Diabetes mellitus is associated with a wide range of pathophysiological in the kidney. This study was designed to examine the effects of high glucose concentration on IGF-I binding and glucose transporters in renal proximal tubule cells. The results were as follows : The binding of $^{125}I-IGF-I$ reached the peak at the 30 minutes and gradually decreased by the time dependent manner. The binding of $^{125}I-IGF-I$ was inhibited by the unlabelled IGF-I($10^{-14}{\sim}10^{-8}M$) in a concentration dependent manner. The relative affinity of IGF-I receptor for IGF-I, IGF-II and insulin exhibited typical type 1 binding(IGF-I > insulin > IGF-II). However IGF-II did not compete for the cultured cell membrane $^{125}I-IGF-I$ binding site at $10^{-14}{\sim}10^{-8}M$. Under optimal conditions, IGF-I binding to the membranes from 5mM and 20mM glucose treated cells was analyzed. It was found that 20mM glucose treated cells exhibited higher binding activity for IGF-I. In order to further substantiate this increase in IGF-I binding sites, we performed affinity-labelling studies. The cross-linked cell membrane subjected to SDS-PAGE; labelled material was detected by autoradiography. 20mM glucose treated cells exhibited higher levels. The initial rate of $methyl-{\alpha}-D-glucopyranoside({\alpha}-MG)$ uptake was significantly lower($74.41{\pm}6.71%$) in monolayers treated with 20mM glucose than those of 5mM glucose. However, 3-O-methyl-D-glucose(3-O-MG) uptake was not affected by glucose concentration in culture media. IGF-I significantly increased ${\alpha}-MG$ uptake in both 5mM and 20mM glucose treated cells. However, 3-O-MG uptake was not affected by IGF-I in both conditions. In conclusion, 20mM glucose increased binding sites of $^{125}I-IGF-I$, inhibited Na/glucose cotransporter activity. But 20mM glucose did not change facilitated glucose transporter.
Heterotrophic activity, total bacteria and salinity were determined seasonally in the estuary of Naktong River over half tidal cycle. Heterotrophic activity was determined by the uptake of [U- $^{14}$ C]glucose. Heterotrophic activity fluctuated with the tides and was decreased as salinity increased. Teh great activity occurred near low ebb tide at all seasons except summer. The main environmental factor affecting hetreotrophic activity was the salinity rather than water temperature in the estuary of Naktong River. In order to estimate the effect of salt, salt was added to estuarine water. Vmax for glucose of salt-added water was 17% and 77% of original estuarine water at station 1 and 2 respectively and slight increase was observed at station 3. Respiration rate and Kt+Sn for glucose of salt-added sample increased at all 3 stations. The increase of the Kt value implies the reduced affinity of bacterial population for glucose. The effects of salinity on the heterotrophic activity were more extensive in the upper region of estuary than at the mouth.
The most efficient method for reducing the organic content of dilute liquid waste is by aerobic-biological treatment. Basically, the organisms responsible for treatment possess the ability to decompose complex organic compounds and to use the energy so liberated for their bodily functions: reproduction, growth, locomotion and so on. That part of organic matter used to produce energy is converted to the essentially stable end products of carbon dioxide, water and ammonia, while the remainder is converted to new cells which can be settled and thus removed from the liquid before the waste is discharged to the receiving body water. Oxygen must be supplied continuously during the aerobic process. In the field of sewage treatment the Warburg respirometer is used mainly for the measurement of the oxygen uptake of samples. In this experiment the Warburg constant volume respirometer was used to determine the oxygen uptake by bacteria in the presence of various glucose concentrations. The rate of oxygen uptake by the bacteria was expressed as the respiratory quotient. The result indicated that the oxygen uptake was proportional to the glucose concentration. The expecting equation of the regression line was Y=7.7+0.12X where Y: respiratory quotient, ${\mu}l.\;O_2$ taken up/mg. dry wt. bacterium/hr. X: concentration of glucose, mg/l
The aim of the presented paper was to elucidate the physiological background of ethanol inhibition on glucose uptake, ethanol production and cell growth in Z. mobilis. Data obtained from batch and continuous cultures showed that the rates of glucose uptake and ethanol production were not affected but growth rate was apparently reduced by ethanol produced. In order to know the effects of ethanol on the anabolism and the catabolism in Z. mobilis, enzyme activities of the Enter-Doudoroff pathway, viz. hexokinase, glucose 6-phosphate dehydrogenase, were analyzed with the cell grown at different concentration of ethanol produced. As results, it was found that the activities of the glucose kinase and the glucose 6-phosphate dehydrogenase were not affected greatly by the concentration of ethanol where the glucose uptake rates revealed a relatively constant value. However it was very interesting to note that transketolase, which is an essential enzyme to provide the important precursors for cell growth, was affected more apparently to reduce by increasing ethanol levels. Those results might suggest that the apparent reduction of growth rate at ethanol concentration above 20 g/$\ell$ would be caused by the reduction of the transketolase activity, which in turn provide less precursor for the cell growth.
The baculovirus/Spodoptera frugiperda (Sf) cell system has become popular for the production of large amounts of the human erythrocyte glucose transporter, GLUT1, heterologously. However, it was not possible to show that the expressed transporter in insect cells could actually transport glucose. The possible reason for this was that the activity of the endogenous insect glucose transporter was extremely high and so rendered transport activity resulting from the expression of exogenous transporter very difficult to detect. Sf21-AE cells are commonly employed as the host permissive cell line to support the baculovirus AcNPV replication and protein synthesis. The cells grow well on TC-100 medium that contains 0.1 % D-glucose as the major carbon source, strongly suggesting the presence of endogenous glucose transporters. However, unlike the human glucose transporter, very little is known about properties of the endogenous sugar transporter(s) in insect cells. Thus, the uptake of 2-deoxy-D-glucose (2dGlc) by Sf21-AE cells and the inhibition of 2dGlc transport in the insect cells by fructose and cytochalasin B were investigated in the present work. The binding assay of cytochalasin B was also performed, which could be used as a functional assay for the endogenous glucose transporter(s) in the insect cells. Sf21-AE cells were infected with the recombinant virus AcNPV-GT or no virus, at a multiplicity of infection (MOI) of 5. Infected cells were resuspended in PBS plus and minus 300 mM fructose, and plus and minus 20 $\mu$M cytochalasin B for use in transport assays. Uptake was measured at 28$^{\circ}C$ for 1 min, with final concentration of 1 mM deoxy-D-glucose, 2-[1,2-$^3$H]- or glucose, L-[l,$^3$H]-, used at a specific radioactivity of 4 Ci/mol. The results obtained demonstrated that the sugar uptake in uninfected cells was stereospecific, and was strongly inhibited by fructose but only poorly inhibitable by cytochalasin B. It is therefore suggested that the Sf21-AE glucose transporter has very low affinity for cytochalasin B, a potent inhibitor of human erythrocyte glucose transporter.
Kang, Young Eun;Choi, Kyeong-Mi;Park, Eunjin;Jung, Won-Beom;Jeong, Heejin;Yoo, Hwan-Soo
Journal of Life Science
/
v.27
no.3
/
pp.289-294
/
2017
Diabetes mellitus is associated with insulin resistance, which leads to down-regulation of insulin signaling and the decreased glucose uptake. Adipocytes are sensitive to insulin, and closely implicated in insulin resistance and diabetes. Insulin stimulates differentiation of preadipocytes to adipocytes, and increases glucose transport. Allium species have been used as traditional medicine and health-promoting foods. Allium hookeri (A. hookeri) is reported to improve the pancreatic ${\beta}-cell$ damage and exhibit pancreatic anti-inflammatory activity in streptozotocin-induced diabetic rats. We investigated whether A. hookeri extract (AHE) may stimulate glucose uptake in adipocytes through increasing insulin sensitivity. AHE enhanced fat accumulation, a differentiation biomarker, under the partial induction of differentiation by insulin. $PPAR{\gamma}$, a transcription factor highly expressed in adipocytes, promotes adipocyte differentiation and insulin sensitivity. AHE increased the differentiation of preadipocytes through up-regulation of $PPAR{\gamma}$. The activation of $PPAR{\gamma}$ increases the GLUT4 expression during adipocyte differentiation. GLUT4 is responsible for glucose uptake into the adipocytes. AHE increased the expression of GLUT4 in adipocytes, and subsequently enhanced the insulin-stimulated glucose uptake. These results suggest that AHE promotes adipocyte differentiation through activation of $PPAR{\gamma}$, and leads to enhance glucose uptake in adipocytes along with GLUT4 up-regulation. Thus, AHE may be effective for the insulin-sensitizing and anti-diabetic activities.
Objectives : The Herb-combined Remedy(HCR) for diabetes mellitus is known as an anti-hyperglycaemic agent. But its exact mechanisms are unclear. The present study was carried out to investigate its anti-hyperglycaemic and anti-oxidative effects in STZ-induced diabetic rats. Methods : Experimental diabetes was induced by injection of STZ(80mg/kg) to ratsvia the peritoneum. The experimental animals were divided into 4 groups : normal group, control group(STZ-induced diabetic rats with no treatment), HCR group(STZ-induced diabetic rats with HCR treatment), MF group(STZ-induced diabetic rats with Metformin treatment). The effects of HCR on STZ-induced diabetes was observed by measuring fasting blood glucose, changes of body weight, food uptake, and water uptake glucose levels in the normal state decline rates in blood glucose levels DPPH free-radical scavenging activity superoxide dismutase in RBC lysate catalase activity in RBC lysate and glutathione reductase activity in RBC lysate. Results : Treatment with HCR regulated blood glucose levels. Treatment with HCR also prevented weight loss in STZ-induced diabetic rats. In addition, oral glucose tolerance decreased following treatment with HCR. Direct anti-oxidative effects on DPPH free-radical scavenging were not observed, but treatment with HCR elevated SOD levels in blood cell lysates from STZ-induced diabetic rats. In addition, the HCR-treatment group showed an elevated tendency to glutathione reductase activity. Conclusions : These results demonstrate that HCR has anti-hyperglycaemic and anti-oxidative effects in STZ-induced diabetic rats.
A fluorescent glucose analogue,2-[N-(7-nitrobenz-2-ox a-1,3-diazol-4-yl) amino] -2- deoxy-D-glucose (2-NBDG), which had previously been developed for the analysis of glucose uptake in living cells, was investigated to determine its biological activity on microorganisms.2-NBDG did not show any inhibitory effect on growth of yeast cells and bacteria. In contrast, 2-NBDG exhibited strong inhibitory effects on filamentous fungal growth. The growth of filamentous fungi was completely inhibited, when 2-NBDG was supplemented as sole carbon source. The inhibitory effect was decreased by the addition of glucose in the test medium. Furthermore, 2-NBDC inhibited chitinase activity of Trichoderma sp. These results suggested that the inhibitory effects of 2-NBDG on filamentous fungi might be partially due to the inhibition of chitinase.
Lee, Jae Sung;Kang, Yun Hwan;Kim, Kyoung Kon;Yun, Yeong Kyeong;Lim, Jun Gu;Kim, Tae Woo;Kim, Dae Jung;Won, Sang Yeon;Bae, Moo Hoan;Choi, Han Seok;Choe, Myeon
Journal of Nutrition and Health
/
v.47
no.1
/
pp.12-22
/
2014
Purpose: This study was conducted to establish the production conditions through optimization of the production process of beverages using Aspergillus oryzae CF1001, and to analyze volatile compounds and antidiabetic activity. Methods: The optimum condition was selected using the response surface methodology (RSM), through a regression analysis with the following independent variables gelatinization temperature (GT, $X_1$), saccharogenic time (ST, $X_2$), and dependent variable; ${\Delta}E$ value (y). The condition with the lowest ${\Delta}E$ value occurred with combined 45 min ST and $50^{\circ}C$ GT. The volatile compounds were analyzed quantitatively by GC-MS. Results: Assessment of antidiabetic activity of saccharogenic mixed grain beverage (SMGB) was determined by measurement of ${\alpha}$-glucosidase inhibition activity, and glucose uptake activity and glucose metabolic protein expression by reverse transcriptase polymerase chain reaction (RT-PCR) and western blot analysis. Results of volatile compounds analysis, 62 kinds of volatile compounds were detected in SMGB. Palmitic acid (9.534% ratio), benzaldehyde (8.948% ratio), benzyl ethyl ether (8.792% ratio), ethyl alcohol (8.35% ratio), and 2-amyl furan (4.826% ratio) were abundant in SMGB. We confirmed that ${\alpha}$-glucosidase inhibition activity, glucose uptake activity, and glucose-metabolic proteins were upregulated by SMGB treatment with concentration dependent manner. Conclusion: Saccharogenic mixed grain beverage (SMGB) showed potential antidiabetic activity. Further studies will be needed in order to improve the taste and functionality of SMGB.
Objective: Adipose tissue plays a key role in the development of obesity and diabetes. We previously reported that lignosulfonic acid suppresses the rise in blood glucose levels through the inhibition of ${\alpha}$-glucosidase activity and intestinal glucose absorption. The purpose of this study is to examine further biological activities of lignosulfonic acid. Methods: In this study, we examined the effect of lignosulfonic acid on differentiation of 3T3-L1 cells. Results: While lignosulfonic acid inhibited proliferation (mitotic clonal expansion) after induction of differentiation, lignosulfonic acid significantly increased the size of accumulated lipid droplets in the cells. Semi-quantitative reverse transcription polymerase chain reaction analysis showed that lignosulfonic acid increased the expression of the adipogenic transcription factor, peroxisome proliferator-activated receptor gamma ($PPAR{\gamma}$), leading to increased glucose transporter 4 (Glut-4) expression and 2-deoxyglucose uptake in differentiated 3T3-L1 cells. Additionally, feeding lignosulfonic acid to diabetic KK-Ay mice suppressed increase of blood glucose level. Conclusion: Lignosulfonic acid may be useful as a functional anti-diabetic component of food.
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