Actinobacillys actinomycetemcomitans의 유전자형의 유용성과 세포독성에 관한 연구

조월순,정민호,이상화,황희성 한국생명과학회 2001 생명과학회지 Vol.11 No.1

The purpose of this study was to evaluate the genotypic characterization of Actinobacillus actinomycetemcomitans (A. actinomycetemcomitans) using arbitrarily primed polymerase chain reaction (AP-PCR), to investigate the cytotoxicity of both clinical isolates and standard strains of A. actinomycetemcomitans for the human Jurkat T cells, and to measure the osteoclastogenic cytokines released by Jurkat cells infected with these bacterial strains. The random sequence primer 15 and 16 could distinguish different AP-PCR profiles between clinical isolates of A. actinomycetemcomitans. A. actinomycetemcomitans significantly suppressed Jurkat cell viability in time dependent fashion and the results of DNA fragmentation assay indicated that this bacterial species induced apoptosis in Jurkat cells undergoing apoptosis released the osteoclastogenic cytokine, IL-1$\beta$, IL-6, TNF-$\alpha$. These data support the hypothesis that induction of apoptosis is at least one essential step in A. actinomycetemcomitans induced local immunosuppressive pathway, and that A. actinomycetemcomitans can modulate the immunomodulatory cell population in the periodontal tissue by inducing T cell death through apoptosis, and that apoptosis of local resident T cells may play an active role in bone resorption by releasing osteoclastogenic cytokines, e.g. IL-1$\beta$, IL-6, TNF-$\alpha$.

Resveratrol analogue, HS-1793, inhibits inflammatory mediator release from macrophages by interfering with the TLR4 mediated NF-κB activation

조월순,김성대,정수경,오수정,박문택,이창근,강영록,정민호 한국식품과학회 2022 Food Science and Biotechnology Vol.31 No.4

Resveratrol is known to have anti-inflammatory properties. However, high-dose resveratrol is required for optimal anti-inflammatory effects. HS-1793 is a derivative designed to be metabolically stable and more effective than resveratrol. We tested whether HS-1793 also has anti-in- flammatory activity. HS-1793 effectively inhibited the mRNA and protein expression of lipopolysaccharide (LPS)-induced inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) in macrophages. Therefore, the production of nitric oxide (NO) and prostaglandin E2 (PGE2) was significantly attenuated. In addition, HS-1793 completely suppressed the production of inflammatory cytokines enhanced by LPS treatment along with a decrease in Toll-like receptor 4 (TLR4) expression. At the same time, the expression of myeloid differentiation factor 88 (MyD88), IL-1 receptor-associated kinase 1 (IRAK1), and TNF receptor-associated factor 6 (TRAF6) signaling molecules and the nuclear translocation of nuclear factor kappa B (NF-jB)/p65 were also downregulated. Weconclusively suggest that HS-1793 also exhibits anti-in- flammatory properties by effectively inhibiting TLR4-me- diated NF-jB activation.

굼벵이 유래 밀리타리스 동충하초 열수 추출물의 유전독성평가

조월순(Wol Soon Jo),남병혁(Byung Hyouk Nam),최유진(Yoo Jin Choi),오수정(Su Jung Oh),강은영(Eun-Young Kang),이상호(Sang Ho Lee),정민호(Min Ho Jeong) 한국독성학회 2007 Toxicological Research Vol.23 No.3

Water extract of Cordyceps militaris grown upon Protuetja dreujtarsis (CMPD) was examined for the genetic toxicity-bacterial mutagenicity, chromosome aberration, and micronucleus formation. For mutagenicity assay, bacterial reversion test with Salmonella typhimurium TA98, TA100, TA1535, TA 1537, and E. coli WP2uvrA were performed. The extract at the concentrations of 50~5,000 ㎍/plate did not induce mutagenicity at all. Chromosome aberration test was performed by using Chinese lung (CHL) cells. There was no significant chromosome aberration in CHL cells with S-9 mixture at the concentrations of 312.5~1,250 ㎍/㎖ of the extract and without S-9 mixture at the concentrations of 1.2~19.5 ㎍/㎖ of the extract. For micronucleus test, ICR mice were treated with the extract at the dose of 0.5, 1, and 2 g/㎏. The frequencies of the micronucleated polychromatic erythrocytes (MNPCE) in bone marrow preparations of the extract-treated group were not increased compared to the untreated control group. Taken together, our results show that water extract of CMPD did not induce any harmful genotoxicity.

굼벵이 유래 밀리타리스 동충하초 열수추출물의 간기능개선 효과 및 단회독성 평가

조월순(Wol-Soon Jo),남병혁(Byung-Hyouk Nam),오수정(Su-Jung Oh),최유진(Yoo-Jin Choi),강은영(Eun-Young Kang),홍숙희(Sook-Hee Hong),이상호(Sang-Ho Lee),정민호(Min-Ho Jeong) 한국식품과학회 2008 한국식품과학회지 Vol.40 No.1

굼벵이 유래 밀리타리스 동충하초 열수추출물(CMPD extract)의 안전성 자료를 확보하기 위하여 단회경구투여 독성시험을 실시하였으며, CCl₄의 경구투여로 유도된 간손상 실험동물모델로부터 시험물질(CMPD extract)의 간보호효과를 확인하였다. 단회경구투여 독성시험결과, 시험물질(CMPD extract)의 최고농도(2,000 mg/kg body weight)에서 독성을 나타내지 않아 LD<SUB>50</SUB> 값을 그 이상으로 결정하였다. 또한 독성물질(CCl₄)로부터 간손상이 유발된 SD rat에 시험물질(CMPD extract)을 투여한 후 혈청으로부터 간손상과 관련한 지표물질인 GOT, GPT, TG, TC, LDL 및 HDL 활성도를 측정하였으며, 이와 함께 병리조직학적 소견을 확인하였다. 혈청 GOT는 손상군(G2)에 비해 G4, G5 시험물질투여군에서 유의한 감소를 나타내었으며, GPT의 경우 고용량 시험물질 투여군(G4)에서 유의한 감소를 나타내었다(p < 0.05). 또한 LDL 및 HDL 활성도는 손상군(G2)에 비해 유의하지는 않지만 시험 물질투여군(G4, G5, G6)에서 어느 정도 회복기미를 보였다. 병리조직학적 소견에서도 손상군(G2)의 경우 심각한 세포독성을 보였으나, 시험물질 고용량 투여군(G4)에서는 손상된 세포가 감소하였음을 확인하였다. 이상의 결과들은 굼벵이 유래 밀리타리스 동충하초 열수추출물(CMPD extract)이 CCl₄ 투여에 의해 유발된 급성간 손상에 대하여 간조직의 보호와 간세포의 기능유지에 유효한 물질임을 제시 하고 있다. This study was designed to evaluate the single dose toxicity and the protective effect of water extract of Cordyceps militaris grown upon Protaetia dreujtarsis (CMPD extract) on liver damage on carbon tetrachloride (CCl₄)-induced acute hepatotoxicity in Sprague-Dawley (SD) rats. The CMPD extract was once administered orally to both sexes of rats at dose of 2,000, 1,000 and 500 mg/kg body weight, the recommended maximum limit dose for acute toxicity. Neither significant toxic signs nor death was observed during the observation period. These results indicate that LD<SUB>50</SUB> (lethal dose of 50%) of CMPD extract is greater than 2,000 mg/kg body weight in SD rats. To investigate also the effect of hepatoprotection of CMPD extract, SD rats were orally treated with CMPD extract (50, 25 and 12.5 mg/kg body weight) or silymarin (25 mg/kg body weight) before and after administration of CCl₄ (2 mL/kg body weight, 20% CCl₄ in olive oil). Treatment with CMPD extract or silymarin could decrease the GPT (glutamic-pyruvic transaminase) and GOT (glutamic-oxaloacetic transaminase) levels in serum when compared with CCl₄-treated group. Therefore, the results of this study show that CMPD extract can be proposed to protect the liver against CCl₄-induced hepatic damage in rats.

In Vitro Genotoxicity Assessment of a Novel Resveratrol Analogue, HS-1793

정민호,조월순,양광모,이창근,정동혁,박유수,최유진,김중선,오수정,정수경 한국독성학회 2014 Toxicological Research Vol.30 No.3

Resveratrol has received considerable attention as a polyphenol with various biological effects such as anti-inflammatory, anti-oxidant, anti-mutagenic, anti-carcinogenic, and cardioprotective properties. As part of the overall safety assessment of HS-1793, a novel resveratrol analogue free from the restriction of metabolic instability and the high dose requirement of resveratrol, we assessed genotoxicity in three in vitro assays: a bacterial mutation assay, a comet assay, and a chromosomal aberration assay. In the bacterial reverse mutation assay, HS-1793 did not increase revertant colony numbers in S. typhimurium strains (TA98, TA100, TA1535 and TA1537) or an E. coli strain (WP2 uvrA) regardless of metabolic activation. HS-1793 showed no evidence of genotoxic activity such as DNA damage on L5178Y Tk+/- mouse lymphoma cells with or without the S9 mix in the in vitro comet assay. No statistically significant differences in the incidence of chromosomal aberrations following HS-1793 treatment was observed on Chinese hamster lung cells exposed with or without the S9 mix. These results provide additional evidence that HS-1793 is non-genotoxic at the dose tested in three standard tests and further supports the generally recognized as safe determination of HS-1793 during early drug development.

항종양 면역반응 유도를 위한 수지상세포의 최적 활성화 조건

남병혁,조월순,이기원,오수정,강은영,최유진,도은주,홍숙희,임영진,김기욱,정민호,Nam, Byung-Hyouk,Jo, Wool-Soon,Lee, Ki-Won,Oh, Su-Jung,Kang, Eun-Young,Choi, Yu-Jin,Do, Eun-Ju,Hong, Sook-Hee,Lim, Young-Jin,Kim, Ki-Uk,Jeong, Min-Ho 한국생명과학회 2006 생명과학회지 Vol.16 No.6

수지상세포는 종양면역에서 필수적인 강력한 CTL 반응을 개시할 수 있는 유일한 세포이다 . 특히 외인성 종양항원에 대한 CTL 반응 유도는 활성화된 수지상세포의 IL-12 분비를 통한 CD4+ helper T세포의 cross-priming을 필요로 한다. 그러나 최근에 활성화된 수지상세포는 $Th_1$ 면역반응을 유도하지만 활성화 시간이 경과함에 따라 오히려 $Th_2$반응을 유도 할 수 있다. 따라서 본 연구에서는 OVA를 종양항원 모델로 설정하여 종양특이적인 CTL 반응을 형성하기 위한 최적의 수지상세포 활성화 조건을 조사하였다. 마우스 골수세포에 서 수지상세포로의 분화는 항원제시 기능을 위한 표면분자의 발현 측면에서 볼 때 배양 6일-7일 정도가 적합하였다. 수지상세포의 IL-12 생성능은 배양 6일 이상, OVA 항원 탑재 8시간 이상의 경우에 연이은 LPS 성숙자극으로 오히려 감소하는 경향을 보였다. 즉 배양 6일의 수지상세포에 OVA 항원 탑재를 8시간 수행한 경우(8-h DC)가 in vitro에서의 IL-12생성능, ex vivo에서의 세포내 $IFN-{\gamma}$를 발현하는 CD8+ T세포의 증가 및 OVA 특이적인 세포독성효과 등에서 가장 좋은 결과를 보였다. 또한 in vivo에서 종양 치료 및 예방효과에서도 8-h DC로 면역한 경우에 가장 우수한 종양형성 억제 효과와 생존기간 연장효과를 보였다. 현재 대부분의 수지상 세포를 이용한 항종양 백신에서 항원 탑재반응을 24시간 동안 수행하고 있으나, 본 실험의 결과로 볼 때, 8시간의 in vitro 항원 탑재가 보다 효과적인 종양특이적 CTL 반응과 항종양 면역반응을 유도함을 알 수 있다. 결론적으로 본 연구를 통하여 8시간 이상의 항원접촉은 수지상세포의 기능적 활성능력을 오히려 고갈시킬 수 있음을 제시한다. Dendritic cells (DCs) are the only antigen presenting cells (APCs) capable of initiating immune responses, which is crucial for priming the specific cytotoxic T lymphocyte (CTL) response and tumor immunity. Upon activation by DCs, CD4+ helper T cells can cross-prime CD8+ CTLs via IL-12. However, recently activated DCs were described to prime in vitro strong T helper cell type 1 $(Th_1)$ responses, whereas at later time points, they preferentially prime $Th_2$ cells. Therfore, we examined in this study the optimum kinetic state of DCs activation impacted on in vivo priming of tumor-specific CTLs by using ovalbumin (OVA) tumor antigen model. Bone-marrow-derived DCs showed an appropriate expression of surface MHC and costimulatory molecules after 6 or 7-day differentiation. The 6-day differentiated DCs pulsed with OVA antigen for 8 h (8-h DC) and followed by restimulation with LPS for 24 h maintained high interleukin (IL)-12 production potential, accompanying the decreased level in their secretion by delayed re-exposure time to LPS. Furthermore, immunization with 8-h DC induced higher intracellular $interferon(IFN)-{\gamma}+/CD8+T$ cells and elicited more powerful cytotoxicity of splenocytes to EG7 cells, a clone of EL4 cells transfected with an OVA cDNA, than immunization with 24-h DC. In the animal study for the evaluation of therapeutic or protective antitumor immunity, immunization with 8-h DC induced an effective antitumor immunity against tumor of EG7 cells and completely protected mice from tumor formation and prolonged survival, respectively. The most commonly used and clinically applied DC-based vaccine is based on in vitro antigen loading for 24 h. However, our data indicated that antigen stimulation over 8 h decreased antitumor immunity with functional exhaustion of DCs, and that the 8-h DC would be an optimum activation state impacted on in vivo priming of tumor-specific CTLs and subsequently lead to induction of strong antitumor immunity.

고지혈증을 가진 뇌경색 환자에서 Statin에 의한 혈소판의 CD63과 CD40L의 발현의 억제

이경미,조월순,정민호,임영진,박귀선,차재관 대한신경과학회 2004 대한신경과학회지 Vol.22 No.5

Background: Statin (HMG-coA-reductase inhibitor) has been known to protect vessels from atherothrombosis through various mechanisms. In this study, we evaluated the effects of statin on reducing the platelet expressions of CD63 and CD40 ligand (CD40L) in subjects with atherosclerotic ischemic stroke. Methods: Twenty-one patients (17 men, 4 women; mean age 59.0±10.2 years) with atherosclerotic ischemic stroke were recruited. They took simvastatin 20 mg per day for 90 days and discontinued for another 90 days. We studied the changes of platelet expressions of CD63 and CD40L in all the patients after the use and discontinuance of simvastatin using whole blood flow cytometry. Results: After taking simvastatin 20mg for 90 days, the serum concentrations of LDL cholesterol decreased significantly (96.4±31.4 mg/dL, p<0.001) compared with those at the baseline (158.8±25.0 mg/dL). The platelet CD63 and C40L expressions were also significantly reduced by treatment of simvastatin 20 mg for 12 weeks (p<0.05). However, the effects of statin on CD63 and CD40L expressions disappeared after 12 weeks of cessation. Furthermore, changes of expressions of CD63 and CD40L by statin did not correlate with its cholesterol lowering effect (r=-0.311, p=0.386). Conclusions: This study demonstrates that the use of statin may be a helpful strategy to regulate the platelet activation in patients with atherosclerotic ischemic stroke.

Low dose rate radiation regulates M2-like macrophages in an allergic airway inflammation mouse model

강소희,조월순,이창근,배민지,문창종,정소희,손영훈,이해준,김중선 대한방사선방어학회 2022 대한방사선방어학회 학술발표회 논문요약집 Vol.2022 No.04

Chronic low-dose-rate radiation exposure leads to testicular damage

강소희,배민지,조월순,문창종,손영훈,이해준,이창근,김중선 대한방사선방어학회 2022 대한방사선방어학회 학술발표회 논문요약집 Vol.2022 No.04

방사성의약품 99mTc를 이용한 감마분광법 유효성평가

이지은,김효진,계용욱,장희진,조월순,이창근,강영록 대한방사선방어학회 2023 대한방사선방어학회 학술발표회 논문요약집 Vol.2023 No.4