• Molecules and Cells
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• v.43 no.1
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• pp.66-75
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• 2020

Saturated fatty acids contribute to β-cell dysfunction in the onset of type 2 diabetes mellitus. Cellular responses to lipotoxicity include oxidative stress, endoplasmic reticulum (ER) stress, and blockage of autophagy. Palmitate induces ER Ca²⁺ depletion followed by notable store-operated Ca²⁺ entry. Subsequent elevation of cytosolic Ca²⁺ can activate undesirable signaling pathways culminating in cell death. Mitochondrial Ca²⁺ uniporter (MCU) is the major route for Ca²⁺ uptake into the matrix and couples metabolism with insulin secretion. However, it has been unclear whether mitochondrial Ca²⁺ uptake plays a protective role or contributes to lipotoxicity. Here, we observed palmitate upregulated MCU protein expression in a mouse clonal β-cell, MIN6, under normal glucose, but not high glucose medium. Palmitate elevated baseline cytosolic Ca²⁺ concentration ([Ca²⁺]_i) and reduced depolarization-triggered Ca²⁺ influx likely due to the inactivation of voltage-gated Ca²⁺ channels (VGCCs). Targeted reduction of MCU expression using RNA interference abolished mitochondrial superoxide production but exacerbated palmitate-induced [Ca²⁺]_i overload. Consequently, MCU knockdown aggravated blockage of autophagic degradation. In contrast, co-treatment with verapamil, a VGCC inhibitor, prevented palmitate-induced basal [Ca²⁺]_i elevation and defective [Ca²⁺]_i transients. Extracellular Ca²⁺ chelation as well as VGCC inhibitors effectively rescued autophagy defects and cytotoxicity. These observations suggest enhanced mitochondrial Ca²⁺ uptake via MCU upregulation is a mechanism by which pancreatic β-cells are able to alleviate cytosolic Ca²⁺ overload and its detrimental consequences.

• Asian-Australasian Journal of Animal Sciences
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• v.13 no.3
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• pp.334-347
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• 2000

The present experiment was carried out to study the utilization of glucose in the mammary gland of crossbred Holstein cattle during feeding with different types of roughage. Sixteen first lactating crossbred Holstein cattle which comprised eight animals of two breed types, Holstein Friesian${\times}$Red Sindhi ($50{\times}50=50%$ HF) and Holstein Friesian${\times}$Red Sindhi ($87.5{\times}12.5=87.5%$ HF). They were divided into four groups of 4 animals each of the same breed. The utilization of glucose in the mammary gland was determined by measuring rates of glucose uptake and the incorporation of glucose into milk components in both groups of 50% HF and 87.5% HF animals feeding on either hay or urea treated rice straw. In early lactation, there were no significant differences of the total glucose entry rate and glucose carbon recycling among groups of crossbred animals feeding on either hay or urea treated rice straw. During lactation advance, the total glucose turnover rates and recycling of carbon glucose of crossbred HF animals feeding on urea treated rice straw were markedly higher than those of crossbred HF animals feeding on hay as roughage, whereas there were no significant changes for both groups of crossbred animals feeding on hay. The percentages and values of non-mammary glucose utilization showed an increase during lactation advance in the same group of both 50% HF and 87.5% HF animals. The percentage of glucose uptake for utilization in the synthesis of milk lactose by the mammary gland was approximately 62% for both groups of 87.5% HF and by approximately 55% for both groups of 50% HF animals feeding on either hay or urea treated rice straw. Intracellular glucose 6-phosphate metabolized via the pentose phosphate pathway accounted for the NADPH (reducing equivalent) of fatty acid synthesis in the mammary gland being higher in 87.5% HF animals during mid-lactation. A large proportion of metabolism of glucose via the Embden-Meyerhof pathway in the mammary gland was more apparent in both groups of 50% HF animals than those of 87.5 % HF animals during early and mid-lactation while it markedly increased for both groups of 87.5% HF animals during late lactation. It can be concluded that utilization of glucose in the mammary gland occurs in a different manner for 50% HF and 87.5% HF animals feeding on either hay or urea treated rice straw. The glucose utilization for biosynthetic pathways in the mammary gland of 50% HF animals is maintained in a similar pattern throughout the periods of lactation. A poorer lactation persistency in both groups of 87.5% HF animals occurs during lactation advance, which is related to a decrease in the lactose biosynthetic pathway.

• Journal of Life Science
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• v.24 no.12
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• pp.1284-1290
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• 2014

Fatty acid transporter protein 1 (FATP1) is highly expressed in skeletal muscle and modulates fatty acid uptake and metabolism. However, the influence of insulin-like growth factor-I (IGF-I), a master regulator of skeletal muscle cells, on FATP1 in skeletal muscle cells has not been demonstrated. To investigate the effect of IGF-I on FATP1 and the expression of the IGFBP5 protein, differentiated C2C12 murine skeletal muscle cells were treated with 20 ng/ml of IGF-I at different time points. The results showed that IGF-I increased FATP1 and IGFBP5 protein expression in a time-dependent manner. To determine whether this induction of FATP1 by the IGF-I treatment was regulated pretranslationally, the mRNA level of FATP1 was measured by real-time quantitative PCR. The IGF-I treatment resulted in very rapid induction of the FATP1 mRNA transcript in C2C12 myotubes. FATP1 mRNA increased 169% and 132% after 24 and 48 h of the IGF-I treatment, respectively, and it returned to control levels after 72 h of the treatment, suggesting that the FATP1 gene is regulated pretranslationally by IGF-I in skeletal muscle cells. This is the first evidence that IGF-I can regulate the expression of FATP1. In conclusion, IGF-I induced rapid transcriptional modification of the FATP1 gene in C2C12 skeletal muscle cells and had modulating effects on fatty acid uptake proteins and oxidative proteins.

• The Korean Journal of Nuclear Medicine
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• v.24 no.2
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• pp.222-228
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• 1990

The development of histomorphometric and histodynamic investigations has permitted the description of a specific and complex osteopathy in hyperthyroidism. The increased bone turnover rate in hyperthyroid patients may be accompanied by a considerable bone loss. These features are associated with both inclosed osteoclastic bone resorption and increased osteoblastric bone formation, with an accelerated calcification rate. Conventional biochemical markers of bone metabolism, i.e. serum calcium and alkaline phosphatase and urinary hydroxyproline and calcium are normal in most patients with hyperthyroidism. However, the correlation between serum BGP and serum concentration of thyroid hormon suggests that serum BGP may be a sensitive marker of increased bone formation due to the hypersecretion of thyroid hormones. Any increase in bone turnover, whether focal or diffuse, will result in an increase in $^{99m}Tc-methylenediphosphonate$ uptake (MDP). The measurement of this uptake in hyperthyroid patients by bone provides a sensitive and objective means of quantifying skeletal metabolism. Using a standard shadow-shield whole-body monitor and radioimmunoassay kit, we have measured whole-body retention of $^{99m}Tc-MDP$ up to 24hr and concentration of serum Osteocalcin in 20 patients with hyperthyroidism and in 42 normals. The results were as follows; 1) The average of serum Osteocalcin level in 42 patients with normals was $9.90{\pm}4.87(ng/ml)$ and in 20 patients with hyperthyroidism was $19.54{\pm}5.7(ng/ml)$. Both the averages of serum Osteocalcin and 24hr $^{99m}Tc-MDP$ uptakes in hyperthyroid patients were higher than those in normals. 2) $^{99m}Tc-MDP$ uptakes in skeletal system increased in proportion to normal ageing after 40 yrs old in 42 patients with normals. The average of $^{99m}Tc-MDP$ uptakes in hyperthyroid patients were higher than those in normals without related ageing. 3) A significant relationships between the $^{99m}Tc-MDP$ uptakes and serum Osteocalcin level were peformed (r=0.55, $y=17.58+6.7\times$). From the above results we concluded that the measurement of serum Osteocalcin and 24hr $^{99m}Tc-MDP$ uptakes can be used for evaluation of bone turnover as a specific marker in hyperthyroid patients.

• The Journal of Internal Korean Medicine
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• v.21 no.3
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• pp.377-388
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• 2000

Objective : This study was undertaken to evaluate the effect of Sunghyangchungisan (SHCS) on the regulation of vascular tone and $Ca^{2+}$ metabolism in arterial tissues. Vascular rings isolated from rabbit carotid artery were myographed isometrically in isolated organ baths and the effect of SHCS on contractile activities, endothelial function and $Ca^{2+}$ metabolism were determined. Methods : In phentobarbital sodium-anesthetized rabbits, SHCS administered through ear vein (100 mg/Kg body wt.) or intragastric dwelling tube (300 mg/Kg body wt.) attenuated phenylephrine (PE, 10 ${\mu}g$/Kg, i.v.)-induced increases in both systolic and diastolic cartoid arterial blood pressure. Results : In experiments with isolated arterial strips, SHCS relaxed arterial rings which were pre-contracted by phenylephrine (PE, 1 ${\mu}M$). The responses to SHCS were partially dose-dependent at concentrations lower than 0.5 mg/ml. When SHCS was applied prior to the exposure to PE, it inhibited the PE-induced contraction by a similar magnitude which was comparable to the relaxation of pre-contracted arterial rings. Washout of SHCS after observing its relaxant effect resulted in a full recovery of PE-induced contractions, indicating that the action mechanism is reversible. The observation that SHCS did not change the $ED_{50)$ of PE oh its dose-response curve ruled out the possible interaction of SHCS with ${\alpha}$-receptors. The relaxant effect of SHCS was not affected by removal of endothelium or a nitric oxide synthase inhibitor, L-NAME. Methylene blue, an inhibitor of the soluble guanylate cyclase, did not affect the relaxant effect of SHCS. These results suggest that the action of SHCS is not mediated by the endothelium nor soluble guanylate cyclase. Constant cGMP production determined in arterial strips in the presence or absence of SHCS is consistent with this conclusion. When contraction was induced by additive application of $Ca^{2+}$ in arterial rings which were pre-depolarized by high $K^+$ in a $Ca^{2+}$-free solution, the relaxant effect of SHCS was attenuated by increasing the $Ca^{2+}$ concentration. SHCS, when applied to the arterial rings pre-contracted by PE and then relaxed by nifedipine, a $Ca^{2+}$ channel blocker, did not show additive relaxation. SHCS partially blocked $Ca^{2+}$ influx stimulated by PE and high $K^+$ which was determined by 5-min ^{45}Ca$ uptake, while it did not affect $Ca^{2+}$ efflux. Conclusions : From above results, it is suggested that SHCS relax PE-induced contraction of rabbit carotid artery in an endothelium independent manner, andinhibition of $Ca^{2+}$ influx may contribute to the underling mechanism.

• Korean Journal of Weed Science
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• v.11 no.3
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• pp.187-194
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• 1991

Absorption and translocation, and metabolism studies using $^{14}C$-naproanilide were conducted to determine selective mode of action of naproanilide in rice and paddy weeds under different temperature conditions. Absorption amount of $^{14}C$-naproanilide was greater in Cyperus serotinus and Sagittaria pygmaea than rice and Echinochloa crusgalli. Especially, absorption of $^{14}C$-naproanilide in C. serotinus was increased twice at 32 $^{\circ}C$ and 48 hour exposure conditions. $^{14}C$-naproanilide in roots was translocated to shoots very little in rice and E. crusgalli, but S. pygmaea somewhat greater translocation than the other species. In C. serotinus and S. pygmaea, susceptible weeds, metabolic rates of naproanilide into phytotoxic NOP (2-(2-naphthoxy)-propionic acid) and NOPM (methyl 2-(2-naphthoxy) propionate) were significantly greater than in rice and E. crugalli, tolerant species. Consequently, differential uptake by roots and the difference in activation metabolism of naproanilide among species may explain the possible mechanism of selectivity.

• BMB Reports
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• v.30 no.1
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• pp.1-6
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• 1997

The ability of Hep-G2 cells to process $[^{125}I]LDL$ under basal conditions was investigated. The receptor-binding and internalization of $[^{125}I]LDL$ increased with the time of incubation in a saturable manner. After 4 h of incubation, 31.4 ng of $[^{125}I]LDL$ was cell bound. The cells rapidly internalized $[^{125}I]LDL$ via specific, receptor-mediated endocytosis. The amount of internalized $[^{125}I]LDL$ reached a maximun of 96.7 ng at 2 h of incubation and remained constant for the next 2 h. The rate of degradation of internalized $[^{125}I]LDL$ proceeded in a linear manner over the entire 4 h of incubation after an initial lag period. The effects of individial fatty acids (C18:0. C18:1, C18:2. and C18:3), differing in their degree of unsaturation. on the receptor-binding, internalization and degradation of $[^{125}I]LDL$ were also investigated. Inclusion of 1.0 mM of each fatty acid into the culture medium significantly increased $[^{125}I]LDL$ metabolism in Hep-G2 cells. Among the fatty acids tested, stearic acid had the least effect on the receptor-binding activity. There were no significant differences among the unsaturated fatty acids in LDL-receptor binding. The effect of individual fatty acids on the $[^{125}I]LDL$ uptake was similar to that of the receptor-binding. showing a significantly lower effect with stearic acid. The amount of degraded material of internalized $[^{125}I]LDL$ was the lowest with stearic acid when it was compared with unsaturated fatty acids.

• Nutrition Research and Practice
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• v.14 no.3
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• pp.203-217
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• 2020

BACKGROUND/OBJECTIVE: Centella asiatica, also known as Gotu kola, is a tropical medicinal plant native to Madagascar, Southeast Asia, and South Africa. It is well known to have biological activities, including wound healing, anti-inflammatory, antidiabetic, cytotoxic, and antioxidant effects. The purpose of this study was to determine the efficacy of extracts of C. asiatica against age-related eye degeneration and to examine their physiological activities. MATERIALS/METHODS: To determine the effects of CA-HE50 (C. asiatica 50% EtOH extract) on retinal pigment cells, we assessed the cytotoxicity of CoCl₂ and oxidized-A2E in ARPE-19 cells and observed the protective effects of CA-HE50 against N-methyl-N-nitrosourea (MNU)-induced retinal damage in C57BL/6 mice. In particular, we measured factors related to apoptosis and anti-oxidation and the protein levels of rhodopsin/opsin. We also measured glucose uptake to characterize glucose metabolism, a major factor in cell protection. RESULTS: Induction of cytotoxicity with CoCl₂ and oxidized-A2E inhibited decreases in the viability of ARPE-19 cells when CA-HE50 was administered, and promoted glucose uptake under normal conditions (P < 0.05). In addition, CA-HE50 inhibited degeneration/apoptosis of the retina in the context of MNU-induced toxicity (P < 0.05). In particular, CA-HE50 at 200 mg/kg inhibited the cleavage of pro-caspase-3 and pro-poly (ADP-ribose)-polymerase and maintained the expressions of nuclear factor erythroid 2-related factor 2 and heme oxygenase-1 similar to normal control levels. Rhodopsin/opsin expression was maintained at a higher level than in normal controls. CONCLUSION: A series of experiments confirmed that CA-HE50 was effective for inhibiting or preventing age-related eye damage/degeneration. Based on these results, we believe it is worthwhile to develop drugs or functional foods related to age-related eye degeneration using CA-HE50.

• The Journal of Internal Korean Medicine
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• v.28 no.1
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• pp.124-132
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• 2007

Objective : This study was to elucidate the effects of Hyunsamkabhangbang (HSK) on the hyperthyroidism induced by sodium levothyroxine. Method : Sprague-Dawley rats were separated into five groups. Except the normal group, the other four groups were treated with sodium levothyroxine 160${\mu}g/kg/day$ for 5 days by oral administration. Among the four groups, all except one (as control) were treated with various concentrations of HSK (0.5, 1.0, 2.0 g/kg/days) for 3 days separately. T3-uptake, T3, T4, TSH, total cholesterol, free fatty acid, ${\beta}$-lipoprotein, glucose, ALP, AST, ALT change in the serum and body weight of rats were measured after medication of solid extract of HSK. Results : HSK decreased significantly serum T3, T4, T3-uptake, ALP, free fatty acid and glucose level in a dose-dependent manner compared with control. HSK increased significantly serum TSH and total cholesterol level and body weight in a dose-dependent manner compared with control. HSK also increased serum ${\beta}$-lipoprotein level compared with control, but not statistically meaningfully. HSK increased significantly serum AST and ALT level compared with control, but relationship to hyperthyroidism is not found. Conclusions : From these results, it could be concluded that HSK lowers excess thyroid hormone and increased metabolism, resulting in improvement of hyperthyroid state.

• Journal of Gastric Cancer
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• v.17 no.4
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• pp.384-393
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• 2017

Purpose: The tumor microenvironment is known to be associated with the metabolic activity of cancer cells and local immune reactions. We hypothesized that glucose metabolism measured by 2-deoxy-2-($^{18}F$)fluoro-D-glucose ($^{18}F-FDG$) positron emission tomography (PET)-computed tomography (CT) ($^{18}F-FDG$ PET-CT) would be associated with local immune responses evaluated according to the presence of tumor infiltrating lymphocytes (TILs). Materials and Methods: We retrospectively reviewed 56 patients who underwent $^{18}F-FDG$ PET-CT prior to gastrectomy. In resected tumor specimens, TIL subsets, including cluster of differentiation (CD) 3, CD4, CD8, Forkhead box P3 (Foxp3), and granzyme B, were subjected to immunohistochemical analysis. The prognostic nutritional index (PNI) was calculated as: ($10{\times}serum$ albumin value)+($0.005{\times}peripheral$ lymphocyte counts). Additionally, the maximum standard uptake value ($SUV_{max}$) was calculated to evaluate the metabolic activity of cancer cells. Results: The $SUV_{max}$ was positively correlated with larger tumor size (R=0.293; P=0.029) and negatively correlated with PNI (R=-0.407; P=0.002). A higher $SUV_{max}$ showed a marginal association with higher CD3 (+) T lymphocyte counts (R=0.227; P=0.092) and a significant association with higher Foxp3 (+) T lymphocyte counts (R=0.431; P=0.009). No other clinicopathological characteristics were associated with $SUV_{max}$ or TILs. Survival analysis, however, indicated that neither $SUV_{max}$ nor Foxp3 held prognostic significance. Conclusions: FDG uptake on PET-CT could be associated with TILs, especially regulatory T cells, in gastric cancer. This finding may suggest that PET-CT could be of use as a non-invasive tool for monitoring the tumor microenvironment in patients with gastric cancer.