• Title/Summary/Keyword: In vivo와 In vitro

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Apoptotic Effect of Co-Treatment with Valproic Acid and HS-1200 on Human Osteosarcoma Cells (Valproic acid와 HS-1200의 병용처리가 사람골육종세포에 미치는 세포자멸사 효과에 대한 연구)

  • Kim, Duck-Han;Lee, Kee-Hyun;Kim, In-Ryoung;Kwak, Hyun-Ho;Park, Bong-Soo;Jeong, Sung-Hee;Ko, Myung-Yun;Ahn, Yong-Woo
    • Journal of Oral Medicine and Pain
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    • v.35 no.3
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    • pp.165-175
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    • 2010
  • Valproic acid (VPA) is a well-known anticonvulsive agent and has been used in the treatment of epilepsy for almost 30 years. VPA emerged in 1997 as an antineoplastic agent as well, when findings indicated the substance inhibited proliferation and induced differentiation of primitive neuroectocdermal tumor cells in vivo (Cinatl et al., 1997). Antitmor activity of VPA is associated with its targeting histone deacetylases. Bile acids and their synthetic derivatives induced apoptosis in various kinds of cancer cells and anticancer effects. It has been reported that the synthetic chenodeoxycholic acid (CDCA) derivatives showed apoptosis-inducing activity on various cancer cells in vitro. This study was undertaken to investigate the synergistic apoptotic effect of co-treatment with the histone deacetylases inhibitor, VPA and a CDCA derivative, HS-1200 on human osteosarcoma (HOS) cells. Cell viability was evaluated by trypan-blue exclusion. Induction and augmentation of apoptosis were confirmed by Hoechst staining, flow cytometry (DNA hypoploidy and MMP change), Westen blot analysis and immunofluorescent staining. In this study, HOS cells co-treated with VPA and HS-1200 showed several lines of apoptotic manifestation such as nuclear condensations, the reduction of MMP, the decrease of DNA content, the release of cytochrome c into cytosol, the translocation of AIF onto nuclei, and activation of caspase-7, caspase-3 and PARP whereas each single treated HOS cells did not. Although the single treatment of 1 mM VPA or $25\;{\mu}M$ HS-1200 for 48 h did not induce apoptosis, the co-treatment of them induced prominently apoptosis. Therefore our data provide the possibility that combination therapy of VPA and HS-1200 could be considered as a novel therapeutic strategy for human osteosarcoma.

Effects of Jeju Citrus unshiu Peel Extracts Before and After Bioconversion with Cytolase on Anti-Inflammatory Activity in RAW264.7 Cells (면역세포에서 Bioconversion 전후 제주 감귤 과피 추출물의 항염증 효과)

  • Seo, Jieun;Lim, Heejin;Chang, Yun-Hee;Park, Hye-Ryeon;Han, Bok-Kyung;Jeong, Jung-Ky;Choi, Kyoung-Sook;Park, Su-Beom;Choi, Hyuk-Joon;Hwang, Jinah
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.44 no.3
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    • pp.331-337
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    • 2015
  • Citrus and its peels, which are by-products from juice and/or jam processing, have long been used in Asian folk medicine. Citrus peels show an abundant variety of flavanones, and these flavanones have glycone and aglycone forms. Aglycones are more potent than glycones with a variety of physiological functions since aglycone absorption is more efficient than glycones. Bioconversion with cytolase converted narirutin and naringin into naringenin and hesperidin into hesperetin. Therefore, this study aimed to investigate the anti-oxidant and anti-inflammatory effects of bioconversion of Citrus unshiu (CU) peel extracts with cytolase (CU-C) in RAW264.7 cells. HPLC chromatograms showed that CU and CU-C had 23.42% and 29.39% total flavonoids, respectively. There was substantial bioconversion of narirutin to naringenin and of hesperidin to hesperetin. All citrus peel extracts showed DPPH scavenging activities in a dose-dependent manner, and CU-C was more potent than intact CU. RAW264.7 cells were pre-treated with $0{\sim}500{\mu}g/mL$ of citrus peel extracts for 4 h and then stimulated by $1{\mu}g/mL$ of lipopolysaccharide (LPS) for 8 h. All citrus peel extracts showed decreased mRNA levels and protein expression of LPS-induced inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) in a dose-dependent manner. Especially, CU-C markedly inhibited mRNA and protein expression of iNOS and COX-2 compared to intact citrus peel extracts. All citrus peel extracts showed decreased NO production by iNOS activity. This result suggests that bioconversion of citrus peel extracts with cytolase may provide potent functional food materials for prevention of chronic diseases attributable to oxidation and inflammation by boosting the anti-inflammatory effects of citrus peels.

Effects of Cultivated Wild Panax ginseng Extract on the Proliferation, Differentiation and Mineralization of Osteoblastic MC3T3-E1 Cells (산양삼(cultivated wild Panax ginseng) 추출물이 조골세포 활성에 미치는 영향)

  • Seo, Hyun-Ju;Eo, Hyun Ji;Kim, Hyun Jun;Jeon, Kwon Seok;Park, Gwang Hun;Hong, Se Chul;Jeong, Jin Boo
    • Korean Journal of Plant Resources
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    • v.33 no.4
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    • pp.227-236
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    • 2020
  • Panax ginseng C.A. Meyer (P. ginseng) is known to exert a wide range of pharmacological effects both in vitro and in vivo. Although studies on ginsenoside, antioxidant activity, and anticancer effect of the cultivated wild Panax ginseng (CWP) have been conducted, there is little research on the effect of CWP extract on bone metabolism. In this study, we investigated the potential anti-osteoporotic properties of CWP on the growth and differentiation of MC3T3-E1 cells. CWP significantly increased the viability and proliferation of MC3T3-E1 cells. CWP activated intracellular alkaline phosphatase (ALP) activity in MC3T3-E1 cells. In addition, CWP increased the mineralized nodules in MC3T3-E1 cells. Furthermore, CWP increased the expression of genes such as Runx2, ALP, OPN and OCN associated with osteoblast growth and differentiation in a dose-dependent manner.

Comparison of Direct-labeling Method of Antibody with $^{99m}Tc$ and $^{188}Re$ (농양이식백서에서 $^{99m}Tc,\;^{188}Re$ 직접표지항체의 비교)

  • Choi, Tae-Hyun;Lim, Sang-Moo;Choi, Chang-Woon;Woo, Kwang-Sun;Chung, Wee-Sup;Lim, Soo-Jeong
    • The Korean Journal of Nuclear Medicine
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    • v.33 no.1
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    • pp.84-93
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    • 1999
  • Purpose: We investigated the direct labeling method of antibody with $^{99m}Tc$ and $^{188}Re$ and examined the stability and function of these labeled compounds in in vitro and in vivo. Materials and Methods: Disulfide bond of nonspecific human IgG was reduced to -SH group by 2-mercaptoethanol. Stannous ion was used to reduce $^{99m}Tc$ and $^{188}Re$. The stability of $^{99m}Tc$-IgG and $^{188}Re$-IgG was estimated upto 24 hrs. Biodistribution was evaluated in abscess bearing rats at 4 and 24 hr post-injection of $^{99m}Tc$ or $^{188}Re$ labeled IgG. Results: The number of -SH group per reduced IgG molecule was 2.34. The labeling yield of $^{99m}Tc$-IgG and $^{188}Re$-IgG were 90% and 95%, respectively The stability of $^{99m}Tc$-IgG at 1, 4, 6 and 24 hr was 91%, 83%, 78%, 7% and that of $^{188}Re$-IgG at 1, 4, 16 and 24 hr was 94%, 80%, 47%, 42%, respectively. At 4 hr post-injection of $^{99m}Tc$-IgG, high uptake was found on kidney, blood, stomach and abscess ($9.42{\pm}0.68,\;1.43{\pm}0.24,\;0.86{\pm}0.18,\;0.72{\pm}0.10$ %ID/g, respectively). The uptakes at 24 hr were kidney, abscess,.itomach, and blood in descending order. In case of $^{188}Re$-lgG, high uptake at 4 hr post injection appeared on kidney, blood, abscess and stomach ($3.92{\pm}0.62,\;1.32{\pm}0.08,\;0.88{\pm}0.01,\;0.26{\pm}0.06$, respectively). The uptakes at 24 hr were kidney, abscess, blood and stomach in descending order. The abscess to blood uptake ratio of $^{99m}Tc$-IgG was 0.5 at 4 hr and 2.02 at 24 hr and that of $^{188}Re$-IgG was 0.67 and 1.29. Conclusion: $^{99m}Tc$-IgG and $^{188}Re$-IgG canbe labeled efficiently with direct labeling method. However, $^{99m}Tc$-IgG and $^{188}Re$-IgG, labeled with direct method, was unstable. Further study is needed to enhance the stability of the antibody labeling.

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Screening for Effective Organic Farming Materials for the Control of Cucumber Scab Caused by Cladosporium cucumerinum (오이 검은별무늬병 방제에 효과적인 유기농업자재 선발)

  • Park, So-Hyang;Hong, Sung-Jun;Shim, Chang-Ki;Kim, Min-Jeong;Park, Jong-Ho;Han, Eun-Jung;Kim, Yong-Ki
    • Research in Plant Disease
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    • v.23 no.2
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    • pp.159-167
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    • 2017
  • This study was conducted to evaluate the control efficacy of the organic farming materials on cucumber scab caused by Cladosporium cucumerinum PT1 (KACC 48094). The antifungal activities in vitro as well as the suppressive effect of 43 organic farming materials on the spore germination and germ tube growth by inoculating spore suspension on cucumber seedlings in vivo were investigated. Thirteen organic farming materials inhibited the mycelial growth of C. cucumerinum and nine of these were microbial agents. In the screening using cucumber seedlings, six organic farming materials were very effective with control efficacy value of 90%. Among them, Bacillus amyloliquefaciens M27 provided suppressive effect on both mycelial growth and spore germination against cucumber scab. Finally, nine organic farming materials were selected to test the protective and curative effects, and all chosen organic farming materials significantly suppressed disease incidence when applied in the preventive action, in comparison with the curative action. Especially, Bordeaux mixture I and III gave excellent protective control efficacy with control values of 96.7% and 73.3%, respectively, whereas its curative control effect was significant low. Among these, only Thymus quinquecostatus+Sophora extract showed curative activity, although the control value was as low as 50%. This study suggests that cucumber scab can be controlled by some organic farming materials in the farmhouses under comparatively cold and wet condition and protective treatment is more important and efficient.

The effect of two Terpenoids, Ursolic acid and Oleanolic acid on epidermal permeability barrier and simultaneously on dermal functions

  • Lim Suk Won;Jung Sung Won;Ahn Sung Ku;Kim Bora;Ryoo Hee Chang;Lee Seung Hun
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.29 no.2 s.43
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    • pp.205-232
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    • 2003
  • Ursolic acid (UA) and Oleanolic acid (ONA), known as urson, micromerol and malol, are pentacyclic triterpenoid compounds which naturally occur in a large number of vegetarian foods, medicinal herbs, and plants. They may occur in their free acid form or as aglycones for triterpenoid saponins, which are comprised of a triterpenoid aglycone, linked to one or more sugar moieties. Therefore UA and ONA are similar in pharmacological activity. Lately scientific research, which led to the identification of UA and ONA, revealed that several pharmacological effects, such as antitumor, hepato-protective, anti-inflammatory, anticarcinogenic, antimicrobial, and anti-hyperlipidemic could be attributed to UA and ONA. Here, we introduced the effect of UA and ONA on acutely barrier disrupted and normal hairless mouse skin. To evaluate the effects of UA and ONA on epidermal permeability barrier recovery, both flanks of 8-12 week-old hairless mice were topically treated with either 0.01-0.1 mg/ml UA or 0.1-1 mg/ml ONA after tape stripping, and TEWL (Transepidermal water loss) was measured . The recovery rate increased in those UA or ONA treated groups (0.1 mg/ml UA and 0.5 mg/ml ONA) at 6 h more than $20\%$ compared to vehicle treated group (p<0.05). Here, we introduced the effects of UA and ONA on acute barrier disruption and normal epidermal permeability barrier function. For verifying the effects of UA and ONA on normal epidermal barrier, hydration and TEWL were measured for 1 and 3 weeks after UA and ONA applications (2mg/ml per day). We also investigated the features of epidermis and dermis using electron microscopy (EM) and light microscopy (LM). Both samples increased hydration compared to vehicle group from f week without TEWL alteration (p<0.005). EM examination using RuO4 and OsO4 fixation revealed that secretion and numbers of lamellar bodies and complete formation of lipid bilayers were most prominent $(ONA{\geq}UA>Vehicle)$. LM finding showed that thickness of stratum corneum (SC) was slightly increased and especially epidermal thickening and flattening was observed (UA>ONA>Veh). We also observed that UA and ONA stimulate epidermal keratinocyte differentiation via $PPAR\;\alpha$. Protein expression of involucrin, loricrin, and filaggrin increased at least 2 and 3 fold in HaCaT cells treated with either $ONA\;(10{\mu}M)$ or UA $(10{\mu}M)$ for 24h respectively. This result suggested that the UA and ONA can improve epidermal permeability barrier function and induce the epidermal keratinocyte differentiation via $PPAR\;{\alpha}$. Using Masson-trichrome and elastic fiber staining, we observed collagen thickening and elastic fiber elongation by UA and ONA treatments. In vitro results of collagen and elastin synthesis and elastase inhibitory activity measurements were also confirmed in vivo findings. These data suggested that the effects of UA and ONA related to not only epidermal permeability barrier functions but also dermal collagen and elastic fiber synthesis. Taken together, UA and ONA can be relevant candidates to improve epidermal and dermal functions and pertinent agents for cosmeseutical applications.

Anti-inflammatory effects of Ishige sinicola ethanol extract in LPS-induced RAW 264.7 cell and mouse model (LPS로 유도된 RAW 264.7 Cell과 마우스 모델에 대한 넓패(Ishige sinicola) 에탄올 추출물의 항염증 효과)

  • Kim, Ji-Hye;Kim, Min-Ji;Kim, Koth-Bong-Woo-Ri;Park, Sun-Hee;Cho, Kwang-Su;Kim, Go-Eun;XU, Xiaotong;Lee, Da-Hye;Park, Ga-Ryeong;Ahn, Dong-Hyun
    • Food Science and Preservation
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    • v.24 no.8
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    • pp.1149-1157
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    • 2017
  • Inflammation is the first response of the immune system to infection or irritation in our body. The use of medicinal plants has been widely applied as an alternative source for drug development. One of marine natural resources, the anti-inflammatory effect of Ishige sinicola ethanol extract (ISEE), was evaluated by using LPS-induced RAW 264.7 cell and mice model. As a result, the production of nitric oxide (NO) and pro-inflammatory cytokines (IL-6, IL-$1{\beta}$, TNF-${\alpha}$) were inhibited with increasing concentration of ISEE without any cytotoxicity. Furthermore, ISEE suppressed the expression of not only inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), nuclear factor-kappa B (NF-${\kappa}B$) p65, and mitogen-activated protein kinases (MAPKs), including extracellular signal-regulated kinase (ERK) 1/2, p38, and c-Jun N-terminal kinase (JNK) in a dose-dependent manner. In mice ear edema test, the formation of edema was reduced at the highest dosage of ISEE and the reduction of the number of infiltrated mast cells was observed in histological analysis. These results indicate that ISEE has a potent anti-inflammatory activity and can be used as a pharmaceutical material for many kinds of inflammatory disease.

Histological studies on in vitro Propagation of Pulsatilla koreana Nakai (할미꽃 기내증식(器內增殖)에 관(關)한 조직학적(組織學的) 연구(硏究))

  • Lee, Man-Sang;Oh, Ki-Hong
    • Korean Journal of Medicinal Crop Science
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    • v.1 no.2
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    • pp.137-157
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    • 1993
  • This study was carried out to investigate the optimal condition for multiple propagation through leaf tissue culture and to apply anther culture techniques to Pulsatilla koreana Nakai breeding. Leaf and anther of Pulsatilla koreana Nakai were cultured on MS, MT, LS and $B_5$ media supplemented with several growth regulators and nitrogen sources under various conditions. For callus induction and differentiation from the Pulsatilla koreana leaf segments were more effective in the combination of zeatin and auxin than auxin alone. The color of the callus was green when treated with IBA alone. Shoot differentiation was more effective when treated with zeatin than auxin alone, especially the best hormoal combination for shoot differentiation was zeatin 1.0mg/l +NAA 0.1mg/l, while 2,4-D inhibited shoot differentiation. The appeared rate of S pollen was 35% in vivo, while that of S pollen by low temperature$(4^{\circ}C)$ pretreatment for 4 days was increased by 53% and the optimum culture time for callus induction from anther was uni-nucleate stage. $B_5$ basal medium supplemented with NAA 0.5mg/l and zeatin 1 mg/l was the most effective on callus formation and the best results of plant regeneration were obtained from combination of NAA 0.5mg/l and zeatin 0.5mg/l in anther culture. $NH_{4}NO_3$ as more effectives as the nitrogen source than $KNO_3$ and the combination with zeatin 2.0mg /L was the best effective. The best combination for plant regeneration in callus induced from anther was $NH_{4}NO_3$ 1650mg/l + $KNO_3$ 3800mg/l + zeatin 2.0mg/l. Ploidy level of anther-derived plants appeared 28% haploid, 47% diploid and the others were triploid, tetraploid and mixploid. In compare with E.S.T, M.D.H and P.X banding patterns were distinguished among callus, haploid and diploid plants in electrophoresis.

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Effect of Steroid Administration Ex Vivo on the IκB/NF-κB Pathway in Human Peripheral Blood Monocytes (스테로이드의 투여가 말초혈액 단핵구에서 IkB/NF-κB경로에 미치는 영향)

  • Yoon, Ho Il;Lee, Hee-Seok;Lee, Chang-Hoon;Lee, Choon-Taek;Kim, Young Whan;Han, Sung Koo;Shim, Young-Soo;Yoo, Chul-Gyu
    • Tuberculosis and Respiratory Diseases
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    • v.54 no.5
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    • pp.542-550
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    • 2003
  • Background : Synthetic glucocorticoids are widely used in many chronic inflammatory diseases because of their excellent anti-inflammatory activity. Enhancing the transcription of $I{\kappa}B$ and preventing activated NF-${\kappa}B$ from binding to ${\kappa}B$ sites are thought to be the underlying mechanisms. But these data are largely derived from in vitro studies using cell lines. In this study, after administrating a steroid to volunteers, we evaluated the effect on the NF-${\kappa}B$ system. Methods : Prednisolone(0.5mg/kg/d) was orally administered to 5 healthy volunteers for 7 days. Before and after the administration, we sampled their peripheral blood monocytes, and performed western blot analysis both with stimulation, using IL-$1{\beta}$, LPS, TNF, and without stimulation(baseline). We also performed EMSA after stimulation with LPS. Results : After ingestion of the steroid, baseline expressions of $I{\kappa}B{\alpha}$ were increased in two of the subjects, while suppressed degradations of $I{\kappa}B{\alpha}$ to stimulations were observed in all five. In addition, the binding capacity of NF-${\kappa}B$ after the administration was decreased. Conclusion : Steroid plays such roles as enhancing the transcription of $I{\kappa}B{\alpha}$, suppressing the DNA binding capacity of NF-${\kappa}B$, and suppressing the degradation of $I{\kappa}B{\alpha}$.

Anti-Stress Effect of Punica granatum L. Extract against Sleep Deprivation-Induced Impairment (석류 열수 추출물의 수면박탈을 유도한 Rat 모델에서의 항스트레스 효과)

  • Na, Ju-Ryun;Kim, Sunoh;Jo, Ara;Bae, Donghyuck;Oh, Kyo-Nyeo;Kim, Yong Jae;Lee, Yoo-Hyun;Jun, Woojin
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.45 no.11
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    • pp.1533-1543
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    • 2016
  • The anti-stress effects of Punica granatum L. (family Lythraceae, PG) on $H_2O_2$/corticosterone (CORT)-induced stress in cells and sleep-deprived rats were investigated. The PG extract showed neuroprotective effects in SH-SY5Y cells against $H_2O_2$/CORT-induced stress. Sleep deprivation led to behavioral, hormonal, and biochemical alterations in the animal model. The effects of P. granatum on physiological, behavioral, and biochemical parameters aggravated by sleep deprivation were investigated. Sleep deprivation impaired physiological (survival, body weight, and drowsiness scores) and behavioral (rotarod, passive avoidance, hot hyperalgesia, and Y maze) parameters as well as biochemical factors (cortisol, serotonin, dopamine, testosterone, and growth factor I contents in serum). These parameters were significantly recovered by PG extract in a concentration-dependent manner. The PG extract also enhanced catalase, superoxide dismutase, and non-enzymatic antioxidative activities such as glutathione compared to sleep-deprived rats. On the basis of these results, our findings suggest that Punica granatum prevents impairment of body functions induced by sleep deprivation and related oxidative damage.