• Journal of Periodontal and Implant Science
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• v.27 no.4
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• pp.805-818
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• 1997

Immune responses of periodontal tissue may be regulated by products of sensory afferent nerve endings such as neuropeptides. Substance P(SP), a tachykinin neuropeptide, has been previously reported to stimulate the activities of T lymphocyte. Therefore, I examined the role of SP in IL-2 production and cell proliferation by using a homogeneous line of T lymphocytes(Jurkat and HuT78). Cell proliferation rate was determined by [$^3H$]-thymidine incorporation test, and IL-2 was quantitated by the growth rate of CD4+ IL-2-dependent T lymphocyte line CTLL-2. SP stimulated cell proliferation of T lymphocytes at the concentration of $10^{-12}$ and $10^{-8}$M in a biphasic bell-shape dose-dependent manner. However, SP alone did not induce IL-2 release at the concentration range of $10^{-6}$ to $10^{-14}$M. The upregulation of IL-2 release was observed when $10^{-12}$M SP was applied together with mitogens such as Con A or PHA+PMA on T cell lines, especially on Jurkat. Con A or PHA+PMA demonstrated to increase the rate of cell proliferation of Jurkat, which had shown to produce much amount of IL-2 indicating that mitogen-induced cell proliferation might be partially influenced by released IL-2. It was concluded that regulatory effects of SP on the immune/inflammatory response could be mediated through the costimulatory upregulation of IL-2 production and increase of cell proliferation of T lymphocyte.

• Restorative Dentistry and Endodontics
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• v.27 no.5
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• pp.473-478
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• 2002

근관내 spirochetes의 존재유무가 명확하게 밝혀져 있지 않았으나 최근 PCR을 사용한 연구에서 Treponema denticola균주가 감염근관의 50% 이상의 경우에서 발견됨에 따라 이 세균이 치수 및 치근단 질환에 관여하는지에 대한 관심 이 높아졌다. 하지만 그 정확한 기전은 아직 밝혀져 있지 않다. 이와 관련하여 Shenker등이 T. denticola의 sonicated extract에서 순수분리된 단백질 (SIP)이 임파구 proliferation을 방해함을 보고한바 있다. 따라서 본 연구의 목적은 면역억제단백질 SIP이 어떤 기전에 의해서 임파구증식을 억제하는지를 밝히는 데 있다. 건강한 혈액 공여자로부터 추출해낸 T세포에 PHA (phytohemagglutinin)로 증식자극을 주게되는데 이 과정에서 SIP을 처리하거나 처리하지 않은 경우를 비교하여 세포주기 진행과정을 유세포분석기 (Becton-Dickinson FACS$^{tarplus}$) 를 통하여 평가하였다. 실험결과 세단계의 chromatography과정을 통해 순수정제된 SIP은 50kDa와 56kDa의 두가지 polypeptide로 구성되어 있고 0.25$\mu\textrm{g}$으로 처리된 T 임파구는 42.5%의 [$^3$H]thymidine incorporation 억제가 그리고, 0.5$\mu\textrm{g}$으로 처리한 경우는 75.1%의 억제가 일어나 dose-dependent한 양상이 나타났다. Propidium iodide와 유세포 분석기를 사용하여 세포주기를 분석한 결과 medium으로만 처리한 경우 97%이상의 임파구는 G$_0$/G$_1$ phase에 머물러 있었으나 PHA자극을 받은 경우 G$_0$/G$_1$ phase에서 58%, S phase에서 34.6%, G$_2$/M phase에서 7.4%로 분포되어 나타났다. SIP으로 전처리한 경우 세포 증식이 감소하여 0.25$\mu\textrm{g}$을 첨가한 경우 75.1%가 G$_0$/G$_1$ phase에 머물러 있었고 더 강한 농도의 0.5$\mu\textrm{g}$을 첨가한 경우는 87.7%가 G$_0$/G$_1$ phase에서 S phase로 진행되지 않고 머물러있었다. 따라서 SIP으로 전처리된 T 임파구는 그 증식이 G$_0$/G$_1$ phase에서 차단된 것으로 보인다. 이러한 면역억제현상이 in vitro 상태뿐 아니라 in vivo에서도 진행된다면 spirochete가 치수 및 치근단 질환의 병인론에 연관된 면역반응저하기전에 중요한 역할을 하는 것으로 추론할 수 있다.

• Animal cells and systems
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• v.1 no.1
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• pp.151-155
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• 1997

Promoter of the Drosophila proliferating cell nuclear antigen (PCNA) gene contains DRE (Drosophila DNA replication-related element) required for the high level expression of replication-related genes. Recently, we found that promoter region of the D-raf (a Drosophila homolog of the human c-raf-1) contains two sequences homologous to the DRE and demonstrated the DRE/DREF (DRE-binding factor) involvement in regulation of the D-raf gene. In this study, using ursolic acid (UA), a pentacyclic triterpene acid reported to possess antitumor activities, we examined effects of UA on proliferation of the Drosophila cultured Kc cells and on expression of the PCNA and D-raf genes. UA showed an inhibitory effect on proliferation of the Kc cells in a concentration-dependent manner in DNA content assays and [3H]thymidine incorporation assays. The IC50 value of anti-proliferative effects of UA in DNA content assays was about 7.5uM. UA showed inhibitory effects on expression of the PCNA as well as on that of the D-raf, which were examined with the reporter plasmic p5'-168DPCNACAT or p5'-878DrafCAT, respectively. The results obtained in the present study suggest that expression of the PCNA and D-raf genes is coordinately regulated in at least UA-treated Kc cells and that down-regulation of expression of the PCNA and D-raf genes might be related with the antitumor activities of UA.

• YAKHAK HOEJI
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• v.55 no.2
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• pp.116-120
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• 2011

Abnormal proliferation of vascular smooth muscle cells (VSMCs) plays an important role in the development and progression of proliferative cardiovascular diseases, including hypertension and atherosclerosis. To find antiproliferative agent (A)-8 had inhibitory effect on VSMCs proliferation. Therefore, we examined the antiproliferative effect of A-8, a newly synthesized obovatol derivative. To investigate the antiproliferative effect of A-8, we examined cell counting and [$^3H$]-thymidine incorporation assays. The pre-incubation of A-8 (1~4 ${\mu}M$) significantly inhibited proliferation and DNA synthesis of 5% fetal bovine serum (FBS)-stimulated rat aortic VSMCs in concentration-dependent manner. Whereas, A-8 did not show any cytotoxicity in rat aortic VSMCs in this experimental condition by WST-1 assay. In addition, A-8 significantly inhibited 5% FBS-induced cell cycle progression in rat aortic VSMCs. These results show that A-8 may be developed as a potential antiproliferative agent for treatment of angioplasty restenosis and atherosclerosis. Furthermore, A-8 should be examined for further clinical application either as a single agent or in combination with other angioplasty restenosis or atherosclerosis agents.

• The Korean Journal of Zoology
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• v.25 no.2
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• pp.55-62
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• 1982

DNA synthesis, unscheduled DNA synthesis, excision of pyrimidine dimers and phtoreactivation were determined in UV-irradiated differentiating muscle cells at various times of primary culture of 12 day chick embryos and results obtained were as follows. The rates of UV-induced unscheduled DNA synthesis were increased as increase of UV dose. And the rates were gradually decreased as the increase of time after culture, but at higher doses the decreasing tendency was remarkable. The patterns of DNA replication were changed drastically as a function of time so that in the seven day cultures the rate of $^3$H-thymidine incorporation was found to be 0.2% of the original activity. The pattern of inhibition of DNA replication by UV damage demonstrated that in cells of earlier stages there were no remarkable changes, but in cells of later stages there was significant fluctuation. Photoreactivation and the excision of pyrimidine dimer in the one day cultures showed that photoreactivation occurred immediately after UV-irradiation, but excision of pyrimidine dimer was gradually and slowly occurred. These results indicate that the differentiation of embryonic muscle cells accompanies the gradual reduction of DNA replication and unscheduled DNA synthesis, and that the photoreactivation is rapid process compared to excision repair.

• The Journal of Korean Medicine
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• v.23 no.1
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• pp.24-36
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• 2002

Objectives: The chemotherapeutic potential of Gagamgilgyung-tang for the treatment of human lung cancer, the antitumorigenic effects of Gagamgilgyung-tang on the proliferation and apoptosis of human lung cancer cell line A427 were investigated using molecular biological approaches, Methods: To determine Gagamgilgyung-tang concentrations which do not evoke cytotoxic damage to the cell line, cell viability was examined by MTT assay. To prove Gagamgilgyung-tang's antitumorigenic potential to human lung cancer, [3H]thymidine incorporation assay, trypan blue exclusion and Cpp32 protease activity assays and quantitative RT-PCR analysis were examined. Results: While A427 cells treated with $0.1-2.0{\mu\textrm{g}}/ml$ of Gagamgilgyung-tang showed no recognizable effect, marked reductions of cell viability were detected at concentrations over $5.0{\;}\mu\textrm{g}/ml$. DNA replication of A427 cells was inhibited by Gagamgilgyung-tang in a dose-dependent manner and Gagamgilgyung-tang induced the G1 cell cycle arrest through inhibition of DNA replication. Gagamgilgyung-tang triggered apoptotic cell death of A427 and enhanced the apoptotic sensitivity of the cells that were injured by a DNA damage-inducing chemotherapeutic drug etoposide. Gagamgilgyung-tang induces expression of growth-inhibiting genes such as p53 and p21/Wafl whereas it inhibited expression of growth-promoting genes such as c-Myc and Cyclin D1. Expression of a representative apoptosis-inducing gene Bax was also found to be induced by Gagamgilgyung-tang while apoptosis-suppressing Bcl-2 expression was not changed. Conclusions: Gagamgilgyung-tang could suppress the abnormal growth of tumor cells by suppressing the survival of genetically altered cells via induction of apoptosis. This study suggests that Gagamgilgyung-tang might have an antitumorigenic potential to human lung cancer cells, which might be associated with its growth-inhibiting and apoptosis-inducing properties.

• The Korean Journal of Physiology and Pharmacology
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• v.19 no.5
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• pp.421-426
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• 2015

The increased potential for vascular smooth muscle cell (VSMC) growth is a key abnormality in the development of atherosclerosis and post-angioplasty restenosis. Abnormally high activity of platelet-derived growth factor (PDGF) is believed to play a central role in the etiology of these pathophysiological situations. Here, we investigated the anti-proliferative effects and possible mechanism(s) of murrayafoline A, a carbazole alkaloid isolated from Glycosmis stenocarpa Guillamin (Rutaceae), on PDGF-BB-stimulated VSMCs. Murrayafoline A inhibited the PDGF-BB-stimulated proliferation of VSMCs in a concentration-dependent manner, as measured using a non-radioactive colorimetric WST-1 assay and direct cell counting. Furthermore, murrayafoline A suppressed the PDGF-BB-stimulated progression through $G_0/G_1$ to S phase of the cell cycle, as measured by [$^3H$]-thymidine incorporation assay and cell cycle progression analysis. This anti-proliferative action of murrayafoline A, arresting cell cycle progression at $G_0/G_1$ phase in PDGF-BB-stimulated VSMCs, was mediated via down-regulation of the expression of cyclin D1, cyclin E, cyclin-dependent kinase (CDK)2, CDK4, and proliferating cell nuclear antigen (PCNA), and the phosphorylation of retinoblastoma protein (pRb). These results indicate that murrayafoline A may be useful in preventing the progression of vascular complications such as restenosis after percutaneous transluminal coronary angioplasty and atherosclerosis.

• Journal of Periodontal and Implant Science
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• v.30 no.1
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• pp.65-77
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• 2000

The purpose of this study was to evaluate the antimicrobial activity of Crassirhzimae rhizoma and its possible use as an oral antiseptics for prevention of periodontitis. Its antibacterial activity against periodontopathic microorganisms including Actinobacillus actiomycetem - comitans, Capnocytophaga ochracea, Streptococcus mutans, Porphyromonas gingivalis, Prevotella intermedia, Actinomyces viscosus, Fusobacterium nucleatumwas evaluated via modified stab culture method. The cytotoxicity against gingival fibroblasts and rat osteoblasts was investigated via [$^3H$]thymidine incorporation and cellular activity was investigated via MTT assay. Chlorhexidine was used as control group. Crassirhizomae rhizoma was prepared at concentrations of 0.2, 0.15, 0.1, 0.05%. Chlorhexidine was also prepared at the same concentration. Crassirhizomae rhizoma showed lower antimicrobial antivity against these microorganism than chlorhexidine, but this difference was not significant. And, Crassirhzomae rhizoma showed more cellular activity and less cytotoxicity than chlorhexidine on human gingival fibrablast and rat osteoblast. This study suggests that Crassirhzomae rhizoma might be a candidate for a safe oral antiseptic for the prevention and treatment of periodontal disease.

• Journal of Life Science
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• v.22 no.12
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• pp.1595-1599
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• 2012

Cell cycle activation and progression in vascular proliferative disease represent potent therapeutic targets. Luteolin, which occurs as glycosylated forms in celery, green pepper, perilla leaf, and camomile tea, has demonstrated antimutagenic, antitumorigenic, antioxidant, and antiinflammatory properties. In this study, we investigated the effect of luteolin on the proliferation of primary cultured rat aortic vascular smooth muscle cells induced by 5% fetal bovine serum. Luteolin at concentrations of 5, 20, and $50{\mu}M$ significantly inhibited this proliferation by 29.6, 50.8, and 83.1%, respectively. The incorporation of $[^3H]$-thymidine into DNA was also inhibited by 25.8, 57.6, and 81.0%, respectively. Flow cytometry analysis of DNA content revealed that FBS-inducible cell cycle progression was blocked by luteolin. Luteolin showed no cytotoxicity in VSMCs in this experimental condition according to WST-1 assays. Luteolin may represent a potential anti-proliferative agent for treatment of angioplasty restenosis and atherosclerosis.

• YAKHAK HOEJI
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• v.35 no.3
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• pp.174-181
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• 1991

These experiments were conducted to investigate the effects of glycyrrhizin(GL) and glycyrrhetinic acid(GA) on histamine synthesis, lymphocyte blastogenesis in C57BL/6J mice splenocytes, IL-1 production, $Ca^{2+}$ uptake by macrophage-like P388D$_{1}$ cells and plaque forming cell assay against SRBC. Histamine contents, lymphocyte blastogenesis, IL-1 activity, $Ca^{2+}$ uptake and plaque forming cell were determined by enzyme isotope method, [sup 3/H]-thymidine incorporation, C3H/HeJ mouse thymocytes proliferation, the addition of 5 $\mu$Ci/ml $^{45}$Ca$^{2+}$ to P388D$_{1}$, cell suspension and assay to sheep red blood cell, respectively. Cytotoxicity, which was expressed as 50% mortality, was occurred by the addition of GL(10$^{-3}$M) and GA(10$^{-4}$M). Histamine production in mouse spleen cell culture was significantly increased by the addition of 0.25 $\mu\textrm{g}$/ml of Con A, after 48 hour incubation. Con A dependent T-lymphocyte proliferation was also enhanced by the addition of 0.25 .mu.g/ml of Con A. The effects of GL on histamine contents and T-lymphocyte proliferation were significantly decreased at high dose (10$^{-5}$M), while IL-1 activity was remarkably suppressed by 10$^{-8}$~10$^{-4}$M of GL. $Ca^{2+}$ uptake was not changed, but antibody production was increased by GL(10 mg/kg). GA inhibited histamine contents at 10$^{-9}$~10$^{-7}$ and depressed Con A (0.25 $\mu\textrm{g}$/ml) dependent T-lymphocyte proliferation at 10$^{-7}$~10$^{-5}$M of GA, but increased suboptimal dose (Con A 0.1 $\mu\textrm{g}$/ml) at 10$^{-9}$~10$^{-7}$M of GA. IL-1 activity was suppressed by 10$^{-8}$~10$^{-4}$M of GA and $Ca^{2+}$ uptake was enhanced by 10$^{-9}$~10$^{-6}$ of GA, but antibody production was not changed by GA. From the above results, it is suggested that GL and GA have immuno-regulatory action. GL decreased cell-mediated immune response, and increased humoral immune response at high dose. On the other hand, low dose of GA enhanced cell-mediated immune response, while high doses of GA decreased humoral immune reaction.