• Title/Summary/Keyword: in-vitro

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Rec-assay and DNA-breaking Action on the Engymatic Browning Reaction Products -Influence of Cupric Ion Concentration- (효소적(酵素的) 갈변반응(褐變反應) 생성물(生咸物)에 대(對)한 Rec-assay 및 DNA 절단(切斷) 작용(作用) -동(銅) Ion 농도(濃度)의 영향(影響)-)

  • Ham, S.S.;Park, B.K.;Lee, S.Y.;Lee, J.H.;Kang, C.K.;Lee, D.S.;Omura, Hirohisa
    • Applied Biological Chemistry
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    • v.27 no.4
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    • pp.264-270
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    • 1984
  • In order to obtain mutagenic data of enzymatic browning reaction products, we investigated their mutagenicity. The rec-assay with Bacillus subtilis strains $H17(rec^+)$ and $M45(rec^-)$ were carried out with their spores. Detection of double strand breakage in calf-thymus DNA was investigated into sample solution with and without $Cu^{2+}$ by the agarose slab gel electrophoresis. In the rec-assay, catechol, pyrocatechol, DL-dopa, 3,4-dihydroxytoluene, hydroquinone, hydroxyhydroquinone with and without $Cu^{2+}$ in 0.05M ana 0.1M at the enzymatic browning reaction products stowed mutagenic action. And also browning solution of 0.05M hydroxyhydroquinone and catechol with $Cu^{2+}$, hydroquinone with and without $Cu^{2+}$ of 0.1M at the enzymatic browning reaction products were strong mutagenic action. The DNA breakability of the enzymatic browning reaction products of 0.1M pyrogallol was stronger than that of 0.05M pyrogallol browning solution with $Cu^{2+}$ and 3,4-dihydroxytoluene browning solution was stronger than that of 0.01M 3,4-dihydroxytoluene browning solution.

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EFFECT OF CHLORHEXIDINE ON MICROTENSILE BOND STRENGTH OF DENTIN BONDING SYSTEMS (Chlorhexidine 처리가 상아질 접착제의 미세인장결합강도에 미치는 영향)

  • Oh, Eun-Hwa;Choi, Kyoung-Kyu;Kim, Jong-Ryul;Park, Sang-Jin
    • Restorative Dentistry and Endodontics
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    • v.33 no.2
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    • pp.148-161
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    • 2008
  • The purpose of this study was to evaluate the effect of chlorhexidine (CHX) on microtensile bond strength (${\mu}TBS$) of dentin bonding systems. Dentin collagenolytic and gelatinolytic activities can be suppressed by protease inhibitors, indicating that MMPs (Matrix metalloproteinases) inhibition could be beneficial in the preservation of hybrid layers. Chlorhexidine (CHX) is known as an inhibitor of MMPs activity in vitro. The experiment was proceeded as follows: At first, flat occlusal surfaces were prepared on mid-coronal dentin of extracted third molars. GI (Glass Ionomer) group was treated with dentin conditioner, and then, applied with 2 % CHX. Both SM (Scotchbond Multipurpose) and SB (Single Bond) group were applied with CHX after acid-etched with 37% phosphoric acid. TS (Clearfil Tri-S) group was applied with CHX, and then, with adhesives. Hybrid composite Z-250 and resin-modified glass ionomer Fuji-II LC was built up on experimental dentin surfaces. Half of them were subjected to 10,000 thermocycle, while the others were tested immediately. With the resulting data, statistically two-way ANOVA was performed to assess the ${\mu}TBS$ before and after thermo cycling and the effect of CHX. All statistical tests were carried out at the 95 % level of confidence. The failure mode of the testing samples was observed under a scanning electron microscopy (SEM). Within limited results, the results of this study were as follows; 1. In all experimental groups applied with 2 % chlorhexidine, the microtensile bond strength increased, and thermo cycling decreased the micro tensile bond strength (P > 0.05). 2. Compared to the thermocycling groups without chlorhexidine, those with both thermocycling and chlorhexidine showed higher microtensile bond strength, and there was significant difference especially in GI and TS groups. 3. SEM analysis of failure mode distribution revealed the adhesive failure at hybrid layer in most of the specimen. and the shift of the failure site from bottom to top of the hybrid layer with chlorhexidine groups. 2 % chlorhexidine application after acid-etching proved to preserve the durability of the hybrid layer and microtensile bond strength of dentin bonding systems.

Utilization of Obosan (Dietary Herbs) I. Effects on Survival, Growth, Feed Conversion Ratio and Condition Factor in Olive Flounder, Paralichthys olivaceus (한방사료 첨가제인 어보산의 효과 I. 넙치의 생존율, 성장, 사료효율 및 비만도에 미치는 영향)

  • 김동수;김종현;정창화;이상윤;이상민;문영봉
    • Journal of Aquaculture
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    • v.11 no.2
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    • pp.213-221
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    • 1998
  • The effects of differen concentrations of Obosan as a feed additive dietary herb were examined on survival rate, growth, feed conversion ration and condition factor in olive flounder (paralichthys olivaceus). Effectiveness of dietary Obosan with optimized concentration for 48 weeks were also observed with regard to growth performances and yields. All groups fed diets containing 0.15, 0.3 and 0.6% of Obosan revealed significantly higher survival rate than control group (P<0.05). Growth, feed conversion ratio and condition factor of olive flounder fed diets containing Obosan were considerably improved when compared to those of controls (P<0.05). The 0.3% of dietary Obosan was proven to be the optimal concentration in all parameters tested. The dietary Obosan (0.3%) for 48 weeks showed significantly higher surval rate than control (P<0.05), and also improved yields in weight gain (19.0% improvement), specific growth rate (4.8%), feed conversion ratio(13.6%) and condition factor (10.8%), significantly (P<0.05).

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The Effect of Histamine on Polymorphonuclear Leukocyte-induced Pneumocyte Injury in Vitro (다형핵구에 의한 폐포세포 손상에 Histamine이 미치는 영향)

  • Kim, Young-Kyoon;Kwon, Soon-Seog;Kim, Kwan-Hyung;Han, Ki-Don;Moon, Hwa-Sik;Sang, Jeong-Sup;Park, Sung-Hak
    • Tuberculosis and Respiratory Diseases
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    • v.39 no.3
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    • pp.228-235
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    • 1992
  • Background: Although polymorphonuclear leukocytes (PMN) are important in protecting the airways and alveolar surfaces, there is evidence that they can also injure the lung while exercising their defensive role. However it has been unclear whether PMN-induced pneumocyte injury is mediated by their direct cytotoxic effect on target cells or by PMN-derived cytotoxic mediators. On the other hand histamine was known not only to act as an important chemical mediator participated in the pathogenesis of some atotic and allegic disorders, but also to have an inhibitory effect on normal PMN functions. Method: To study the mechanism by which PMN induce pneumocyte injury, we cocultured PMN from four healthy nonsmokers or their PMN-derived supernatants (PMN-SPN) with monolayers of $^{51}Cr$-labeled human A549 pneumocytes and compared PMN-and PMN-SPN-mediated pneumocyte injuries measured by $^{51}Cr$ release assay. We also compared the effects of histamine on each pneumocyte injury. Results: 1) PMN-SPN showed more injurious effect on A549 pneumocytes than that of PMN itself regardless histamine pretreatment of PMN. 2) Pneumocyte injury by PMN with histamine pretreatment was increased or decreased compared with that by PMN without histamine pretreatment, according to histamine concentrations, and PMN stimulating agents and their concentrations. 3) Pneumocyte injury by PMN-SPN with histamine pretreatment tended to be decreased compared with that by PMN-SPN without histamine pretreatment. Conclusion: Our results suggest that PMN-SPN may play more important role in mediating pneumocyte injury than PMN itself and that histamine may partially play a protective role on PMN-induced pneumocyte injury. Alternatively we conclude that the effects of histamine on PMN-induced pneumocyte injury may be affected by microenvironment in vivo.

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A study of the effects of PDGF-BB on the characteristics of bone stromal and periodontal ligament cells (혈소판유래성장인자-BB가 골간질세포와 치주인대세포의 성상에 미치는 영향)

  • Kwon, Young-Hyuk;Park, Joon-Bong
    • Journal of Periodontal and Implant Science
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    • v.26 no.4
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    • pp.949-965
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    • 1996
  • The main goal of periodontal therapy is to restore the lost periodontal tissue and establish the attachment appratus. Current acceptable therapeutic techniques are included : removal of diseased soft tissue, demineralization of exposed root surface, using the barrier membrane for preventing the downgrowth of gingival epithelial cell, insertion of graft materials as a scaffolding action, and biological mediators for promoting the cell activity. The latest concept one among them has been studied which based on the knowledge of cellular biology of destructed tissue. Platelet-derived growth factor(PDGF) is one of the polypeptide growth factor which have been reported as a biological mediator to regulate activities of wound healing progress including cell proliferation, migration, and metabolism. The purposes of this study is to evaluate the influences of the PDGF as biological mediator to periodontal ligament and bone marrow cell. Both right and left maxillary first molar were extracted from rat which had treated with 0.4% ${\beta}-Aminopropionitril$ for 5 days, and feeded until designed date to sacrifice under anesthesisa. Periodontal ligament were removed from the extracted socket of the rat, and cultured with Dulbecco's Modified Essential Medium(DMEM) contained with 10% Fetal Bovine Serum, 100U/ml penicillin, $100{\mu}g/ml$ streptomycin, $0.5{\mu}g/ml$ amphotericin-B. Bone marrow cell were culture from bone marrow suspension with which washed out from femur with same medium. The study was performed to evaluate the effect of PDGF to periodontal ligament and bone cell, cell proliferation rate, total protein synthesis, and alkaline phosphatase activity of rat periodontal ligament(PDL) cell and bone stromal(RBS) cell in vitro. The effects of growth factors on both cells were measured at 3, 5th day after cell culture with (control group) or without growth factors(experimental group). The results were as follows: 1. The tendency of cell proliferation under the influence of PDGF showed more rapid proliferation pattern than control at 3 and 5 days after inoculation. 2. The activity of Alkaline phosphatase revealed 14, 80% increased respectively at 3, 5 days culture than control group. Measurements of ALPase levels indicated that PDL cells had significantly higher activity when compared with that of co-culture groups and GF only(P<0.05). And, ALPase activity in 10 days was higher than that of 7 days(P<0.05). 3. The tendency of formation of the mineralized nodule were observed dose-depend pattern of PDL cells. There was statistically significant difference among group l(PDL 100%), 2(PDL 70%:GF 30%), and 3(PDL 50%:GF 50%)(P<0.01). But, there was no difference among group 3, 4(PDL 30%:GF 70%), and 5(GF 100%). 4. Also, the number of nodule was greater in co-culture of PDL 70% and GF 30% than in culture of PDL 70%(P<0.05). From the above results, it is assumed that the PDGF on PDL cells and RMB cell culture. GF stimulates the cell growth, which is not that of PDL cells but GF. And, the activity of ALPase depends on the ratio of PDL cells, and ALPase may relate to the initial phase of nodule formation. Also, it is thought that the calcified nodule formation principally depends on PDL cells, is inhibited by GF, and affected by cell density. In conclusion, platelet-derived growth factor can promote rapid osteogenesis during early stage of periodontal tissue regeneration.

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The effect of silane treatment timing and saliva contamination on shear bond strength of resin cement to porcelain (Silane의 처리시기와 타액오염이 도재-레진 시멘트의 전단 결합강도에 미치는 영향)

  • Ro, Young-Seon;Ryu, Jae-Jun;Suh, Kyu-Won
    • The Journal of Korean Academy of Prosthodontics
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    • v.47 no.1
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    • pp.61-69
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    • 2009
  • Statement of problem: Porcelain veneers have become a popular treatment modality for aesthetic anterior prosthesis. Fitting porcelain veneers in the mouth usually involve a try-in appointment, which frequently results in salivary contamination of fitting surfaces. Purpose: An in vitro study was carried out to investigate the effect of silane treatment timing and saliva contamination on the resin bond strength to porcelain veneer surface. Material and methods: Cylindrical test specimens (n=360) and rectangular test specimens (n=5) were prepared for shear bond test and contact angle analysis. Whole cylindrical specimens divided into 20 groups, each of which received a different surface treatment and/or storage condition. The composite resin cement stubs were light-polymerized onto porcelain adherends. The shear bond strengths of cemented stubs were measured after dry storage and thermocycling (3,000 cycles) between 5 and $55^{\circ}C$. The silane and their reactions were chemically monitored by using Fourier Transform Infrared Spectroscopy analysis (FTIR) and contact angle analysis. One-way analysis of variance (ANOVA) and Dunnett's multiple comparison were used to analyze the data. Results: FT-IR analysis showed that salivary contamination and silane treatment timing did not affect the surface interactions of silane. Observed water contact angles were lower on the saliva contaminated porcelain surface and the addition of 37% phosphoric acid for 20 seconds on saliva contaminated porcelain increased the degree of contact angle. Silane applied to the porcelain, a few days before cementation, resulted in increasing the bond strength after thermocycling. Conclusion: Within the limitation of this study, it can be concluded that it would be better to protect porcelain prosthesis before saliva contamination with silane treatment and to clean the contaminated surface by use of phosphoric acid.

Genetic and Physiological Discrepancies from Isolates of Sclerotinia homoeocarpa causing Zoysiagrass Dollar Spot Disease (한국잔디에 발생하는 동전마름병 원인균의 유전 및 생리적 특성차이)

  • Park, Dae-Sup;Kim, Kyung-Duck;Kihl, Joon-Yeong;Pyee, Jae-Ho
    • Asian Journal of Turfgrass Science
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    • v.20 no.1
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    • pp.65-76
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    • 2006
  • Scz1, an isolate of Sclerotinia homoeocarpa, was recently reported as a novel pathogen responsible for dollar spot disease in Zoysiagrass, a warm season turfgrass. Scz1 possessed different characteristics on mycelial pigment, mycelial affinity and host pathogenecity compared to those of Scb1, a typical isolate, obtained from creeping bentgrass, a cool season turfgrass. In this study, only three isolates, Scz1, Scz2(another analogous isolate of Sclerotinia homoeocarpa from zoysiagrass), and Scb1, were examined at the molecular level using the internal transcribed spacer(ITS) and random amplified polymorphic DNA(RAPD) assays to verify their identification and genetic variation. As a result of ITS assay, partial ITS sequences of three isolates showed 94-97% similarity with a standardized ITS sequence of S. homoeocarpa registered on BLAST. In the analysis of RAPD, range value through similarity matrix was 0.167 between Scz1 and Scb1, 0.139 between Scz2 and Scb1, and 0.713 between Scz1 and Scz2, respectively. Furthermore, tendegram analysis indicated that Scz1 and Scz2, unlike Scb1, were clustered together as accompanying a high genetic similarity. In in vitro fungicide bioassay, $EC_{50}$ value representing the sensitivity degree to propiconazole, a well-known fungicide for dollar spot disease, was 0.012 ${\mu}g/ml$ for Sczl, 0.003 ${\mu}g/ml$ for Scz2, and 0.030 ${\mu}g/ml$ for Scb1. From all data taken, we concluded that both Scz1 and Scz2 belonged to one group of S. homoeocarpa, since they exhibit the same host range and high level of genetic similarity, whereas their chemical competences to a fungicide were different. This study would provide further approach for assessing genetic diversity of S. homoeocarpa isolates as well as characterizing individual isolate against chemical exposure.

Partially purified Toxoplasma gondii antigens by immunoaffinity chromatography (Immunoaffinity chromatography를 이용한 톡소포자충 항원의 부분정제)

  • 안명희;현근희
    • Parasites, Hosts and Diseases
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    • v.35 no.4
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    • pp.251-258
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    • 1997
  • Tachyzoite antigens of Toxoplosnc gondii (RH) were partially purified by immunoaffinity chromatography. The cultivated ToxopLusmc in uiuo (mouse) and in nitro (Hep-2 cell) and peritoneal fluid of T. Bondii infected mice were collected for antigen analy- sis. Tachyzoite antigens collected from infected mouse showed positive bands of 76 kDa, 70 kDa,64 kDa, 53 kDa, 46 kDa, 44 kDa, 41 kDa, 35 kDa, 25 kDa, 18 kDa, and 13 kDa on immunoblot with anti-Toxoplcsmn rabbit sera, and those from infected Hep-2 cells revealed reactive bands of 70 kDa,64 kDa,53 kDa,35 kDa,28 kDa, and 13-10 kDa. After applying to an IgG-Sepharose column, two elusion peaks, E-1 and I-2 fractions, were obtained from both soluble antigen of T. gondii and the peritoneal fluid of infected mice, respectively. Immunoblots of soluble antigen with immunized rabbit sera revealed positive bands of 97 kDa, 63 kDa, 53 kDa and 35 kDa from I-1 fraction and 53 kDa and 35 kDa from I-2. In the case of the eluted peaks from mice peritoneal fluid, E-1 showed protein bands of 84 kDa,76 kDa,53 kDa and 29 kDa bands and 53 kDa and 45 kDa from I-2 on immunoblots. Serum IgG antibody titer of mice immunized with T gonnii tachyzoites was increased on 1 week after booster immunization when analysed by ELISA using crude antigen, while it was elevated on 3 weeks after booster immunization by ELISA using puri- fied antigen.

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ELASTIC CONSTANTS, SHEAR BOND STRENGTH OF TUNNEL RESTORATIVE MATERIALS AND MARGINAL RIDGE STRENGTH OF RESTORED TEETH (터널형 2급와동 충전재의 탄성계수와 전단결합강도 및 수복치의 변연융선 파절강도에 관한 연구)

  • Lee, Ka-Yean;Park, Yeong-Joon;Yang, Kyu-Ho
    • Journal of the korean academy of Pediatric Dentistry
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    • v.23 no.3
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    • pp.746-763
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    • 1996
  • An alternative design to conventional class II cavity preparation for proximal carious lesions is the tunnel preparation. It preserves the marginal ridge intact, thus making it possible to maintain the natural contact relationship with the adjacent tooth and minimize tooth reduction. This in vitro study was purposed to evaluate the effect of the materials' elastic constants and shear-bond strength on the marginal ridge fracture resistance of teeth restored by the tunnel technique, and to find the materials of choice for tunnel restorations. $Resinomer^{(R)}$, $Ketac-silver^{(R)}$, $Miracle-Mix^{(R)}$, and Tytin were used as restorative material. The elastic constants of each restorative material were evaluated by ultrasonic pulse measurement. Young's modulus and bulk modulus of the restorative materials were evaluated in three specimens for each material type. The shear-bond strength of the restorative materials to the dentin surface was measured after thermocycling 400 times between 6 and $60^{\circ}C$, using ten specimens for each material type. For measuring marginal ridge strength, 60 sound extracted molar teeth were distributed into six groups by size. Sound molar teeth were used as a Control group and unfilled prepared teeth were grouped as Unrestored. Another four groups were named Resinomer group, Ketac-Silver group, Miracle Mix group, and Tytin group by type of restorative material. Tunnel cavity preparation was done with ' 1/2, 2, and 4 round burs in sequence. Initial access to proximal surface was made through an occlusal access preparation started at least 2mm from the marginal ridge, and the proximal opening was formed about 2.5mm below the marginal ridge. After restoration and thermocycling, marginal ridge strength was measured using a universal testing machine. The results were as follows: 1. The Young's modulus of $Tytin^{(R)}$ was 63.95 GPa, followed by $Ketac-Silver^{(R)}$ 27.60 GPa, $Miracle-mix^{(R)}$ 18.48 GPa, and $Resinomer^{(R)}$ 10.74 GPa showing significant differences between the groups(P<0.05). The bulk modulus of the materials showed the same order as Young's modulus. The value of $Tytin^{(R)}$ showed 59.57 GPa indicating that it will deform less than other materials under the same stress. It was followed by $Ketac-Silver^{(R)}$ 23.57 GPa, Miracle $Mix^{(R)}$ 12.50 GPa, and $Resinomer^{(R)}$ 11.60 GPa. 2. The Resinomer group had a shear-bond strength of 7.41 MPa which was significantly higher than those of the Ketac-Silver group (1.80 MPa) and the Miracle Mix group (2.84 MPa) (P<0.01). All the specimens of Tytin group detatched from the dentin surface during thermocycling. 3. The mean marginal ridge strength of the Unrestored group(46.14 kgf) was significantly lower than that of the Control group (84.24 kgf) (P<0.01). The marginal ridge strength of teeth restored by the tunnel technique was, in order, Ketac-Silver group 74.06 kgf, Miracle Mix group 73.36 kgf, Resinomer group 63.47 kgf, and Tytin group 58.76 kgf. The Ketac-Silver, Miracle Mix, and Resinomer groups showed no significant difference with the Control group (P>0.05), but the Tytin group showed significantly lower strength compared to the Control group(P<0.05). The results showed that the marginal ridge strength of the teeth restored by the tunnel technique was not significantly lower than that of sound teeth. They also demonstrated that the bonding strength of the restorative material to the tooth surface should be high and the modulus of elasticity should not be lower than that of the tooth in order to restore the marginal ridge strength to its natural condition.

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Effect of Cinnamomum camphora Leaf Fractions on Insulin Action (3T3-L1 지방세포에서 녹나무 잎 추출분획물이 인슐린작용에 미치는 효과)

  • Ko, Byoung-Seob;Lee, Mi-Young;Kim, Ho-Kyoung;Chun, Jin-Mi;Choi, Soo-Bong;Jun, Dong-Wha;Jang, Jin-Sun;Park, Sunmin
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.9
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    • pp.1336-1343
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    • 2005
  • In the present study, we screened candidates for enhancing insulin action and secretion from Cinnamomum camphora (CC) fractions, in 3T3-L1 adipocytes and Min6 cells by investigating insulin- stimulated glucose uptake and glucose-stimulated insulin secretion, respectively. CC were extracted by $70\%$ ethanol followed by XAD-4 column chromatography with serial mixture solvents of methanol and water, and the fractional extractions were utilized for determining insulin action and secretion, and $\alpha$-glucoamylase suppressing activity, A significant insulin-stimulated glucose uptake was observed in 3T3-L1 adipocytes, giving 0.5 or $5{\mu}g/mL$ of $40\%\;and\;60\%$ methanol fractions plus 0.2 nM insulin, compared to the treatment of DMSO plus 0.2 nM insulin. The treatments of $40\%\;and\;60\%$ methanol fractions plus 0.2 nM insulin reached the glucose uptake of 10 nM insulin treatment. The $40\%$ methanol fraction increased triglyceride accumulation by stimulating differentiation and triglyceride synthesis similar to pioglitazone, PPAR-$\gamma$ agonist. No inhibition of $\alpha$-glucoamylase activity of CC fractions was observed. They did not modulate the insulin secretion capacity In either low or high glucose media. These results suggest that $40\%$ methanol fraction contains a potential insulin sensitizer to have a similar function of PPAR-$\gamma$ agonist. Crude CC extract may improve glucose utilization by enhancing insulin-stimulated glucose uptake without elevating glucose stimulated insulin secretion.