ω3 タカ フホウワ シボウサン ノ イ ネンマク ジョウヒ サイボウ シュウフク ニ アタエル エイキョウ

魚脂に含まれるω3多価不飽和脂肪酸(ω3 polyunsaturated fatty add : ω3 PUFA)であるイコサペント酸エチル(eicosapentaenoic acid : EPA)は潰瘍性大腸炎などに対して臨床効果があると報告されている.一方, EPAは胃潰瘍の予防に対して有効であるというin vivoの報告もある.そこで今回我々は胃上皮細胞の倍養系を用いて上皮細胞の修復に対するEPAの影響を検討した.胃粘膜上皮細胞はラット正常胃粘膜より分離した細胞株であるRGM-1を用いた.Prostaglandin(PG)は細胞上清をELISA法にて測定した.細胞数はWST-8法により測定した.細胞障害は上清のLDHを測定することにより評価した.上皮細胞の修復は,単層培養細胞にスクレーパーを用いて上皮細胞に円形欠損を作成し,時間をおって欠損面積を測定して,速く欠損面積が減少したものを修復促進と評価した.EPAをω6多価不飽和脂肪酸(ω6 polyunsaturated fatty acid : ω6 PUFA)のみを含む培養液に添加すると,上皮細胞修復は濃度依存的に促進した.EPAは,PGE_2の遊離を抑制した.EPAを添加しても生存細胞の数や障害に影響は認められなかった.以上の成績より,ω6 PUFAであるリノール酸のみで,ω3PUFAが存在しない培養液では,上皮修復に不利でありω3PUFAであるEPAを補うことにより修復は回復すると結論された.BACKGROUND Epidemiological studies have shown that a high consumption of fish is associated with a low prevalence of peptic ulcers, which is still contraversial. The beneficial effect of eicosapentaenoic acid (EPA), which is abundant in fish oil, on gastric ulcers and on ulcerative colitis has been climically demonsrated. However, the mechanism of the anti-ulcer action of EPA is not known. In the present study, we investigated the effect of EPA on gastric restitution using in vitro culture restitution model. METHODS 1. Ras normal gastric epithelial cell line, RGM-1 was used. 2. Epithelial cell restitution was assessed by round wound restitution assay 3. PGE2 was measured by enzyme-linked immunosorbent assay (ELISA). 4. Assessment of cell damages were determined by WST-8 assay (modified MTT assay) and LDH release. RESULTS 1. EPA accelerated the restitution of PGM-1.2. EPA inhibited PGE2 release. 3. EPA did not influence the damage of RGM-1 damage, as assessed by WST-8 assay. 4. EPA did not have any effect on LDH release. CONCLUSION Addition of EPA accelerated the restitution of gastric epithelial cells, when n3 polyunsaturated fatty acid is deficient

Contribution of thermal desorption and liquid–liquid extraction for identification and profiling of impurities in methamphetamine by gas chromatography–mass spectrometry

Impurity profiling of methamphetamine (MA) using thermal desorption (TD) and gas chromatography–mass spectrometry (GC–MS) was examined. Using TD/GC–MS, impurities were extracted and separated under various conditions. Optimal chromatograms were obtained when a 20 mg MA sample was extracted at 120 °C for 3 min using a TD instrument, followed by separation of the extracts using a non-polar capillary column coated with (5%phenyl)-methylpolysiloxane. MA samples from nine different batches were analyzed under optimized conditions. Compounds related to the structure of MA, such as benzaldehyde, benzyl alcohol, amphetamine, cis- and trans-1,2-dimethyl-3-phenylaziridine, dimethylamphetamine, and N-acetylephedrine, were detected in the chromatograms without any laborious extraction procedure. Compounds such as ethanol, diethyl ether, and acetic acid, which are considered reagents and solvents for MA synthesis, were also detected in some of the chromatograms. The numbers and intensities of the peaks detected were different among the samples. Impurity profiling of MA using TD was compared with that using liquid–liquid extraction (LLE). Better reproducibility of peak areas was obtained using LLE, whereas higher intensities and numbers of peaks were detected using TD. Solvents were extracted more effectively using TD. The nine batches of MA were classified using both extraction procedures. The nine batches were divided roughly into two groups using data from LLE. Subsequently, the groups were classified in detail using data from TD. TD can be used to provide supplemental information for LLE, and the combination of these extraction methods can be helpful for impurity profiling of MA

Uptake of 3,4-methylenedioxymethamphetamine and its related compounds by a proton-coupled transport system in Caco-2 cells

3,4-Methylenedioxymethamphetamine (MDMA) is an illegal amphetamine-type stimulant (ATS) that is abused orally in the form of tablets for recreational purposes. The aim of this work is to investigate the absorption mechanism of MDMA and other related compounds that often occur together in ATS tablets, and to determine whether such tablet components interact with each other in intestinal absorption. The characteristics of MDMA uptake by the human intestinal epithelial Caco-2 cell line were investigated. The Michaelis constant and the maximal uptake velocity at pH 6.0 were 1.11 mM and 13.79 nmol/min/mg protein, respectively, and the transport was electroneutral. The initial uptake rate was regulated by both intra- and extracellular pH. MDMA permeation from the apical to the basolateral side was inferior to that in the reverse direction, and a decrease in apical pH enhanced MDMA permeation from the basolateral to the apical side. These facts indicate that this transport system may be an antiporter of H+. However, under physiological conditions, the proton gradient cannot drive the MDMA uptake because it is inwardly directed. Large concentration differences of MDMA itself drive this antiporter. Various compounds with similar amine moieties inhibited the uptake, but substrates of organic cation transporters (OCT1-3) and an H+-coupled efflux antiporter, MATE, were not recognized