• Title/Summary/Keyword: In vivo와 In vitro

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The Effects of Retinoic Acid and MAPK Inhibitors on Phosphorylation of Smad2/3 Induced by Transforming Growth Factor β1

  • Lee, Sang Hoon;Shin, Ju Hye;Shin, Mi Hwa;Kim, Young Sam;Chung, Kyung Soo;Song, Joo Han;Kim, Song Yee;Kim, Eun Young;Jung, Ji Ye;Kang, Young Ae;Chang, Joon;Park, Moo Suk
    • Tuberculosis and Respiratory Diseases
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    • v.82 no.1
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    • pp.42-52
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    • 2019
  • Background: Transforming growth factor ${\beta}$ (TGF-${\beta}$), retinoic acid (RA), p38 mitogen-activated protein kinase (MAPK), and MEK signaling play critical roles in cell differentiation, proliferation, and apoptosis. We investigated the effect of RA and the role of these signaling molecules on the phosphorylation of Smad2/3 (p-Smad2/3) induced by TGF-${\beta}1$. Methods: A549 epithelial cells and CCD-11Lu fibroblasts were incubated and stimulated with or without all-trans RA (ATRA) and TGF-${\beta}1$ and with MAPK or MEK inhibitors. The levels of p-Smad2/3 were analyzed by western blotting. For animal models, we studied three experimental mouse groups: control, bleomycin, and bleomycin+ATRA group. Changes in histopathology, lung injury score, and levels of TGF-${\beta}1$ and Smad3 were evaluated at 1 and 3 weeks. Results: When A549 cells were pre-stimulated with TGF-${\beta}1$ prior to RA treatment, RA completely inhibited the p-Smad2/3. However, when A549 cells were pre-treated with RA prior to TGF-${\beta}1$ stimulation, RA did not completely suppress the p-Smad2/3. When A549 cells were pre-treated with MAPK inhibitor, TGF-${\beta}1$ failed to phosphorylate Smad2/3. In fibroblasts, p38 MAPK inhibitor suppressed TGF-${\beta}1$-induced p-Smad2. In a bleomycin-induced lung injury mouse model, RA decreased the expression of TGF-${\beta}1$ and Smad3 at 1 and 3 weeks. Conclusion: RA had inhibitory effects on the phosphorylation of Smad induced by TGF-${\beta}1$ in vitro, and RA also decreased the expression of TGF-${\beta}1$ at 1 and 3 weeks in vivo. Furthermore, pre-treatment with a MAPK inhibitor showed a preventative effect on TGF-${\beta}1$/Smad phosphorylation in epithelial cells. As a result, a combination of RA and MAPK inhibitors may suppress the TGF-${\beta}1$-induced lung injury and fibrosis.

Biochemical Assessment of Deer Velvet Antler Extract and its Cytotoxic Effect including Acute Oral Toxicity using an ICR Mice Model (ICR 마우스 모델을 이용한 녹용 추출물의 생화학적 평가 및 급성 경구 독성을 포함한 세포 독성 효과)

  • Ramakrishna Chilakala;Hyeon Jeong Moon;Hwan Lee;Dong-Sung Lee;Sun Hee Cheong
    • Journal of Food Hygiene and Safety
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    • v.38 no.6
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    • pp.430-441
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    • 2023
  • Velvet antler is widely used as a traditional medicine, and numerous studies have demonstrated its tremendous nutritional and medicinal values including immunity-enhancing effects. This study aimed to investigate different deer velvet extracts (Sample 1: raw extract, Sample 2: dried extract, and Sample 3: freeze-dried extract) for proximate composition, uronic acid, sulfated glycosaminoglycan, sialic acid, collagen levels, and chemical components using ultra-performance liquid chromatography-quadrupole-time-of-light mass spectrometry. In addition, we evaluated the cytotoxic effect of the deer velvet extracts on BV2 microglia, HT22 hippocampal cells, HaCaT keratinocytes, and RAW264.7 macrophages using the cell viability MTT assay. Furthermore, we evaluated acute toxicity of the deer velvet extracts at different doses (0, 500, 1000, and 2000 mg/kg) administered orally to both male and female ICR mice for 14 d (five mice per group). After treatment, we evaluated general toxicity, survival rate, body weight changes, mortality, clinical signs, and necropsy findings in the experimental mice based on OECD guidelines. The results suggested that in vitro treatment with the evaluated extracts had no cytotoxic effect in HaCaT keratinocytes cells, whereas Sample-2 had a cytotoxic effect at 500 and 1000 ㎍/mL on HT22 hippocampal cells and RAW264.7 macrophages. Sample 3 was also cytotoxic at concentrations of 500 and 1000 ㎍/mL to RAW264.7 and BV2 microglial cells. However, the mice treated in vivo with the velvet extracts at doses of 500-2000 mg/kg BW showed no clinical signs, mortality, or necropsy findings, indicating that the LD50 is higher than this dosage. These findings indicate that there were no toxicological abnormalities connected with the deer velvet extract treatment in mice. However, further human and animal studies are needed before sufficient safety information is available to justify its use in humans.

Comparison of Flavonoid Contents and Antioxidant Activity of Yuzu (Citrus junos Sieb. ex Tanaka) Based on Harvest Time (산지별 유자의 수확시기에 따른 플라보노이드 함량 및 항산화활성 비교)

  • Moon, So Hyun;Assefa, Awraris Derbie;Ko, Eun Young;Park, Se Won
    • Horticultural Science & Technology
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    • v.33 no.2
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    • pp.283-291
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    • 2015
  • The aim of this study was to evaluate the changes of fruit quality, flavonoid contents and antioxidant activity of Yuzu (Citrus junos Sieb. ex Tanaka) from Go-heung and Jeju according to harvest time. Samples were harvested from August to December on the $1^{st}$ of every month. August and September samples were green colored, whereas November and December samples were yellow. The fruit shape index decreased, changing from globular to elliptical, whereas the $^{\circ}birx$ increased with ripening stage. The yuzu from Jeju was larger than that from Go-heung in each month of cultivation. August samples exhibited the highest amounts of phenolic compounds. In addition, samples from Jeju had higher total phenolic content than those from Go-heung. The content of phenolic compounds decreased with ripening until October and then increased subsequently. Antioxidant activity of the yuzu was evaluated by FRAP and DPPH methods. The antioxidant activity showed a similar trend as total phenolic content. Immature yuzu fruit was found to exhibit the highest amount of flavonoids such as naringin and hesperidin. November and December samples showed almost the same contents of flavonoids. The flavonoid content of yuzu fruit harvested from Jeju was higher than that from Go-heung. Overall, the samples harvested at the early stage, in the month of August, exhibited the highest flavonoid content, phenolic content and antioxidant activity. As the health benefits of these compounds has been demonstrated in various studies, the immature yuzu appears to be preferable for use as a raw material for formulation of pharmaceutical products as well as for functional food production after a proper in-vivo and in-vitro medical tests.

Hemopoietic and Radioprotective Effects of Shenrong Fuzheng Tang(S.F.T.;蔘茸扶正湯) (삼용부정탕(蔘茸扶正湯)의 방사선(放射線) 조사(照射)로 손상(損傷)된 조직(組織) 회부(恢復) 및 조혈촉진(造血促進) 효과(效果))

  • Kim, Jeong-Su;Choe, Seung-Hun;An, Gyu-Seok
    • THE JOURNAL OF KOREAN ORIENTAL ONCOLOGY
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    • v.3 no.1
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    • pp.129-147
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    • 1997
  • Radiotherapy is an irreplaceable method of cancer treatment. But it has several side effects, especially damages to the hemopoietic and Immune system. Therefore radioprotectors are required to treat cancer successfully. A lot of Herbs and Herbal prescriptions are reported to have radioprotective effects. Above all, those to support the healthy energy and strengthen the body resistance are found more effective. This study was performed to evaluate the radioprotective effects of prescription Shenrong Fuzheng Tang(S.F.T.), which consists of 16 kinds of herbs. We investigated proliferation of murine splenocytes, secretion of colony-stimulating-factors(CSFs), immunocompetence after irradiation in-vitro, and Endogenous spleen colony assay, survival assay in-vivo. When splenocytes were cultured with Shenrong Fuzheng $Tang(S.F.T.)(500{\mu}g/ml)$, proliferation was enhanced 5.7 times compared to control cultured with medium alone(p<0.05) and, showed highest proliferation at 4th day after incubation. In order to evaluate stimulation of hemopoiesis of Shenrong Fuzheng Tang(S.F.T.), the supernatant of splenocytes cultured with optimal concentration of Shenrong Fuzheng Tang(S.F.T.) was used to measure CSFs secretion. The result showed enhanced secretion of colony-stimulating-factors (CSFs) compared to control(p<0.05). To evaluate the protective effect of lymphocytes from irradiation, proliferation of splenocytes stimulated by LPS and ConA after incubation with Shenrong Fuzheng Tang(S.F.T.) for 24h Prior to Irradiation$(1{\sim}3\;Gy)$ was measured. The results showed higher proliferation of Shenrong Fuzheng Tang(S.F.T.) treated cells than that of non-treated cells. And percentage increases of irradiated splenocytes per non-irradiated splenocytes were also higher in Shenrong Fuzheng Tang (S.F.T.)-treated cells than control. Endogenous spleen colony assay. to evaluate the protection of hemopoietic cells from irradiation, showed increased number of colonies(p=0.03) in Shenrong Fuzheng Tang(S.F.T.) treated murine spleen$(10.3{\pm}1.9)$ compared to non-treated murine spleen$(3.4{\pm}0.8)$. Survival time of mice irradiated with lethal dose of ${\gamma}-ray(9Gy)$ was prolonged in Shenrong Fuzheng Tang(S.F.T.) treated group prior to irradiation as compared to non-treated group. According to these results we can suggest that prescription Shenrong Fuzheng Tang(S.F.T.) has radioprotective effects and can be used to protect the hemopoietic and immune system from damages of anti-cancer radiotherapy.

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Angiotensin Converting Enzyme Inhibition and Antihypertensive Effects of Phyllostachys pubescens Culm Extracts in Spontaneously Hypertensive Rats (맹종죽 줄기 추출물을 투여한 본태성 고혈압 쥐(SHR)의 ACE 저해 활성 및 혈압 강하 효과)

  • Kim, Jung-Suk;Kim, Mi-Jeong;Park, Min-Hee;Ryu, Bog-Mi;Moon, Gap-Soon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.37 no.1
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    • pp.27-34
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    • 2008
  • This study investigated the anti-hypertensive effect of Phyllostachys pubescens culm extract (PCE) by examining its effects on renal angiotensin-converting enzyme (ACE) inhibition and blood pressure using the spontaneously hypertensive rat (SHR) system. Systolic blood pressure (SBP) and diastolic blood pressure (DBP) were measured weekly for 8 weeks. Also, total antioxidant capacity and protein oxidation of tissues were examined by plasma Trolox Equivalent Antioxidant Capacity assay (TEAC) and hepatic protein carbonyl values, respectively. Twenty male SHR were randomly divided into four groups: PCE50, PCE100, and PCE500 (50, 100, and 500 mg of PCE per kilogram bodyweight daily, respectively), and control group. At week 2, the SBP in all PCE groups appeared to be significantly lower than the control (p<0.05), whereas the DBP were not different until week 4 (p<0.05). At week 8, SBP in the PCE500 was lower by 20% than the control. PCE groups considerably suppressed ACE dose-dependently compared with the control. Plasma TEAC and hepatic protein carbonyl values indicated increased antioxidative activity due to the PCE feed. No adverse effect was observed on the liver of SHR as there was no difference for the GOT and GPT values among the groups. Results of this study suggest that ACE inhibition may be one possible mechanism for the blood pressure lowering effect of PCE; thus, long term consumption of PCE may be beneficial in preventing high blood pressure along with the increased antioxidative status.

Evaluation of Sprouted Barley as a Nutritive Feed Additive for Protaetia brevitarsis and Its Antibacterial Action against Serratia marcescens (흰점박이꽃무지 사료첨가제로서 새싹보리의 곤충병원성 세균에 대한 항균 효과에 관한 연구)

  • Song, Myung Ha;Kim, Nang-Hee;Park, Kwan-Ho;Kim, Eunsun;Kim, Yongsoon
    • Journal of Life Science
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    • v.31 no.5
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    • pp.475-480
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    • 2021
  • Interest in edible insects such as Protaetia brevitarsis has increased rapidly, and several insect producers use these insects in industrialized mass production. However, mass rearing of insects can cause insect diseases. Sprouted barley is a valuable source of nutrients and has antioxidant, antimicrobial, anti-inflammatory, and anti-cancer effects. This study was conducted to investigate the effect of sprouted barley as a feed additive for producing healthy P. brevitarsis larvae. P. brevitarsis larvae were fed feeds with or without sprouted barley, and their body weight and larval period wewe checked weekly. To confirm the antibacterial effects of sprouted barley, in vitro bioassays were performed by counting Serratia marcescens colonies, and in vivo bioassays were performed by determining the survival rate and body weights of the S. marcescens-infected larvae. Larvae fed different feeds were analyzed for their nutrient compositions (i.e., such as proximate composition, minerals, amino acids, and heavy metals). Larvae fed 5% and 10% sprouted barley had maximum weight increases of 19.2% and 23.1%, respectively. Both treatment groups had significantly shorter larval periods than those of the control group. Sprouted barley markedly inhibited the growth of entomopathogenic S. marcescens. Furthermore, larvae fed sprouted barley exhibited higher Cu, Zn, and K levels. Seventeen amino acids were present in larvae fed sprouted barley, of which, tyrosine and glutamic acid were predominant. No heavy metals were detected in any of the investigated groups. Therefore, sprouted barley may be a suitable feed additive for producing high-quality P. brevitarsis larvae.

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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Hypoglycemic Effects of Fruits and Vegetables in Hyperglycemic Rats for Prevention of Type-2 Diabetes (고혈압쥐의 과일과 야채의 섭취에 따른 저혈당 효과)

  • Survay, Nazneen Shaik;Ko, Eun-Young;Upadhyay, Chandrama Prakash;Jang, Mi;Park, Se-Won;Lee, Dong-Ha;Jung, Yi-Sook;Yoon, Do-Young;Hong, Sae-Jin
    • Horticultural Science & Technology
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    • v.28 no.5
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    • pp.850-856
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    • 2010
  • An in vivo oral glucose tolerance test (OGTT) was performed on hyperglycemic male Sprague-Dawley rats to assess the effect of fruits and vegetables ($1g{\cdot}kg^{-1}$ body weight) on blood glucose levels (${\Delta}BGLs$) at different time intervals of 0, 5, 15, 30, 60, 90 and 120 min. The areas under glucose curve (${\Delta}AUCs$) were calculated at 120 min of OGTT by trapezoid method. Total phenolic content (TPC) and anti-oxidant activity (AOA) of fruits and vegetables were assayed in vitro by Folin Ciocalteu and DPPH (2, 2-diphenyl-1-picrylhydrazyl) methods, respectively. At the end of the experiment the correlations among the parameters TPC, AOA and ${\Delta}AUC$ was estimated by Pearson's correlations. Among fruit crops, tangerine, plum, grape and pear and among vegetables, blue leaf mustard, cabbage, chicory, broccoli and others exhibited significant hypoglycemic effects by reducing ${\Delta}BGLs$ with significant ${\Delta}AUC$. The effective ${\Delta}AUC$ ranged from $5548.2{\pm}462.1$ to $3823.3{\pm}282.0mg-min{\cdot}dL^{-1}$. The TPC and AOA ranged from $0.063{\pm}0.00$ to $0.913{\pm}0.14mg{\cdot}g^{-1}$ GAE and $01.05{\pm}0.08$ to $75.46{\pm}0.06%$, respectively. Overall, six fruits and fifteen vegetables exhibited higher TPC and one fruit and four vegetables exhibited higher AOA. There was a better correlation among TPC, AOA and ${\Delta}AUC$ of fruits and TPC & AOA of vegetables. We report that hypoglycemically significant fruits and vegetables investigated in this study have pharmacological importance which reduced ${\Delta}BGLs$ through insulin like activity and AOA in prevention of type-2 diabetes.

EFFECT OF HYDROGEN PEROXIDE CONCENTRATION ON THE WHITENING AND PHYSICAL PROPERTIES OF HYDROXYAPATITE DISCS (Hydrogen Peroxide 농도와 적용시간이 Hydroxyapatite Discs의 미백과 물리적 성질에 미치는 영향)

  • Yang, Yeon-Mi;Lee, Doo-Cheol;Baik, Byeong-Ju;Kim, Jae-Gon;Shin, Jeong-Geun
    • Journal of the korean academy of Pediatric Dentistry
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    • v.34 no.1
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    • pp.1-12
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    • 2007
  • The purpose of this study was to evaluate the effect that various concentration and application time of hydrogen peroxide had on tooth whitening and physical properties. The hydroxyapatite (HA) discs of $12mm({\Phi}){\times}1.2mm(t)$ in dimensions were made by compression $(100kg/cm^2)$ and sintering (at $1350^{\circ}C$ for 2 hours) All specimens were polished sequentially with '240 through '2000 emery paper and one side of each specimen was polished finally with $0.3{\mu}m$ alumina paste. The discs were placed in sterile whole stimulated saliva overnight at $37^{\circ}C$ in order to form an in vitro pellicle layer. Then the discs were rinsed with distilled water and soaked into staining broth at $37^{\circ}C$ for 7 days. These stained specimens were bleached with hydrogen peroxide according to the change of concentration $(3{\sim}30%)$ and application time ($3{\sim}10$ days). The specimens were analyzed with a spectrophotometer, X-ray diffractometer (XRD), scanning electron microscope (SEM), surface roughness tester, microhardness tester and biaxial flexural strength. The results of present study can be summarized as follows : 1. The bleaching effect was increased with the increased concentration and the extended application time of hydrogen peroxide. 2. The surface roughness was significantly increased from the specimen bleached with 15% hydrogen peroxide for 10 days and with 30% for 7 and 10 days respectively (p<0.05). 3. The changes of crystal phase observed by XRD between before and after bleaching weren't shown of any difference, but microporous structure of surface observed by SEM was shown of increase with the increased concentration and the extended application. 4. The biaxial flexural strength was significantly decreased from bleaching of specimen with 30% hydrogen peroxide for 7 and 10 days respectively (p<0.05) 5. Microhardness was significantly decreased from bleaching with 15% hydrogen peroxide for 10 days and with 30% for 3, 7 and 10 days respectively (p<0.05). Although the tooth bleaching effect was greater when the high concentration was applied, further in vivo experiment will be needed to prove it's safety.

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Inhibitory Effects of Lentinus edodes and Rice with Lentinus edodes mycelium on Diabetes and Obesity (In vitro 및 In vivo에서 표고버섯, 표고버섯균사체배양쌀 추출물의 항당뇨와 항비만 작용)

  • Kim, Haeseop;You, Jeheon;Jo, Yeongcheol;Lee, Youngjae;Park, Inbae;Park, Jeongwook;Jung, Myung-A;Kim, Young-Suk;Kim, Sunoh
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.2
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    • pp.175-181
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    • 2013
  • This study investigated the ability of extracts from Lentinus edodes (LE) and rice with Lentinus edodes mycelium (LEM) to inhibit diabetes and obesity. Lipid accumulation significantly decreased by 78% and 74% upon treatment with 300 ${\mu}g/mL$ of LE and LEM, respectively (p<0.01). Cholesteryl ester transfer protein (CETP) inhibition activity increased by 94% and 99% upon treatment with 300 ${\mu}g/mL$ of LE and LEM, respectively. In order to investigate the effect of LE and LEM on diabetes, the inhibition of protein tyrosine phosphate 1B (PTP1B) activity from the LE and LEM extracts at various concentrations (1, 3, 10, 30, 100, 300 ${\mu}g/mL$) was assessed. PTP1B activity by treatment with 10, 30, and 100 ${\mu}g/mL$ of LE, was inhibited at a rate of 7, 9, and 7% respectively. Also, PTP1B activity from treatment with increasing concentration of LEM led to a significant concentration-dependent inhibition of PTP1B activity (p<0.01). LE and LEM were orally administered for 28 days after a high fat diet (HFD). LE and LEM significantly reduced triglyceride, cholesterol, HDL-cholesterol and LDL-cholesterol levels. GOT and GPT were not significantly effected. These results indicate that extracts of LE and rice with LEM have potent activities useful in the treatment of obesity and diabetes mellitus.