• Biomolecules & Therapeutics
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• 제16권1호
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• pp.14-20
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• 2008

In the present study, we examined how the transport of choline is regulated at the blood-brain barrier (BBB) under the central nervous system (CNS) cellular damages by oxidative stress using a conditionally immortalized rat brain capillary endothelial cells (TR-BBB), in vitro the BBB model. It was also tested whether the choline uptake is influenced by membrane potential, extracellular pH, protonophore (FCCP) and amiloride in TR-BBB cells. In result, $[^3H]choline$ uptake was inhibited by FCCP and dependent on extracellular pH. The treatment of TR-BBB cells with 20 ng/mL tumor necrosis $factor-{\alpha}$ $(TNF-{\alpha})$, 10 ng/mL lipopolysaccharide (LPS), 100 ${\mu}M$ diethyl maleate (DEM) and 100 ${\mu}M$ glutamate resulted in 3.0-fold, 2.6-fold, 1.8-fold and 2.0-fold increases of $[^3H]choline$ uptake at the respective peak time, respectively. In contrast, hydrogen peroxide and raffinose did not show any significant effects on choline uptake. In addition, choline efflux was significantly inhibited by $TNF-{\alpha}$, LPS and DEM producing cell damage states. In conclusion, the influx and efflux transport system for choline existed in TR-BBB cell line and this process was affected by several oxidative stress inducing agents.

• Parasites, Hosts and Diseases
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• 제61권4호
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• pp.428-438
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• 2023

Clonorchis sinensis is commonly found in East Asian countries. Clonorchiasis is prevalent in these countries and can lead to various clinical symptoms. In this study, we used overlap extension polymerase chain reaction (PCR) and the Xenopus laevis oocyte expression system to isolate a cDNA encoding the choline transporter of C. sinensis (CsChT). We subsequently characterized recombinant CsChT. Expression of CsChT in X. laevis oocytes enabled efficient transport of radiolabeled choline, with no detectable uptake of arginine, α-ketoglutarate, p-aminohippurate, taurocholate, and estrone sulfate. Influx and efflux experiments showed that CsChT-mediated choline uptake was time- and sodium-dependent, with no exchange properties. Concentration-dependent analyses of revealed saturable kinetics consistent with the Michaelis-Menten equation, while nonlinear regression analyses revealed a Km value of 8.3 µM and a Vmax of 61.0 pmol/oocyte/h. These findings contribute to widen our understanding of CsChT transport properties and the cascade of choline metabolisms within C. sinensis.

• 대한핵의학회지
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• 제35권3호
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• pp.185-191
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• 2001

목적: 최근 들어 $[^{11}C]$choline은 양전자방출단층촬영을 통한 각종 종양을 진단하는데 유용한 방사성의약품으로 보고되고 있다. 그러나 C-11의 반감기가 너무 짧은 단점 때문에 일상적인 작업이 쉽지않고, 잦은 합성이 필요하다. $[^{11}C]$Choline의 단점은 반감기가 적당한 F-18이 치환된 $[^{18}F]$fluorocholine을 합성하여 해결할 수 있다. 대상 및 방법: $[^{11}C]$Choline은 $[^{11}C]CH_3I$와- N,N-dimethylaminoethanol을 반응시켜 합성하였고, $[^{18}F]$fluorocholine은 $CH_2Br[^{18}F]$F과 N,N-dimethylaminoethanol을 반응시켜 합성하였다. 방사화학적 순도는 고성능액체크로바토그래프를 이용하여 확인하였다. 두 화합물의 체내동태는 balb/c mouse를 이용하여 5, 20, 40, 80분에 측정하였다. 세포섭취 실험은 glioma (9L)과 colon adenocarcinoma (SW620)를 사용하였다. 결과: 정제후 방사화학적 순도는 98% 이상이었다. 간에서의 섭취는 시간에 따라 변하지 않았고 $[^{11}C]$choline은 약 20 %ID/g, $[^{18}F]$fluorocholine은 약 13 %ID/g 섭취되었다. 신장에서의 섭취는 시간에 따라 감소하였고, 주사 80분 후에 각각 15 %ID/g, 20 %ID/g 섭취되었다. 9L과 SW620세포섭취 실험에 있어서 $[^{11}C]$choline은 각각 4.93, 18.69 %ID/g, $[^{18}F]$fluorocholine은 각각 1.77, 2.77 %ID/g이 섭취된 것을 확인하였다. 결론: $[^{11}C]$Choline과 $[^{18}F]$fluorocholine은 종양 세포주에 따라 다른 섭취 경향을 나타내는 것을 확인하였다. 이상의 결과들로 보아 다른 종양 세포주에 있어서 두 화합물의 섭취 경향에 관한 연구가 필요하다.

• Asian-Australasian Journal of Animal Sciences
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• 제31권5호
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• pp.686-695
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• 2018

Objective: The objective of this study was to investigate the effects of dietary choline supplementation on hepatic lipid metabolism and gene expression in finishing pigs with intrauterine growth retardation (IUGR). Methods: Using a $2{\times}2$ factorial design, eight normal birth weight (NBW) and eight IUGR weaned pigs were fed either a basal diet (NBW pigs fed a basal diet, NC; IUGR pigs fed a basal diet, IC) or a diet supplemented with two times more choline than the basal diet (NBW pigs fed a high-choline diet, NH; IUGR pigs fed a high-choline diet, IH) until 200 d of age. Results: The results showed that the IUGR pigs had reduced body weight compared with the NBW pigs (p<0.05 from birth to d 120; p = 0.07 from d 120 to 200). Increased (p<0.05) free fatty acid (FFA) and triglyceride levels were observed in the IUGR pigs compared with the NBW pigs. Choline supplementation decreased (p<0.05) the levels of FFAs and triglycerides in the serum of the pigs. The activities of malate dehydrogenase and glucose 6-phosphate dehydrogenase were both increased (p<0.05) in the livers of the IUGR pigs. Choline supplementation decreased (p<0.05) malate dehydrogenase activity in the liver of the pigs. Gene expression of fatty acid synthase (FAS) was higher (p<0.05) in the IC group than in the other groups, and choline supplementation decreased (p<0.05) FAS and acetyl-CoA carboxylase ${\alpha}$ expression in the livers of the IUGR pigs. The expression of carnitine palmitoyl transferase 1A (CPT1A) was lower (p<0.05) in the IC group than in the other groups, and choline supplementation increased (p<0.05) the expression of CPT1A in the liver of the IUGR pigs and decreased (p<0.01) the expression of hormone-sensitive lipase in both types of pigs. The gene expression of phosphatidylethanolamine N-methyltransferase (PEMT) was higher (p<0.05) in the IC group than in the other groups, and choline supplementation significantly reduced (p<0.05) PEMT expression in the liver of the IUGR pigs. Conclusion: In conclusion, the lipid metabolism was abnormal in IUGR pigs, but the IUGR pigs consuming twice the normal level of choline had improved circulating lipid parameters, which could be related to the decreased activity of nicotinamide adenine dinucleotide phosphate-generating enzymes or the altered expressions of lipid metabolism-related genes.

• 대한수의학회지
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• 제4권1호
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• pp.15-17
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• 1964

Scabby burley collected in Kyong Sang Nam Do fed to healthy dogs, age less than 2 years, old and determined the Choline-Esterase Activity in blood of dogs. The results obtained in this investigation are summarized as follows. 1. Choline-Esterase Activity in the blood of dogs fed Scabby barley has been decreased. 2. The poisionous component of the Scabby barley thought to be Anticholinesterase.

• Journal of Animal Science and Technology
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• 제58권8호
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• pp.30.1-30.12
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• 2016

Background: The effect of choline chloride supplementation through forced drinking combined with concentrate diets containing Ca-fish oil on milk production and milk composition of Etawah Grade goats was evaluated. Choline chloride is an essential component in ruminant diets as it is required for fat metabolism. Method: The experiment was conducted in a completely randomized block design with three types of treatments and eight replications. The trial had two successive experimental periods; the first, during the eight weeks of late pregnancy, and the second, during the first 12 weeks of lactation. Twenty-four Etawah Grade does in the second gestation period were divided into three treatment groups. Commercial choline chloride 60 % in corncobs-based powder was used as a source of choline chloride. The treatments were no supplementation (control) and supplemented with either 4 g or 8 g/2days of choline chloride. Choline chloride was given to the animals through a forced drinking technique, after dissolving it in 60 ml drinking water. The initial body weight of does was $38.81{\pm}3.66kg$. The does were penned individually, and were given fresh chopped King Grass ad libitum and 700 g/day of concentrate diets containing Ca-fish oil, starting eight weeks prior to expecting kidding and continuing for 12 weeks of parturition. Results: All nutrient intakes were not significantly different (p > 0.05) among the treatments during the late pregnancy and the lactation periods. Supplementation did not affect (p > 0.05) the average daily gains and feed conversion ratio during pregnancy but gave effects (p < 0.05) on the average daily gains, feed conversion ratio and income over feed cost during lactation. The highest average daily milk yields and 4 % fat corrected milk yields were found in goats supplemented with 4 g/2days of choline chloride and increased by 17.00 % and 24.67 %, respectively, compared to the control. Moreover, milk composition percentage and milk constituent yields improved significantly (p < 0.05) in those supplemented with 4 g/2days of choline chloride. Conclusion: The supplementation of 4 g/2days of choline chloride through forced drinking increased milk yields, the 4 % fat corrected milk yields, milk composition, milk constituent yields, and improved feed conversion ratio and income over feed cost of Etawah Grade goats.

• 한국응용과학기술학회지
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• 제3권2호
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• pp.49-64
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• 1986

This experiment is carried out to study effect of choline-deficient diet on serum and liver lipid contents of male rats. The experimental animals use 36 male Sprague-Dawley rats weighing about $100{\pm}3g$. They are classified into 7 groups and fed to experimental diets which are composed of 0.8% choline-supplement of deficient diets in addition to 14% corn oil, 14% corn margarine and 14% lipids mixed with 4% corn oil and 10% corn margarine, respectively. After feeding for 4 weeks, I measure lipid concentration of serum and liver, and the result are as follows. 1. The choline-deficient diet group decreases slightly the rates of weight gain and feed efficiency as compared with those of the choline-supplement diet group, but increases liver weight. 2. The choline-dificient diet group decreases the serum concentrations of total cholesterol (TC), free cholesterol, HDL-choelsterol, VLDL, LDL-cholesterol and phospholipid (PL), but increases the contents of triglyceride and the ratios of cholesterol/HDL-cholesterol and triglyceride/PL, and indicates no remarkable-difference in the ratio of TC/PL. 3. As compared with the choline-supplement diet group, the choline-deficient diet group contains the higher liver contents of total lipids, free cholesterol and triglyceride, and gives little difference in the liver contents of total cholesterol and phospholipid(PL), and presents the higher ratios of VLDL, LDL-cholesterol/HDL-cholesterol and TG/PL in the liver.4. In the choline-deficient diet group, the coutents of serum and liver lipid is not influenced by the kind of dietary fat. On the other hand, the choline-supplemented diet group indicates a significantly lower content of phospholipid in the corn margarine-added diet group than in the corn oil-added diet group. As aforementioned results, I think that the choline-deficient diet induces fatty liver in male rats without relation to kind of fatty acid, and cholines-upplement diet with saturated fatty acid makes the more decrease of liver phospholipid than that with polyunsaturated fatty acid.

• Journal of Nutrition and Health
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• 제41권5호
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• pp.428-438
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• 2008

본 연구는 한국인 상용 식품의 콜린 함량을 분석하기 위하여 2001년도 국민건강영양조사의 다소비 식품과 다빈도 식품 185종에 대하여 choline oxidase 효소법을 사용하여 식품의 총 콜린함량을 분석하여 다음과 같은 결과를 얻었다. 식품 100 g당 총 콜린 함량이 높은 식품은 메추리알 476.04mg, 마른 오징어가 452.42 mg, 소고기간 427.16 mg, 돼지고기간 424.92 mg, 참치통조림 414.44 mg, 멸치 381.30mg, 북어 378.88 mg, 달걀 309.88 mg, 닭고기간 259.38mg, 대두 238.62 mg로 건조어류, 동물의 간, 난류와 두류가 고콜린 식품으로 분류될 수 있겠고, 그 뒤를 이어 마늘빵이 193.18 mg, 쌀보리가 182.73 mg으로 곡류도 상당한 콜린 급원식품인 것으로 나타났다. 채소류와 과일류에서 낮은 콜린 함량을 보였으며, 또한 유자차, 녹차에서도 1 mg 이하의 낮은 콜린 함량을 보였다. 고춧가루, 맥주, 소주, 콩기름, 옥수수 기름 등에서는 콜린이 검출되지 않았다. 채소류, 과일류, 조미료류와 식물성 유지류가 저 콜린식품임을 알 수 있었다. 이상의 결과에서 볼 때 건조어류, 어류 가공품과 간, 난류, 두류, 곡류가 고콜린군 식품이며, 식물성 식품에서는 두류와 곡류 및 그 제품이, 동물성 식품에서는 어패류, 난류, 육류가 콜린의 주요 급원식품임을 알 수 있었다. 앞으로 Zeisel의 결과에 비해 크게 상이한 결과를 보인 쌀이나 밀가 루 등 일부 식품에 대한 보다 정확한 분석절차에 대한 연구가 뒤따라야 하리라고 본다.

• Animal Bioscience
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• 제35권11호
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• pp.1787-1799
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• 2022

Objective: Choline deficiency, one main trigger for nonalcoholic fatty liver disease (NAFLD), is closely related to lipid metabolism disorder. Previous study in a choline-deficient model has largely focused on gene expression rather than gene structure, especially sparse are studies regarding to alternative splicing (AS). In modern life science research, primary hepatocytes culture technology facilitates such studies, which can accurately imitate liver activity in vitro and show unique superiority. Whereas limitations to traditional hepatocytes culture technology exist in terms of efficiency and operability. This study pursued an optimization culture method for duck primary hepatocytes to explore AS in choline-deficient model. Methods: We performed an optimization culture method for duck primary hepatocytes with multi-step digestion procedure from Pekin duck embryos. Subsequently a NAFLD model was constructed with choline-free medium. RNA-seq and further analysis by rMATS were performed to identify AS events alterations in choline-deficency duck primary hepatocytes. Results: The results showed E13 (embryonic day 13) to E15 is suitable to obtain hepatocytes, and the viability reached over 95% by trypan blue exclusion assay. Primary hepatocyte retained their biological function as well identified by Periodic Acid-Schiff staining method and Glucose-6-phosphate dehydrogenase activity assay, respectively. Meanwhile, genes of alb and afp and specific protein of albumin were detected to verify cultured hepatocytes. Immunofluorescence was used to evaluate purity of hepatocytes, presenting up to 90%. On this base, choline-deficient model was constructed and displayed significantly increase of intracellular triglyceride and cholesterol as reported previously. Intriguingly, our data suggested that AS events in choline-deficient model were implicated in pivotal biological processes as an aberrant transcriptional regulator, of which 16 genes were involved in lipid metabolism and highly enriched in glycerophospholipid metabolism. Conclusion: An effective and rapid protocol for obtaining duck primary hepatocytes was established, by which our findings manifested choline deficiency could induce the accumulation of lipid and result in aberrant AS events in hepatocytes, providing a novel insight into various AS in the metabolism role of choline.

• Asian-Australasian Journal of Animal Sciences
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• 제31권11호
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• pp.1795-1806
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• 2018

Objective: The study was designed to establish choline deficiency model (CDM) in broilers for evaluating efficacy of polyherbal formulation (PHF) in comparison with synthetic choline chloride (SCC). Methods: A total of 2,550 one-day-old Cobb 430 broiler chicks were randomly assigned to different groups in three experiments. In experiment 1, G1 and G2 served as normal controls and were fed a basal diet with 100% soybean meal (SBM) as a major protein source supplemented with and without SCC, respectively. In G3, G4, G5, and G6 groups, SBM was replaced at 25%, 50%, 75%, and 100% by soy protein isolate (SPI) to induce a graded level of choline deficiency. In experiment 2, PHF (500 and 1,000 g/ton) in comparison with SCC (1,000 g/ton) were evaluated. In experiment 3, dose-response of PHF (200, 400, and 500 g/ton) with SCC (400 g/ton) was determined. Results: Replacement of SBM by SPI produced a linear decrease in body weight gain (BWG) with a poor feed conversion ratio (FCR). 25% SBM replacement by SPI yielded an optimum negative impact on BWG and FCR; hence, it is considered for further studies. In experiment 2, PHF (500 and 1,000 g/ton) and SCC (1,000 g/ton) showed a similar performance in BWG, FCR and relative liver weight. In experiment 3, PHF produced an optimum efficacy at 400 g/ton and was comparable to SCC in the restoration of serum aspartate aminotransferase activity, abdominal fat, breast muscle lipid content and liver histopathological abnormalities. Conclusion: Replacement of SBM by SPI caused choline deficiency characterised by worsening of BWG, FCR, elevation in liver enzymes and histopathological changes indicating fatty liver. CDM was found valid for evaluating SCC and PHF. It is concluded that PHF has the potential to mimic biological activities of SCC through the restoration of negative effects caused by CDM.