• Archives of Pharmacal Research
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• 제26권9호
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• pp.768-772
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• 2003

Tween 80 (Polysorbate 80) is a hydrophilic nonionic surfactant commonly used as an ingredient in dosing vehicles for pre-clinical in vivo studies (e.g., pharmacokinetic studies, etc.). Tween 80 increased apical to basolateral permeability of digoxin in Caco-2 cells suggesting that Tween 80 is an in vitro inhibitor of P-gp. The overall objective of the present study was to investigate whether an inhibition of P-gp by Tween 80 can potentially influence in vivo absorption of P-gp substrates by evaluating the effect of Tween 80 on the disposition of digoxin (a model P-gp substrate with minimum metabolism) after oral administration in rats. Rats were dosed orally with digoxin (0.2 mg/kg) formulated in ethanol (40％, v/v) and saline mixture with and without Tween 80 (1 or 10％, v/v). Digoxin oral AUC increased 30 and 61％ when dosed in 1 ％ and 10％ Tween 80, respectively, compared to control (P＜0.05). To further examine whether the increase in digoxin AUC after oral administration of Tween 80 is due, in part, to a systemic inhibition of digoxin excretion in addition to an inhibition of P-gp in the GI tract, a separate group of rats received digoxin intravenously (0.2 mg/kg) and Tween 80 (10％ v/v) orally. No significant changes in digoxin IV AUC was noted when Tween 80 was administered orally. In conclusion, Tween 80 significantly increased digoxin AUC and Cmax after oral administration, and the increased AUC is likely to be due to an inhibition of P-gp in the gut (i.e., improved absorption). Therefore, Tween 80 is likely to improve systemic exposure of P-gp substrates after oral administration. Comparing AUC after oral administration with and without Tween 80 may be a viable strategy in evaluating whether oral absorption of P-gp substrates is potentially limited by P-gp in the gut.

• Asian-Australasian Journal of Animal Sciences
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• 제25권9호
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• pp.1229-1236
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• 2012

Our previous studies showed that kisspeptin-10 (Kp-10) injected in vivo can markedly increase lipid anabolism in liver of quails. In order to investigate the direct effect of Kp-10 on lipid metabolism of hepatocytes in birds, cells were separated from embryos livers and cultured in vitro with 0, 100 and 1,000 nM Kp-10, respectively. The results showed that after 24 h treatment, cells viability was not affected by 100 nM Kp-10, but showed a mild decrease with 1,000 nM Kp-10 compared to the control cells. Based on the results of the cell viability, 100 nM dosage of Kp-10 was selected for the further study and analysis. Compared with control cells, total cholesterol (Tch) contents in 100 nM treated cells were increased by 51.23%, but did not reach statistical significance, while the level of triglyceride (TG), high density of lipoprotein-cholesterol (HDL-C) and low density of lipoprotein-cholesterol (LDL-C) were significantly increased. Real-time PCR results showed that ApoVLDL-II mRNA expression had a tendency to increase, genes including sterol regulatory element-binding protein-1 (SREBP-1), acetyl coenzyme A carboxylase ${\alpha}$ ($ACC{\alpha}$), carnitine palmitoyltransferase 1 (CPT1), 3-hydroxyl-3-methylglutaryl-coenzyme A reductases (HMGCR) and stearyl coenzyme A dehydrogenase-1 (SCD1) mRNA in hepatocytes were significantly down-regulated by 100 nM Kp-10. However, contrary to its gene expression, SREBP-1 protein expression was significantly up-regulated by 100 nM Kp-10. Some of the significant correlations in mRNA expression were found between genes encoding hepatic factors or enzymes involved in lipid metabolism in liver of birds. These results indicate that Kp-10 stimulates lipid synthesis directly in primary cultured hepatocytes of chickens.

• Asian-Australasian Journal of Animal Sciences
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• 제16권8호
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• pp.1131-1136
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• 2003

Forty-eight individually fed Angus steers (body weight $220kg{\pm}9.1$) were utilized to investigate the effects of copper (Cu) source and concentration on lipid metabolism and carcass quality. Steers were stratified by body weight and initial liver Cu concentration and randomly assigned to one of five groups. Groups were then randomly assigned to treatments. Treatments consisted of: 1) control (no supplemental Cu); 2) 10 mg Cu/kg DM from $CuSO_4$; 3) 10 mg Cu/kg DM from a Cu amino acid complex (Availa Cu) 4) 20 mg Cu/kg DM from $CuSO_4$; and 5) 20 mg Cu/kg DM from Availa Cu. Steers were fed a corn-alfalfa-based growing diet for 56 d. Steers were then switched to a high concentrate finishing diet for 145 d. On day 74 of the finishing phase subcutaneous adipose tissue biopsies were obtained from three steers/treatment to determine basal and stimulated lipolytic rates in vitro. Steers were then slaughtered after receiving the finishing diet for 145 d. Control steers tended (p<0.12) to have lower ceruloplasmin (Cp) activity than Cu supplemented steers. Steers receiving 20 mg Cu/kg DM from Availa Cu had higher (p<0.03) Cp activity than steers receiving 20 mg Cu/kg DM from $CuSO_4$. Plasma non-esterified fatty acids were similar across treatments. Steers receiving 10 mg Cu/kg DM from Availa Cu had higher (p<0.02) total plasma cholesterol concentrations relative to steers receiving 10 mg Cu/kg DM from $CuSO_4$. Steers receiving 20 mg Cu/kg DM from Availa Cu had lower (p<0.03) plasma triglyceride concentrations than steers supplemented with 20 mg Cu/kg DM from $CuSO_4$. Fatty acid profile of longissimus muscle was similar across treatments. Backfat depth tended (p<0.18) to be lower in Cu supplemented steers relative to controls. Steers supplemented with 20 mg Cu/kg DM from Availa Cu had heavier (p<0.03) hot carcass weights and a greater (p<0.02) dressing percentage than steers supplemented with 20 mg Cu/kg DM from $CuSO_4$. Furthermore, in vitro basal (p<0.06) and epinephrine stimulated (p<0.04) lipolytic rates of subcutaneous adipose tissue were higher in Cu supplemented steers relative to controls. The results of this study suggest that Cu supplementation has minimal effects on blood and lean tissue lipid profile. However, it appears that Cu may play a role in lipid metabolism in subcutaneous adipose tissue.

• Biomolecules & Therapeutics
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• 제2권3호
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• pp.236-243
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• 1994

A splenocyte culture system supplemented with liver microsomes was developed to detect immunotoxic chemicals which require metabolic activation using cyclophosphamide as a positive standard. When liver microsomes were added to splenocyte cultures isolated from female B6C3Fl mice, the proliferation of splenocytes by lipopolysaccharide (LPS) was increased and the proliferation by concanavalin A (Con A) was decreased. However, when compared with each corresponding control, cyclophophamide was successfully activated to metabolites capable of suppressing Iymphoproliferative responses. This suppression was clearly dependent upon the amounts of microsomes added and/or the concentration of cyclophosphamide exposed. In these cultures, the proliferation of splenocytes was suppressed when the cells were exposed to cyclophosphamide on the day of culture initiation. On the other hand, microsome was responsible for the increase in LPS mitogenicity and NADPH was responsible for the decrease in Con A mitogenicity. Finally, our present culture system was compared with the hepatocyte-splenocyte coculture system which we had developed earlier. We found that the hepatocyte-splenocyte coculture was better able to activate cyclophosphamide to metabolites capable of suppressing the antibody response to sheep erythrocytes. Although our present culture system was relatively poor to activate cyclophosphamide in cultures for antibody response, it will be useful as a simple screening method to detect suppression of certain in vitro immunotoxic parameters like LPS mitogenicity by chemicals which require metabolism.

• Archives of Pharmacal Research
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• 제22권1호
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• pp.30-34
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• 1999

When liriodendrin or syringin was incubated for 24 h with human intestinal bacteria, two metabolites, (+)-syringaresinol$\beta$--D-glucopyranoside and (+)-syringaresionl, from liriodendrin and one metabolite, synapyl alcohol, from syringin were produced. The metabolic time course of liriodendrin was as follows: at early time liriodendrin was converted to (+)-syringaresinol-$\beta$-D-glucopyranoside, and then (+)-syringaresinol. The in vitro cytotoxicities of these metabolites, (+)-syringaresinol and synapyl alcohol, were superior to those of liriodendrin and syringin.

• 농약과학회지
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• 제2권2호
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• pp.29-38
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• 1998

Phenobarbital sodium(PB) 또는 3-methylcholanthrene(3-MC)이 살충제 carbofuran의 쥐에 대한 독성과 이의 독성경감효과를 구명하기 위하여 이들을 단독 또는 조합으로 경구투여한 후 쥐의 생존율을 조사하였고, 쥐에서 carbofuran 의 in vitro 대사에 미치는 영향을 구명하기 위하여 쥐 간의 추출액에 이들을 단독 또는 조합으로 처리한 후 대사산물을 조사하였다. 쥐에 대한 carbofuran의 $LD_{50}$(96hrs)은 6.9 mg/kg이었고, 주 대사산물의 독성은 3-hydroxycarbofuran > 3-ketocarbofuran > 3-hydroxycarbofuran phenol 순으로 높게 나타났으며, 모화합물보다는 그 독성이 매우 낮았다. 쥐의 생존율은 carbofuran 8.4 mg/kg만을 투여했을 때 0%이었으나 carbofuran과 PB 또는 3-MC 20 mg/kg을 각각 조합투여시 $60{\sim}80%$로 높아졌고, 60 mg/kg 투여시에는 100% 생존하여 PB 및 3-MC의 carbofuran에 대한 독성경감 효과가 매우 컸다. 간 추출액에서 in vitro 대사의 대부분은 microsomal fraction에서 이루어지고 있었다. Carbofuran 단독처리시 주 대사산물은 3-hydroxycarbofuran이었으나 carbofuran과 PB 또는 3-MC 조합처리시 3-ketocarbofuran이었다. 또한, 기질 및 처리별 대사산물의 생성율을 조사한 결과 microsomal fraction에 carbofuran 단독 및 PB 또는 3-MC와의 조합처리 모두 co-factor로서 NADP+G-6-P+G-6-P-DG 첨가시(phase I system) 가장 높았고, $105,000{\times}g$ 상징액에서는 carbofuran 단독처리의 경우 co-factor로서 NADPH+ GSH 첨가시(phase II system)에 그리고 PB 또는 3-MC와 조합처리의 경우 co-factor 중 NADPH+FAD 첨가시(phase II system)에 가장 높았다. 대사산물 생성율은 carbofuran 단독처리보다 carbofuran과 PB 또는 3-MC 조합처리에서 $2{\sim}3$배 높았다.

• Toxicological Research
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• 제18권4호
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• pp.331-339
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• 2002

Phenoxy compounds, 2,4-Dichlorophenol acetoxy acid (2,4-D) and 2,4-dichlorophenol (DCP), are widely used as a hormonal herbicide and intermediate for pesticide manufacturing, respectively. In order to assess the potential of these compounds as endocrine disruptors, we studied the androgenicity of them wing in vivo and in vitro androgenicity assay system. Administration of 2,4-D (50 mg/kg/day, p.o.) or DCP (100 mg/kg/day, p.o.) to rats caused an increase in the tissue weight of ventral prostate, Cowpers gland and glands penis. These increase of androgen-dependent tissues were additively potentiated when rats were simultaneously treated with low dose of testosterone (1 g/kg, s.c.). 2,4-D increased about 350% of the luciferase activity in the PC cells transiently cotransfected phAR and pMMTV-Luc at concentration of $10^{-9}$ M. In 2,4-D or DCP-treated castrated rats, testosterone 6$\beta$-hydroxylase activity was not significantly modulated even when rats were co-treated with testosterone. In vitro incubation of 2,4-D and DCP with microsomes at 50 $\mu$M inhibited testosterone 6$\beta$-hydroxylase activity about 27% and 66% in rat liver microsomes, about 44% and 54% in human liver microsomes and about 50% and 45% in recombinant CYP3A4 system, respectively. The amounts of total testosterone metabolites were reduced about 33% and 75% in rat liver microsomes, 69% and 73% in human liver microsomes and 54% and 64% in recombinant CYP3A4 by 2,4-D or DCP, respectively. Therefore, the additive androgenic effect of 2,4-D or DCP by the co-administration of the low dose of testosterone may be due to the increased plasma level of testosterone by inhibiting the cytochrome P450-mediated metabolism of testosterone. These results collectively suggested that 2,4-D and DCP may act as androgenic endocrine disrupter by binding to the androgen receptor as well as by inhibiting the metabolism of testosterone.

• Asian-Australasian Journal of Animal Sciences
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• 제30권6호
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• pp.878-885
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• 2017

Objective: Glucose is an essential fuel in the energy metabolism and synthesis pathways of all mammalian cells. In lactating animals, glucose is the major precursor for lactose and is a substrate for the synthesis of milk proteins and fat in mammary secretory (alveolar) epithelial cells. However, clear utilization of glucose in mammary cells during lactogenesis is still unknown, due to the lack of in vitro analyzing models. Therefore, the objective of this study was to test the reliability of the mammary alveolar (MAC-T) cell as an in vitro study model for glucose metabolism and lactating system. Methods: Undifferentiated MAC-T cells were cultured in three types of Dulbecco's modified Eagle's medium with varying levels of glucose (no-glucose: 0 g/L, low-glucose: 1 g/L, and high-glucose: 4.5 g/L) for 8 d, after which differentiation to casein secretion was induced. Cell proliferation and expression levels of apoptotic genes, Insulin like growth factor-1 (IGF1) receptor, oxytocin receptor, ${\alpha}S1$, ${\alpha}S2$, and ${\beta}$ casein genes were analyzed at 1, 2, 4, and 8 d after differentiation. Results: The proliferation of MAC-T cells with high-glucose treatment was seen to be significantly higher. Expression of apoptotic genes was not affected in any group. However, expression levels of the mammary development related gene (IGF1 receptor) and lactation related gene (oxytocin receptor) were significantly higher in the low-glucose group. Expressions of ${\alpha}S1-casein$, ${\alpha}S2-casein$, and ${\beta}-casein$ were also higher in the low-glucose treated group as compared to that in the no-glucose and high-glucose groups. Conclusion: The results demonstrated that although a high-glucose environment increases cell proliferation in MAC-T cells, a low-glucose treatment to MAC-T cells induces higher expression of casein genes. Our results suggest that the MAC-T cells may be used as an in vitro model to analyze mammary cell development and lactation connected with precise biological effects.

• 대한약학회:학술대회논문집
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• 대한약학회 2001년도 Proceedings of International Convention of the Pharmaceutical Society of Korea
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• pp.306.1-306.1
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• 2001

• 대한핵의학회지
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• 제38권2호
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• pp.198-204
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• 2004

Tumor cell proliferation is considered to be a useful prognostic indicator of tumor aggressiveness and tumor response to therapy but in vitro measurement of individual proliferation is complex and tedious work. PET imaging provides a noninvasive approach to measure tumor growth rate in situ. Early approaches have used $^{18}F$-FDG or methionine to monitor proliferation status. These 2 tracers detect changes in glucose and amino acid metabolism, respectively, and therefore provide only an indirect measure of proliferation status. More recent studies have focused on DNA synthesis itself as a marker of cell proliferation. Cell lines and tissues with a high proliferation rate require high rates of DNA synthesis. $[^{11}C]Thymidine$ was the first radiotracer for noninvasive imaging of tumor proliferation. The short half-life of $^{11}C$ and rapid metabolism of $[^{11}C]Thymidine$ in vivo make the radiotracer less suitable for routing use. Halogenated thymidine analogs such as 5-iodo-2-deoxyuridine (IUdR) can be successfully used as cell proliferation markers for in vitro studies because these compounds are rapidly incorporated into newly synthesized DNA. IUdR has been evaluated as a potential in vivo tracer in nuclear medicing but the image qualify and the calculation of proliferation rates are impaired by its rapid in vivo degradation. Hence, the thymidine analog $3'-deoxy-3'-^{18}F-fluorothymidine$ (FLT) was recently introduced as a stable proliferation marker with a suitable nuclide half-life and stable in vivo. $[^{18}F]FLT$ is phosphorylated to 3-fluorothymidine monophosphate by thymidine kinase 1 and reflects thymidine kinase 1 activity in proliferating cell. $[^{18}F]FLT$ PET is feasible in clincal use and well correlates with cellular proliferation. Choline is a precursor for the biosynthesis of phospholipids (in particular, phosphatidylcholine), which is the essential component of all eukaryotic cell membranes and $[^{11}C]choline$, which is a new marker for cellular proliferation.