• 한방안이비인후피부과학회지
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• 제12권1호
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• pp.47-78
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• 1999

Hoichunyonggyuksan(HCYGS) and Yongseksan(YSS) are important prescriptions that have been used in oriental medicine for stomatitis and wound healing. The study was done to evaluate the inhibitory effects of cytotoxicity, formation of superoxide on the macrophage and neutrophil, prostaglandins($PGE_2$), interleukins($IL-1{\Beta}$), collagenase activity and synthesis of collagen and DNA. The results were obtained as fo1lows: 1. HCYGS and YSS were not showed the proliferation difference of human fibroblast and monocyte in all concentrations to be experimented and in result, it was concluded that they have no cytotoxicity. 2. YSS inhibited the formation of superoxide to $48\% at the concentration of $0.001\%$ in the mouse monocyte. 3. HCYGS inhibited the formation of superoxide to $13\%$ at the concentration of $0.001\%$ as compared with control in the human monocyte. 4. HCYGS and YSS inhibited the formation of superoxide to $25\%\;and\;35\%$ respectively at the concentration of $0.0001\%\;and\;HCYGS\;showed\;38\%$ inhihitory rate on the formation of superoxide at concentration of $0.001\%$ in the human neutrophil. 5. The concentration of inhibiting the production of prostaglandins($PGE_2$) to $11\%$ in the human monocyte stimulated with E. coli were $0.01\%$ of HCYGS. 6. The concentration of inhibiting the production of interleukins($IL-l{\Beta}$) to $43\%\;and\;39\%$ in the human monocyte stimulated with E. coli were $0.01\%$ of HCYGS and YSS. 7. HCYGS and YSS didn't influence on collagen synthesis and total protein in fibroblasts. 8. HCYGS and YSS inhibited the collagenase activity to $22\%\;and\;19\%\;at\;0.1\%,\;45\%\;and\;56\%\;at\;0.2\%,\;57\%\;and\;52\%$ at $0.5\%$ respectively, but YSS exerted the inhibitory effect on collagenase activity to $27\%$ at the lower concentration of $0.01\%$. 9. HCYGS and YSS didn't show the faster recovary than control group hut exerted the consistant effect on the anti-inflammations and the formation of granulation tissue.

• IMMUNE NETWORK
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• 제7권1호
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• pp.10-17
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• 2007

Autoimmune arthritis, such as rheumatoid arthritis (RA), is a chronic inflammatory disorder that primarily affects the joints and then results in their progressive destruction. Effector Th cells have been classified as Th1 and Th2 subsets based on their cytokine expression profiles and immune regulatory function. Another subset of T cells termed Th17 was recendy discovered and known to selectively produce IL-17. Also, Th17 was shown to be generated by TGF${\beta}$ and IL-6 and maintained by IL-23. IL-17 is a proinflammatory cytokine that is considered to involve the development of various inflammatory autoimmune diseases such as RA, asthma, lupus, and allograft rejection. IL-17 is present in the sera, synovial fluids and synovial biopsies of most RA patient. IL-17 activates RA synovial fibroblasts to synthesize IL-6, IL-8 and VEGF via PI3K/Akt and NF-${\kappa}B$ dependent pathway. IL-17 increases IL-6 production, collagen destruction and collagen synthesis. In addition, it not only causes bone resorption but also increases osteoclastogenesis and fetal cartilage destruction. Inhibition of the IL-17 production may contribute a novel therapeutic approach along with potent anti-inflammatory effect and with less immunosuppressive effect on host defenses.

• Asian Pacific Journal of Cancer Prevention
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• 제15권23호
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• pp.10397-10400
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• 2015

Stigmalactam, an aristolactam-type alkaloid extracted from Orophea enterocarpa, exerts cytotoxicity against several human and murine cancer cell lines, but the molecular mechanisms remain elusive. The aims of this study were to identify the mode and mechanisms of human cancer cell death induced by stigmalactam employing human hepatocellular carcinoma HepG2 and human invasive breast cancer MDA-MB-231 cells as models, compared to normal murine fibroblasts. It was found that stigmalactam was toxic to HepG2 and MDA-MB-231 cells with $IC_{50}$ levels of $23.0{\pm}2.67{\mu}M$ and $33.2{\pm}4.54{\mu}M$, respectively, using MTT assays. At the same time the $IC_{50}$ level towards murine normal fibroblast NIH3T3 cells was $24.4{\pm}6.75{\mu}M$. Reactive oxygen species (ROS) production was reduced in stigmalactam-treated cells dose dependently after 4 h of incubation, indicating antioxidant activity, measured by using 2',7',-dichlorohydrofluorescein diacetate and flow cytometry. Caspase-3 and caspase-9 activities were increased in a dose response manner, while stigmalactam decreased the mitochondrial transmembrane potential dose-dependently in HepG2 cells, using 3,3'-dihexyloxacarbocyanine iodide and flow cytometry, indicating mitochondrial pathway-mediated apoptosis. In conclusion, stigmalactam from O. enterocarpa was toxic to both HepG2 and MDA-MB-231 cells and induced human cancer HepG2 cells to undergo apoptosis via the intrinsic (mitochondrial) pathway.

• 한국식품과학회지
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• 제38권1호
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• pp.114-120
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• 2006

백련 뿌리 및 잎과 조릿대는 과거부터 약용으로 사용해 왔지만 아직까지 연구도 많이 이루어지지 않았고, 항당뇨 효과에 대한 연구도 거의 없었다. 본 연구에서는 백련 뿌리 및 잎과 조릿대의 추출물과 분획물이 in vitro에서 인슐린 작용, 인슐린 분비 또는 탄수화물의 소화에 효과적인지를 조사함으로 항당뇨에 효과적인지 여부를 조사하였다. 백련 뿌리 및 잎과 조릿대의 추출물은 각각 물로 추출하여 항당뇨 효과를 조사하였다. 또한 백련 뿌리 및 잎과 조릿대의 3 : 2 : 3으로 혼합하여 물로 추출한 후 이를 메탄올과 물을 섞은 용액으로 단계별로 XAD-4 column으로 분획하였다. 백련 뿌리 및 잎과 조릿대의 추출물과 혼합물의 분획물은 고농도(1 mg/mL)에서도 MTT 방법으로 측정하였을 때 세포 독성을 나타내지 않았다. 백련 뿌리와 조릿대 물추출물은 인슐린 작용을 향상시키는 효능이 있었고, 백련잎 물추출물은 ${\alpha}-amylase$를 억제하여 탄수화물의 소화 흡수를 지연시켰다. 이에 백련 뿌리 및 잎과 조릿대를 3 : 2 : 3으로 혼합하였을 때 20과 80% 메탄올층은 3T3-L1 지방세포에 처리하였을 때 인슐린의 작용을 향상시켜 포도당의 흡수를 증가시키는 효과가 인슐린을 10 nM을 처리한 것 만큼 효과적으로 포도당 흡수를 증가시켰다. 이 층에는 3T3-L1 섬유아세포에 분화 유도물질과 함께 처리하였을 때 $PPAR-{\gamma}$ agonist인 rosiglitazone과 마찬가지로 지방 세포로의 분화를 촉진시키고 지방의 축적도 증가시켰다. 그러므로 80% 메탄올 층에는 $PPAR-{\gamma}$ agonist로 작용하는 물질이 함유되어 있을 가능성이 높다. 베타세포라인인 Min6 세포에 백련 뿌리 및 잎과 조릿대의 혼합물의 분획물을 처리한 후 저농도와 고농도 포도당 자극시 인슐린 분비를 측정하였을 때 두 농도에서 모두 인슐린 분비에 영향을 미치지 않았다. 또한 백련 뿌리 및 잎과 조릿대의 혼합물의 20, 60과 80% 메탄올 분획층은 탄수화물의 소화에 작용하는 효소인 ${\alpha}-amylase$의 활성을 16% 정도를 억제하는 효과가 있었다. 결론적으로 백련 뿌리 및 잎과 조릿대의 추출물과 분획물에는 인슐린 분비나 탄수화물의 소화에 관여하는 성분이 없지만, 인슐린 작용을 향상시키는 인슐린 민감성 물질이 함유되어 있을 가능성이 높다.

• Biomolecules & Therapeutics
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• 제16권2호
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• pp.87-94
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• 2008

In view of obtaining potential anticancer compounds, we studied the inhibitory activity and the cytotoxic effects of a candidate compound in cancer cells. The cytotoxic effects of cannabidiol (CBD) in vitro were evaluated in NIH3T3 fibroblasts, B16 melanoma cells, A549 lung cancer cells, MDA-MB-231 breast cancer cells, Lenca kidney cells and SNU-C4 colon cancer cells. The cells were cultured in various concentrations of CBD for 48 h and 25 ${\mu}$M of CBD for 6-36 h. The cells were observed to exhibit inhibitory effects of the cell viability in their growth, and then cytotoxicity was estimated. The inhibitory activity of CBD was increased in all cancer cells and showed especially strong increment in breast cancer cells. The cytotoxicity of CBD increased in a dose- and time-dependent manner with growth inhibition in all cancer cell lines. Also, to assess the membrane toxicity induced by CBD, we investigated lactate dehydrogenase (LDH) release. After treatment with various concentrations of CBD, LDH release rate of cancer cells was accelerated. On the other hand, in the induction of cell death, caspase-3, -8 and -9 activations were detected in cancer cells after treatment with various concentrations of CBD, and CBD effectively induced activity of caspase-3, -8 and -9 in A549 lung cancer cells, MDAMB-231 breast cancer cells and Renca kidney cells. Therefore these results suggest that CBD has a possibility of anticancer agents and anticancer effects against cancer cells by modulation of apoptotic pathway in the range of 5-80 ${\mu}$M concentration.

• 농약과학회지
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• 제6권2호
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• pp.96-104
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• 2002

Paraquat의 세포독성을 알아보기 위하여 NIH 3T3 섬유모세포에 적용한 후 MTT와 NR 분석법을 이용하여 세포독성을 측정하고, paraquat의 세포독성에 대한 3-MC의 독성경감효과를 알아보기 위하여 Spraque Dawley계 수컷 랫드에 paraquat 단독 및 paraquat와 3-MC 병용투여 후 랫드의 폐를 경시적으로 채취하여 관찰하였다. Paraquat의 NIH 3T3 섬유모세포에 대한 $MTT_{50}$은 $1668.97{\mu}M$, $NR_{50}$은 $1030.25{\mu}M$로 산출되어 Borefreund와 Puemer(1984)의 독성판정기준에 의하면 저독성 물질이었다. Paraquat 단독 투여 군은 H&E 염색에서 3시간째부터 폐 모세혈관 내에 적혈구 수가 증가하기 시작하여 24시간째에는 충혈상태에 이르렀으며, 폐포사이 중격에서는 큰폐포상피수가 증가하였다. 또한 폐 조직을 둘러싸고 있는 결합조직 내에는 임파구, 대식세포 및 다형핵 백혈구 등이 다수 관찰되었고, 48시간째부터 폐포사이 중격과 폐포내에 폐포큰포식세포가 증가하기 시작하여 96시간째에는 다수의 폐포큰포식세포가 관찰되었다. Verhoeff의 iron hematoxylin 염색에서도 paraquat 단독 투여 후 24시간째에 조직변화가 가장 심하였고, 교원섬유량의 급격한 증가, 폐포의 넓이와 폐포 구멍(alveolar pore) 간격의 확장 등이 관찰되었다. 한편, paraquat와 3-MC 병용투여군은 paraquat 단독 투여 군에 비하여 조직변화가 약하게 관찰되었는데, 병용투여 후 3시간째에는 단독투여 3시간째의 소견과 유사하였으나 점차 회복되어 폐 모세혈관 내에 적혈구 수가 증가하여 24시간째에는 대조군의 구조와 거의 유사하였다. 또한 폐 조직을 둘러싸고 있는 결합조직과 임파소절에서도 paraquat 단독 투여 군에서 보였던 변화가 거의 관찰되지 않았다. Verhoeff의 iron hematoxylin 염색에서도 병용투여 후 24시간째에는 교원섬유량이 단독 투여 군에 비하여 크게 감소하였고 폐포와 폐포 구멍의 넓이도 대조군과 유사하였다.

• 생명과학회지
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• 제28권6호
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• pp.681-687
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• 2018

홍해삼 또는 Apostichopus japonicas는 동남아시아에서 발견되는 stichopodiae의 한 종이다. 본 연구에서는 홍해삼의 화장품 소재로서 사용가능한지 알아보기 위해 홍해삼 추출물의 미백, 항주름에 관한 실험을 진행하였다. tyrosinase 활성 분석 결과, 홍해삼 추출물 $200{\mu}g/ml$에서 tyrosinase 활성을 억제하였다. 또한, 홍해삼은 tyrosinase, TRP-1, TRP-2, MITF 및 matrix metalloproteinase (MMPs)의 mRNA 발현을 억제하였다. 이어서 HaCaT과 human Fibroblast를 이용한 3차원 세포배양을 통해 홍해삼의 피부 턴오버 주기 개선효과를 검증하였다. 이러한 결과를 바탕으로 홍해삼이 높은 미백효과 및 주름개선효과를 가지는 화장품 소재로서 충분한 가치를 지닐 것으로 판단된다.

• Reproductive and Developmental Biology
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• 제28권1호
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• pp.7-12
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• 2004

This study was performed to establish the effective culture condition for the establishment of clonal lines from porcine fetal fibroblast cells. Fibroblasts derived from a pig fetus (Day 50) were cultured and passaged two times before use. A single cell was seeded in 96-well plates, cultured in medium supplemented with different concentrations of FBS, catalase or $\beta$-mercaptoethanol ($\beta$ME), and classified by cell size and morphology. Cells were passaged two times into 4-well dish before freezing. The establishment efficiencies were not different among different concentrations of FBS (0.3 to 5.1%). However, population doubling time (PDT) was significantly decreased by increasing the FBS concentration (P＜0.05). The establishment efficiency of $\beta$ME-added group (10.4%) was significantly higher than those of catalase-added and control groups (3.5%, and 3.5%, respectively, p＜0.05), and PDT was significantly decreased (23.6 vs 28.1, and 25.5 h, respectively, p＜0.05). However, catalase did not show a positive effect on the establishment efficiency. Cell size and morphology did not affect the establishment efficiency and PDT of clonal lines. The result of present study shows that the establishment efficiency of clonal cell lines can be enhanced by the culture in media supplemented with 30% FBS and $\beta$ME.

• BMB Reports
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• 제34권5호
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• pp.484-488
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• 2001

STATs are proteins with a dual function: signal transducers in the cytoplasm and transcriptional activators in the nucleus. Among the six known major STATs (STAT1-6), STAT3 has been implicated in the widest range of signaling pathways that regulate cell growth and differentiation. As a part of our on-going investigation on the pleiotropic functions of STAT proteins, we examined the role of STAT3 as a molecular adaptor that links diverse cell growth signaling pathways. We observed that STAT3 can be specifically activated by multiple cytokines, such as IL-3, in transformed fibroblasts and IL-4 or IFN-$\gamma$ in primary immune cells, respectively. The selective activation of STAT3 in H-ras-transformed NIH3T3 cells is associated with an increased expression of phosphoserioe STAT3 in these cells, compared to the parental cells. Notably phosphoresine-STAT3 interacts with oncogenic ras, shown by immunoprecipitation and Western blots. The results suggest the role of STAT3 in rasinduced cellular transformation as a molecular adaptor linking the Jak/STAT and Ras/MAPK pathways. In primary immune cells, IL-4 and IFN-$\gamma$ each induced (in addition to the characteristic STAT6 and STAT1 homodimers) the formation of STAT3-containing complexes that bind to GAS probes, which correspond to the $Fe{\varepsilon}$ Rll and $Fe{\gamma}$ RI promoter sequences, respectively. Since IL-4 and IFN-$\gamma$ are known to counter-regulate the expression of these genes, the ability of STAT3 to form heterodimeric complexes with STAT6 or STAT1 implies its role in the fine-tuned control of genes that are regulated by IL-4 and IFN-$\gamma$.

• The Journal of Advanced Prosthodontics
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• 제12권2호
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• pp.89-99
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• 2020

PURPOSE. The effects of four different self-adhesive resin cement materials on cell viability and apoptosis after direct and indirect exposure were evaluated using different cell culture techniques. MATERIALS AND METHODS. Self-adhesive cements were applied to NIH/3T3 mouse fibroblasts by the extract test method, cell culture inserts, and dentin barrier test method. After exposure periods of 24 h and 72 h, the cytotoxicity of these self-adhesive materials was evaluated using the MTT assay (viability) and the Annexin-V-FITC/PI staining (apoptosis). RESULTS. The lowest cell viability was found in cells exposed to BeautiCem SA for 24 h in the extract test method. Cell viability was reduced to 70.6% compared to negative controls. After the 72 h exposure period, viability rate of cell cultures exposed to BeautiCem SA decreased more than 2- fold (29.5%) while cells exposed to RelyX U200 showed the highest viability rate of 71.4%. In the dentin barrier test method, BeautiCem SA induced the highest number of cells in apoptosis after a 24 h exposure (4.1%). Panavia SA Cement Plus was the material that caused the lowest number of cells in apoptosis (1.5%). CONCLUSION. The used self-adhesive cements have showed different cytotoxic effects based on the evaluation method. As exposure time increased, the materials showed more cytotoxic and apoptotic effects. BeautiCem SA caused significantly more severe cytotoxic and apoptotic effects than other cements tested. Moreover, cements other than BeautiCem SA have caused necrotic cell death rather than apoptotic cell death.