nitric oxide의 저해와 NF-${\kappa}B$의 핵으로 translocation의 저해를 살펴보았다. 3-morpholinosydnonimine hydrochloride (SIN-1) $50{\mu}M$에 의해 유도된 ROS의 Po에 의한 50% 저해값 ($IC_{50}$)은 $23.35{\pm}1.27mg/mL$, Pf에 의한 $IC_{50}$은 $8.46{\pm}1.22mg/mL$이었다. 또한, SIN-1 $50{\mu}M$에 의해 유도된 peroxynitrite의 Po에 의한 $IC_{50}$은 $2.19{\pm}0.04mg/mL$, Pf에 의한 $IC_{50}$은 $0.80{\pm}0.02mg/mL$이었다. LPS 1 mg/mL을 처리한 RAW 264.7 세포에서 Nitric oxide는 증가하였으나 Po와 Pf 추출물을 처리한 그룹에서 농도의존적, 유의적으로 감소하였다. Po 추출물을 처리한 그룹의 Nitric oxide 생성량은 $13.34{\pm}0.67{\mu}M$, Pf 추출물을 처리한 그룹의 Nitric oxide 생성량은 $11.45{\pm}0.57{\mu}M$이었다. 또한, Po와 Pf 추출물은 LPS를 처리한 RAW 264.7 세포에서 NF-${\kappa}B$의 핵으로의 translocation을 저해하였다. 그러므로, Po와 Pf는 항염증 소재로서의 가능성이 충분하다고 사료된다.

• 대한본초학회지
• /
• 제37권6호
• /
• pp.19-28
• /
• 2022

Objectives : To develop natural ingredients that help prevent or treat anti-inflammatory-related diseases and use themas basic data, we investigated anti-inflammatory activity of Cynanchi Atrati Radix Et Rhizoma water extracts(CWE) in lipopolysaccharide(LPS)-induced murine macrophage cell line, RAW 264.7 cells. Methods : The cell viabilities were evaluated with RAW 264.7 cells. The production of nitric oxide(NO), prostaglandin E2(PGE2), pro-inflammatory cytokines such tumor necrotic factor(TNF)-α and interleukin(IL)-6 were assessed in LPS-induced RAW 264.7 cell treated with CWE. Furthermore, the protein expression of inducible nitric oxide synthase (iNOS), cyclooxygenase-2(COX-2), and mitogen-activated protein kinase(MAPK) were assessed by western blotting. Results : In RAW 264.7 cell, the cell viability by CWE treatment was more than 98.4% at a concentration of 100-400 ㎍/mL. At a concentration of 800 ug/ml of CWE, the cell viability was as low as 86%. At doses of 100, 200 and 400 ㎍/mL, CWE inhibited the production of NO, PGE2, TNF-𝛼 and IL-6 in a dose-dependent manner and also decreased the expression of iNOS and COX-2 from LPS-induced RAW 264.7 cells. In addition, CWE significantly inhibited the MAPK pathway including decreased the phosphorylation of the p38, c-Jun N-terminal kinase(JNK) and extracellular signal-regulated kinase(ERK1/2). Conclusions : Our study provides evidence that CWE inhibits the production of main pro-inflammatory molecules in LPS-induced RAW 264.7 cells via expression of p38, JNK, and ERK1/2 MAPK signaling pathways. Therefore, CWE is expected to be widely used as a natural ingredient for anti-inflammatory functional foods or pharmaceuticals in the future.

• 대한의생명과학회지
• /
• 제23권4호
• /
• pp.327-332
• /
• 2017

A2B adenosine receptor (A2BAR) is known to be a regulator of bone homeostasis, but the regulatory mechanism of A2BAR on the osteoclast proliferation are poorly explored. Recently, we have shown that stimulation with BAY 60-6583, a specific agonist of A2BAR, significantly reduced macrophage-colony stimulating factor (M-CSF)-induced osteoclast proliferation by inducing cell cycle arrest at G1 phase and increasing the apoptosis of osteoclasts. The objective of this study was to investigate the regulatory mechanisms of cell cycle and apoptosis by A2BAR stimulation. The expression of A2BAR and M-CSF receptor, c-Fms, was not changed by A2BAR stimulation whereas M-CSF effectively induced c-Fms expression during osteoclast proliferation. Interestingly, A2BAR stimulation remarkably increased the expression of $p27^{Kip-1}$, a cell cycle inhibitor, but the expression of Cyclin D1 and cdk4 was not affected. In addition, while BAY 60-6583 treatment reduced the expression of Bcl2, an anti-apoptotic oncogene, it failed to regulate the expression of Bax, a pro-apoptotic marker. Taken together, these results imply that the increase of $p27^{Kip-1}$ inducing cell cycle arrest at G1 phase and the decrease of Bcl2 inducing anti-apoptotic response by A2BAR stimulation contribute to the down-regulation of osteoclast proliferation.

• Journal of Periodontal and Implant Science
• /
• 제32권4호
• /
• pp.865-873
• /
• 2002

The primary cause of tooth loss after 30 years of age is periodontal disease. Destruction of alveolar bone by periodontal disease is done by bone resorbing activity of osteoclasts. Understanding differentiation and activation mechanism of osteoclasts is essential for controling periodontal disease. The purpose of this study is to identify the possible effects of Vitamin D and cytokines affecting osteoclasts and its precursor cells. Four to six week-old mice were killed and humerus, radius, tibia and femur were removed aseptically and washed two times with Hank's solution containing penicillin-streptomycin and then soft tissue were removed. Bone marrow cells were collected by 22 gauge needle. Cells were cultured in Hank's solution containing 1 mg/ml type II collagenase, 0.05% trypsin, 41mM EDTA. Supernatant solution was removed 5 times after 15 minutes of digestion with above mentioned enzyme solution, and remained bone particles were maintained in alpha-MEM for 15 minutes and $4^{\circ}C$ temperature. Bone particles were agitated for 1 minute and supernatant solution containing osteoclast precursor cells were filtrated with cell stainer. These separated osteoclast precursor cells were dispensed with 100-mm culture dish by $1{\times}10^7$ cells unit and cultured in ${\alpha}$- MEM containing 20 ng/ml recombinant human M-CSF, 30 ng/ml recombinant human soluble osteoclast differentiation factor and 10% fetal calf serum for 2 and 7 days. Total RNA of osteoclast precursor cells were extracted using RNeasy kit. One ${\mu}g$ of total RNA was reverse transcribed in $42^{\circ}C$ for 30 minutes using SuperScriptII reverse transcriptase. Expression of transcribed receptors of each hormone and cytokine were traced with 1 ${\mu}l$ of cDNA solution by PCR amplification. Vitamin D receptor WAS found in cells cultured for 7 days. TNF-${\alpha}$ receptor was found in cells cultured for 2 days and amount of receptors were increased by 7 days. IL-1 type I receptor was not found in cells cultured 2 and 7 days. But, IL-1 receptor type II was found in cells cultured for 2 days. TGF-${\alpha},{\beta}$type I receptor was found in cells cultured 2 and 7 days, and amount of receptors were increased by 7 days of culture. These results implies Vitamin D and cytokines can affect osteoclasts directly, and affecting period in differentiation cycle of osteoclasts is different by Vitamin D and cytokines.

• Asian Pacific Journal of Cancer Prevention
• /
• 제16권7호
• /
• pp.3043-3050
• /
• 2015

The tumor microenvironment (TME) includes numerous non-neoplastic cells such as leukocytes and fibroblasts that surround the neoplasm and influence its growth. Tumor-associated macrophages (TAMs) and cancerassociated fibroblasts (CAFs) are documented as key players in facilitating cancer appearance and progression. Alteration of the macrophage (CD68, CD163) and fibroblast (${\alpha}-SMA$, FSP-1) cells in Opisthorchis viverrini (Ov) -induced cholangiocarcinoma (CCA) was here assessed using liver tissues from an established hamster model and from 43 human cases using immunohistochemistry. We further investigated whether M2-activated TAMs influence CCA cell migration ability by wound healing assay and Western blot analysis. Macrophages and fibroblasts change their phenotypes to M2-TAMs (CD68+, CD163+) and CAFs (${\alpha}-SMA+$, FSP-1+), respectively in the early stages of carcinogenesis. Interestingly, a high density of the M2-TAMs CCA in patients is significantly associated with the presence of extrahepatic metastases (p=0.021). Similarly, CD163+ CCA cells are correlated with metastases (p=0.002), and they may be representative of an epithelial-to-mesenchymal transition (EMT) with increased metastatic activity. We further showed that M2-TAM conditioned medium can induce CCA cell migration as well as increase N-cadherin expression (mesenchymal marker). The present work revealed that significant TME changes occur at an early stage of Ov-induced carcinogenesis and that M2-TAMs are key factors contributing to CCA metastasis, possibly via EMT processes.

• Bulletin of the Korean Chemical Society
• /
• 제30권8호
• /
• pp.1839-1844
• /
• 2009

A 20-residue hybrid peptide CA(1-8)-MA(1-12) (CAMA) incorporating residues 1-8 of cecropin A (CA) and residues 1-12 of magainin 2 (MA) has high antimicrobial activity without toxicity. To investigate the effects of the total positive charges of CAMA on the antibacterial activity and toxicity, a hybrid peptide analogue (CAMA-syn) was designed with substitutions of $Ile^{10}\;and\;Ser^{16}$ with Lys. According to CD spectra, structure of CAMA-syn with increase of cationicity was very similar to that of CAMA in DPC micelle. CAMA-syn showed antimicrobial activity similar with CAMA while CAMA-syn has no hemolytic activity and much lower cytotoxicity against RAW 264.7 macrophage cells than CAMA. Also, CAMA and CAMA-syn significantly inhibited NO production by LPSstimulated RAW264.7 macrophage at 10.0∼20.0 $\mu$M. CAMA-syn displayed salt resistance on antimicrobial activity against Escherichia coli at the physiological concentrations of $CaCl_2\;and\;MgCl_2$. The combination studies of peptides and antibiotics showed that CAMA-syn has synergistic effects with synthetic compound and flavonoid against Enterococcus faecalis and VREF. CAMA-syn can be a good candidate for the development of new antibiotics with potent antibacterial and synergistic activity but without cytotoxicity.

• 대한약학회:학술대회논문집
• /
• 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2-2
• /
• pp.194.4-195
• /
• 2003

Two phenolic flavonoids were isolated from the traditional medicine of Eastern Asia, Caesalpinia sappan L (Leguminosae). Brazilin (1) showed a significant inhibitory activity against inducible Nitric Oxide Synthase (iNOS) in lipopolysaccharides (LPS)-induced macrophage RAW 264.7 cells with an IC$\sub$50/ value of 1.68 mg/ml, which is more potent than the positive control, L-N$\^$6/-(1-iminoethyl)lysine (IC$\sub$50/ 3.49 mM). On the other hand, caesalpine J (2) was found to be inactive in the present iNOS assay system despite of their structural similarities. (omitted)