• Title/Summary/Keyword: in vitro Metabolism

Search Result 497, Processing Time 0.023 seconds

Metabolism Activity of Bifidobacterium spp. by D.Ps of Konjac Glucomannan Hydrolysates (Konjac Glucomannan 가수분해 올리고당의 중합도별 Bifidobacterium spp.에 대한 대사활성)

  • 최준영;박귀근
    • Journal of the Korean Society of Food Science and Nutrition
    • /
    • v.33 no.7
    • /
    • pp.1186-1191
    • /
    • 2004
  • Bacillus sp. $\beta$-mannanase was purified by DEAE-sephadex ion exchange column chromatography. The partially purified P-mannanase exhibited maximum activity at pH 6.0 and 5$0^{\circ}C$, and was stable at a pH range of 5.5 to 7.0, and at temperature between 30 to 5$0^{\circ}C$. Konjac glucomannan was hydrolyzed by the purified $\beta$-mannanase, and then hydrolysates separated by 1st activated carbon column chromatography and 2nd sephadex G-25 gel filtration. The main hydrolysates were composed of D.P 5 and 7 glucomannooligosaccharides by TLC and FACE method. To investigate the effects of guar gum glucomannooligosaccharides on the in vitro growth of B. longum, B. bifidum, B. infantis, B. adolescentis, B. animalis, and B. breve, Bifidobacterium spp. were cultivated individually on the modified-MRS medium containing carbon SOUTce such as D.P 5, and D.P 7 glucomannooligosaccharides, respectively. B. longum grew up 4.6-fold and 5.3-fold more effectively by the replacement of D.P 5 and 7 glucomannooligosaccharides as the carbon source in a comparasion of standard MRS. Also, B. breve and B. animalis slightly grew up by the treatment of D.P 5 glucomannooligosaccharide.

Metabolic Study on C29-Brassinosteroids in Young Rice Plants (벼 유식물을 이용한 C29-Brassinosteroids의 대사)

  • Won, So-Yun;Joo, Se-Hwan;Kim, Seong-Ki
    • Journal of Plant Biotechnology
    • /
    • v.34 no.3
    • /
    • pp.243-251
    • /
    • 2007
  • It has been recently demonstrated the presence of not only $C_{28}-BRs$ biosysnthesis, but also $C_{27}-$ and $C_{29}-BRs$ biosynthesis in plants, suggesting that BRs biosynthesis are complicatedly connected to produce biologically active BR (s). This prompted us to investigation of metabolism of a $C_{29}-BR$, 28-homoCS in seedlings of rice from which $C_{29}-BRs$ such as 28-homoTE and 28-homoTY have been identified. In vitro enzyme conversion study using a crude enzyme solution prepared from rice seedlings revealed that 28-homoCS is converted into both CS and 26-nor-28-homoCS, but their reversed reaction did not occur. This indicated that 28-homoCS is biosynthetically converted into more biologically active $C_{28}-BR$, CS by C-28 demethylation and biodegraded into 26-nor-28-homoCS by C-26 demethylation. Next, bio-conversion of 28-homoCS to 28-homoBL was examined by the same enzyme solution. No 28-homoBL as a metabolite of 28-homoCS was detected, meaning that biosynthetic reaction for 28-homoCS to 28-homoBL is not contained, and main connection of $C_{28}-BRs$ and $C_{29}-BRs$ biosynthesis is between CS and 28-homoCS in the rice seedling. This study is the first demonstrated that $C_{29}-BRs$ and $C_{28}-BRs$ bionsynthetic pathways are connected, and that $C_{29}-BRs$ biosynthetic pathway is an alternative biosynthetic pathway to produce more biologically active $C_{28}-BR$, CS in plant.

A Study on the Expression of Glycosaminoglycans in the Experimental Tooth Movement of Rat and in Cultured Periodontal Ligament Cells (실험적 치아이동시 glycosaminoglycan의 발현에 관한 연구)

  • Lee, Kyung-Hwan;Lee, Jong-Jin;Kang, Kyung-hwa;Kim, Eun-Cheol;Kim, Sang-Cheol
    • The korean journal of orthodontics
    • /
    • v.31 no.4 s.87
    • /
    • pp.447-458
    • /
    • 2001
  • The purpose of this study was to evaluate 1) in vivo, the expression of chondroitin 4-sulfate (CH-4S), a structural element of glycosaminoglycans(GAGs), in periodontal tissue during the experimental movement of rat incisors, by labelled streptavidine biotin immunohistochemical staining for CH-4S, 2) In vitro, the expression of CH-4S in cultured human periodontal ligament(PDL) cells supplemented with 10ng/ml of $TGF-{\beta}_1$, 20ng/ml of PDGF-BB, 1ng/ml $TNF-\alpha$, or $1{\mu}g/ml$ LPS by western blot analysis. The results of this study were as follows ; 1. The expression of CH-4S was stronger in pulp, PDL, osteoblasts, osteoclasts and osteocytes in experimental group than in control group, but was rare in dentin, and cementum of experimental groups, regardless of the duration of force application, which was not different from that of control group. 2. In experimental group, the expression of CH-4S in pulp began to increase at 1 day after force application and got to the highest degree at 7 days. After 14 days, the expression in CH-4S immunoreactivity was decreased, and became similar to that of control group at 28 days. 3. The expression of CH-4S in PDL was noted in adjacent to alveolar bone. PDL showed higher intensity of immunolabelling after 1 day of orthodontic tooth movement. And the expression was more stronger in the tension side than that of pressure side of PDL at 1 day, but more stronger in the pressure side than that of tension side of PDL at 4 days. After 7 days, a decrease in CH-4S expression was observed. 4. The expression of CH-4S in alveolar bone got to the highest degree at 4 days, and At 7 days, a decrease in CH-4S expression was observed. 5. PDGF-BB notably raised the expression of CH-4S in the PDL cells at 3 days of cultivation 6. The expression of CH-4S of PDL cells was decreased with the application of $TNF-\alpha$ at 1 day. 7. Admixture of $TGF-{\beta}_1$ and PDGF-BB got more expression of CH-4S in PDL as compared to only $TGF-{\beta}1$ or PDGF-BB. A similar decrease of the expression of CH-4S was observed in the case of application of LPS or $TNF-\alpha$.

  • PDF

In Vivo Image and Biodistribution of $^{123}I$-15-(p-iodophenyl)-3-R, S-methylpentadecanoic acid (BMIPP) in Liposarcoma Bearing Nude Mice (지방육종형성 동물모델에서 $^{123}I$-15-(p-iodophenyl)-3-R, S-methylpentadecanoic acid (BMIPP)의 생체분포와 생체영상)

  • Lee, Tae-Sup;Suh, Yong-Sup;Choi, Chang-Woon;Woo, Kwang-Sun;Chung, Wee-Sup;Lim, Soo-Jung;Lim, Sang-Moo;Awh, Ok-Doo
    • The Korean Journal of Nuclear Medicine
    • /
    • v.35 no.5
    • /
    • pp.324-333
    • /
    • 2001
  • Purpose: $^{123}I$-labeled fatty acids have been used in the evaluation of regional myocardial energy metabolism. This study aimed to evaluate the usefulness of $^{123}I$-BMIPP as a liposarcoma-imaging agent. Materials and Methods: We compared in vitro uptakes between liposarcoma(SW872) and glioma(9L) cell lines, and examined biodistribution and in vivo images of $^{123}I$-BMIPP in liposarcoma-bearing nude mice. Cold-BMIPP was labeled with $^{123}I\;using\;Cu^{2+}$ as catalyst. After purification by Sep-pak, radiochemical purity was determined by TLC. We compared cellular uptake between glioma and liposarcoma after incubation of 5, 10, 15, 30, 60, 120, and 180 mins with culture medium containing $^{123}I$-BMIPP. The difference in biodistribution was determined between non-feeding (water only) group for 18 hr and feeding group in normal mice (n=6/group) at 0.5, 2, and 24 hr. In liposarcoma-hearing nude mice model, liposarcoma, SW872, ceil lines were injected subcutaneously into the felt thigh of nude mice. The biodistribution of $^{123}I$-BMIPP was evaluated at 0.5, 2, and 24 hr (n:5 / group) and in vivo Image of $^{123}I$-BMIPP was obtained with gamma camera at 2 and 24 hr in liposarcoma-hearing nude mice. Results: Radiolabeling yield and radiochemical purity were 95% and above 99%, respectively. SW872 cell line showed more increased uptake than 9L with 1.5 times at 180 mins. The clearance of $^{123}I$-BMIPP in various tissues was more delayed in the non-feeding group than in the feeding group, especially at delayed time (24 hr) in normal mice, and the major excreting organ was the gastrointestinal tract. In liposarcoma-bearing nude mice, tumor/blood ratio of $^{123}I$-BMIPP was 0.94, 0.75, and 1.38 and tumor/muscle ratio was 0.66, 1.53, and 1.11 at 0.5, 2, and 24hr, respectively. $^{123}I$-BMIPP was selectively localized in liposarcoma at 24 hr image. Conclusions: These results suggest that $^{123}I$-BMIPP can be used as a liposarcoma-imaging agent.

  • PDF

Changes of Arachidonic Acid Metabolites in Silica-Exposed Alveolar Macrophage of Rats (유리규산분진에 폭로된 흰쥐의 폐포대식세포에 있어 아라키돈산 대사산물의 변화)

  • Lim, Young;Yun, Im-Goung
    • Tuberculosis and Respiratory Diseases
    • /
    • v.39 no.4
    • /
    • pp.304-309
    • /
    • 1992
  • Background: The alveolar macrophage may metabolize arachidonic acid through cyclooxygenase- and lipoxygenase- catalyzed pathways to produce a variety of metabolites of arachidonic acid. The production of these metabolites of arachidonic acid may enhance the defensive ability of the challenged lung. However, continued stimulation with the consequent production of proinflammtory metabolites of arachidonic acid, may ultimately enhance the disease process by contributing to chronic bronchoconstriction, fibrosis, and the persistent release of toxic oxygen species. Silicosis is an example of a disease process resulting from chronic exposure of the lung to foreign particles. This study was carried out to evaluate the changes of arachidonic acid metabolites from macrophages in experimental silicosis. Methods: We measured $PGE_2$, and $LTB_4$ in cultured macrophages taken from rats by radioimmunoassay at 24 and 48 hours after stimulation by silica dust, natural carbon dust, lipopolysaccharide, calcium ionophore (A23187) and medium (RPMI) as a control. For the experimental silicosis, 50 mg silica in 0.5 ml saline was administered intratracheally into the rat and grown to 20 weeks and measured $PGE_2$, and $LTB_4$ in the cultured macrophages lavaged from that rat. The used stimulants were the same as above. Results: 1) The amount of $PGE_2$ in the cultred macrophages from normal rat was significantly decreased in the group which was stimulated with silica dust for 48 hours compare with control non-stimulated group. 2) In the experimental silicosis group, $PGE_2$, release in cultured macrophages after 48 hours incubation with silica and natural carbon dust tended to be lower than those of non-stimulated group. 3) There were marked changes of $LTB_4$ in the groups of normal rats which were incubated with silica for 24, 48 hours and natural carbon for 48 hours compared with non-stimulated group. 4) In the experimental silicosis group, the release of $LTB_4$ was significantly increased macrophages cultured with silica and natural carbon dust after 24 and 48 hours incubation compared with non-stimulated group. Conclusion: The results of these studies suggest that the in vitro exposure of rat alveolar macrophge to silica and coal dust results in an alteration in alveolar macrophage metabolism of arachidonic acid that may promote an inflammatory reaction in lung tissue.

  • PDF

Pomegranate Extract Improves Menopausal Syndrome in Ovariectomized Rats (난소 적출 동물모델에서 고흥산 석류 농축액의 갱년기 증상 개선 효과)

  • Wee, Ji-Hyang;Jung, Hyun Jung;Jung, Kyung Ok;Sung, Hea Mi;Shin, Yu-Rim;Park, Ju-Hyun;Seo, Hyeon-Young;Lim, Jung-Min;Chae, Han-Jeong;Lee, Ki Yung
    • Journal of the Korean Society of Food Science and Nutrition
    • /
    • v.44 no.4
    • /
    • pp.506-515
    • /
    • 2015
  • In this study, the effect of Goheung pomegranate extract on postmenopausal syndrome was evaluated both in vitro and in vivo in ovariectomized Sprague-Dawly (SD) rats. Sixty female SD rats were divided into six groups: sham, sham operation and distilled water; OVX, ovariectomized and distilled water; PE1, ovariectomized and pomegranate concentrate (0.75 mL/twice/d); PE2, ovariectomized and pomegranate concentrate (1.5 mL/twice/d); PE3, ovariectomized and pomegranate concentrate (2.2 mL/twice/d); and CE, ovariectomized and commercial pomegranate concentrate (2.2 mL/twice/d). Percent bone volume (bone volume/tissue volume) and trabecular thickness (Tb.Th) improved in a dose-independent manner in PE1, 2, and 3. Especially, bone mineral density was significantly improved in PE3 (P<0.05) compared to OVX. Pomegranate extract reduced body weight and visceral fat mass. High density lipoprotein cholesterol (HDL-C) level slightly increased in a dose-independent manner in the experimental group. In addition, HDL-C/total cholesterol level of PE3 significantly increased (P<0.05) compared with OVX. These results show that pomegranate concentrate improved blood lipid levels and bone metabolism in ovariectomized rats. Therefore, Goheung pomegranate concentrates are expected to improve cardiovascular and bone-related diseases in menopausal women.

Identification of Matrix Mineralization-Related Genes in Human Periodontal Ligament Cells Using cDNA Microarray (cDNA microarray에 의한 치주인대세포의 광물화 결절형성에 관여하는 유전자들의 분석)

  • Shin, Jae-Hee;Park, Jin-Woo;Yeo, Shin-Il;Noh, Woo-Chang;Kim, Moon-Kyu;Kim, Jung-Chul;Suh, Jo-Young
    • Journal of Periodontal and Implant Science
    • /
    • v.37 no.sup2
    • /
    • pp.447-463
    • /
    • 2007
  • Periodontal ligament (PDL) cells have been known as multipotential cells, and as playing an important rolesin periodontal regeneration. The PDL cells are composed of heterogeneous cell populations which have the capacity to differentiate into either cementoblasts or osteoblasts, depending on needs and conditions. Therefore, PDL cells have the capacity to produce mineralized nodules in vitro in mineralization medium which include ascorbic acid, ${\beta}$-glycerophosphate and dexamethasone. In spite of these well-known osteoblast like properties of PDL cells, very little is known about the molecules involved in the formation of the mineralized nodules in the PDL cells. In the present study, we analysed gene-expression profiles during the mineralization process of cultured PDL cells by means of a cDNA microarray consisting of 3063 genes. Nodules of mineralized matrix were strongly stained with alizarin red S on the PDL cells cultured in the media with mineralization supplements. Among 3,063 genes analyzed, 35 were up-regulated more than two-fold at one or more time points in cells that developed matrix mineralization nodules, and 38 were down-regulated to less than half their normal level of expression. In accord with the morphological change we observed, several genes related to calcium-related or mineral metabolism were induced in PDL cells during osteogenesis, such as IGF-II and IGFBP-2. Proteogycan 1, fibulin-5, keratin 5, ,${\beta}$-actin, ${\alpha}$-smooth muscle actin and capping protein, and cytoskeleton and extracellular matrix proteins were up-regulated during mineralization. Several genes encoding proteins related to apoptosis weredifferentially expressed in PDL cells cultured in the medium containing mineralization supplements. Dkk-I and Nip3, which are apoptosis-inducing agents, were up-regulated, and Btf and TAXlBP1, which have an anti-apoptosis activity, were down-regulated during mineralization. Also periostin and S100 calciumbinding protein A4 were down-regulated during mineralization.

The effect of admixture of vitamin $D_3$ and dexamethasone on the activity of osteoblastic cells (Vitamin $D_3$와 Dexamethasone의 복합 투여가 골모세포에 미치는 영향에 관한 연구)

  • Lim, Na-Won;Park, Young Joo;Kim, Sang-Cheol
    • The korean journal of orthodontics
    • /
    • v.29 no.3 s.74
    • /
    • pp.383-397
    • /
    • 1999
  • Bone is a dynamic tissue which is constantly remodelled by subsequent cycles of bone resorption and formation. Glucocorticoid and vitamine $D_3$ are known as regulating substances in bone metabolism. In vitro experiments using bone tissue, it was suggested that glucocorticoid inhibits bone resorption, whereas the effect of glucocorticoid on bone formation are complex- increasing or decreasing effect. The active form of vitamin $D_3$, 1,25-dihydroxycholecalciferol[1.25-$(OH)_2D_3$], has been reported to stimulate osteoblastic activities including the production of ALP, type I collagen, and osteoclacin. The purpose of this study was to evaluate the effect of admixture of vitamin $D_3$ and dexamethasone, one of glucocorticoids, on osteoblastic cell line(MC3T3-E1). Alkaline phosphatase(ALP) and MTT assay were conducted in the cultivated cells with 1, 10, 100nM/ml of 1,25-$(OH)_2D_3$ and/or 10nM/ml, 100nM/ml, $1{\mu}M/ml$ of dexamethasone. The observed results were as follows. 1. The activity of osteoblastic cells with $1{\mu}M/ml$ of dexamethasone was significantly increased at 1-day cultivation with comparison to control group, but was decreased afterwards. But the activity of ALP was greatest in $1{\mu}M/ml$ of dexamethasone and increased with time lapsed. 2. The activity of osteoblastic cells with vitamin $D_3$ was significantly increased dose-dependently at 1-day cultivation, but was significantly decreased in l00nM/.ml at 2-day cultivation, and was a little increased again at 3-day cultivation. The activity of ALP was increased in 10nM/ml or 100nM/ml at 2-day or 3-day cultivation, and was greatest in 100nM/ml at 3-day cultivation. 3. In case of admixture of dexamethasone and vitamin $D_3$, the cellular activity was decreased in any concentration of vitamin $D_3$ at 2-day cultivation, but was increased again at 3-day cultivation, which was greater than that in control or dexamethasone only group. The activity of ALP was decreased at 1-day cultivation, but was increased in the admixture of 10nM/ml or 100nM/ml of dexamethasone with 100nM/ml of vitamin $D_3$ at 2-day cultivation, and was again decreased at 3-day cultivation.

  • PDF

Immunostimulatory and Anti-Obesity Activity of Lonicera insularis Nakai Extracts in Mouse Macrophages RAW264.7 Cells and Mouse Adipocytes 3T3-L1 Cells (섬괴불나무(Lonicera insularis Nakai) 추출물의 면역자극 및 항비만 활성)

  • Yu, Ju Hyeong;Yeo, Joo Ho;Choi, Min Yeong;Lee, Jae Won;Geum, Na Gyeong;An, Mi-Yun;Jeong, Jin Boo
    • Korean Journal of Plant Resources
    • /
    • v.35 no.4
    • /
    • pp.417-427
    • /
    • 2022
  • In this study, we investigated in vitro immuno-stimulatory and anti-obesity activity of fruit (LIF), leaves (LIL) and stems (LIS) from Lonicera insularis Nakai in mouse macrophages RAW264.7 cells and mouse pre-adipocytes 3T3-L1 cells. LIF, LIL and LIS increased the production of immunostimulatory factors such as nitric oxide (NO), inducible nitric oxide synthase (iNOS), interleukin-1β (IL-1β), cyclooxygenase-2 (COX-2), tumor necrosis factor-α (TNF-α) and activated phagocytosis in RAW264.7 cells. Inhibition of toll-like receptor 2/4 (TLR2/4) partly blocked LIF, LIL and LIS mediated production of immunostimulatory factors. In addition, inhibition of mitogen-activated protein kinases (MAPK) signaling attenuated the production of immunostimulatory factors induced by LIF, LIL and LIS. Based on these results of this study, LIF, LIL and LIS is thought to activate macrophages the production of immunostimulatory factors and phagocytosis through toll-like receptor 2/4 (TLR2/4) and MAPKs signaling pathway. In anti-obesity study, LIF reduced the lipid accumulation in 3T3-L1 cells. LIF increased the protein phosphorylation expressions such as AMP-activated protein kinase (AMPK), hormone sensitive lipase (HSL), adipose triglyceride lipase (ATGL) related to the lipolysis of the adipocytes. In addition, LIF increased the expression of proteins involved in energy metabolism and brown adipose tissues differentiation such as peroxisome proliferator-activated receptor gamma coativator 1α (PGC-1α) and PR domain-containing16 (PRDM16). These results suggest that LIF is involved in lipid accumulation inhibition through expressing the proteins such as lipolysis and differentiation of white adipocytes to brown adipocytes.

Effect of Buan Mulberry on Metabolic Improvement in Streptozotocin-Induced Diabetic Rats (Streptozotocin으로 유발된 당뇨쥐의 신진대사 증진에 대한 부안 오디의 효과)

  • Lee, Ju-Taek;Ryu, Jae-Sung;Kwak, Dong-Hoon;Park, Yun-Jum;Kang, Seong-Sun;Kim, Pyoung-Jun;Hwang, Kyung-A;Choo, Young-Kug
    • Journal of Life Science
    • /
    • v.20 no.12
    • /
    • pp.1750-1757
    • /
    • 2010
  • This study was designed to evaluate the anti-diabetic effects of Buan mulberries by using an insulin-dependent diabetes mellitus animal model. Several studies have shown that mulberries have metabolism-improving, antioxidant, and lipid-lowering properties in rats with streptozotocin (STZ)-induced diabetes. In this study, Sprague-Dawley male rats were randomly assigned to 1 normal control group and 5 STZ-induced diabetes groups: rats that had STZ-induced diabetes and did not receive any agents (diabetic group; negative control), rats that had STZ-induced diabetes and received insulin (insulin group; positive control), rats that had STZ-induced diabetes and received 0.5% mulberry extract (0.5% mulberry group), rats that had STZ-induced diabetes and received 1.0% mulberry extract (1.0% mulberry group), and rats that had STZ-induced diabetes and received 2.0% mulberry extract (2.0% mulberry group). Mulberry extracts were administered to the diabetic animals for 4 weeks. The rats that received mulberry extracts showed lower body weights and insulin levels, as well as higher kidney weights, blood glucose levels, urine quantities, and water intake in comparison with the normal controls. Further, the insulin concentrations in the mulberry-fed animals were higher than those in the diabetic group, and the kidney weights, blood glucose levels, urine quantities, and water intake in the mulberry-fed animals were lower than the corresponding values in the diabetic controls. These results suggest that mulberry may be an effective functional food to prevent diabetes-related complications.