• Archives of Pharmacal Research
• /
• v.28 no.2
• /
• pp.180-185
• /
• 2005

In the process of investigating anti-obesity effect of Platycodi Radix, we found that aqueous extract of Platycodi Radix might inhibit intestinal absorption of dietary fat by inhibiting pancreatic lipase (PL) activity. In order to clarify the anti-obesity mechanism of Platycodi Radix, activity-guided isolation was performed to find active components. The total saponin fraction of Platycodi Radix appeared to have a potent inhibitory activity against the hydrolysis of triolein emulsified with phosphatidycholine by pancreatic lipase in vitro. Based on these results, further purification of active components yielded 10 known triterpenoidal saponins, among these compounds, platycodin A, C, D, and deapioplatycodin D exhibited significant inhibitory effects on PL at the concentration of $500\;{\mu}g/mL$ with 3.3, 5.2, 34.8, and $11.67\%$ pancreatic lipase activity vs control, respectively. Platycodin D was found to inhibit the PL activity in a dose-dependent manner. Therefore, the anti-obesity effect of Platycodi Radix might be due to the inhibition of pancreatic lipase by its saponins.

• Journal of Microbiology and Biotechnology
• /
• v.26 no.6
• /
• pp.1087-1097
• /
• 2016

An intracellular lipase from Anoxybacillus flavithermus HBB 134 was purified to 7.4-fold. The molecular mass of the enzyme was found to be about 64 kDa. The maximum activity of the enzyme was at pH 9.0 and 50℃. The enzyme was stable between pH 6.0 and 11.0 at 25℃, 40℃, and 50℃ for 24 h. The K_m and V_max of the enzyme for pNPL substrate were determined as 0.084 mM and 500 U/mg, respectively. Glycerol, sorbitol, and mannitol enhanced the enzyme thermostability. The enzyme was found to be highly stable against acetone, ethyl acetate, and diethyl ether. The presence of PMSF, NBS, DTT and β-mercaptoethanol inhibited the enzyme activity. Hg²⁺, Fe³⁺, Pb²⁺, Al³⁺, and Zn²⁺ strongly inhibited the enzyme whereas Li⁺, Na⁺, K⁺, and NH₄⁺ slightly activated it. At least 60% of the enzyme activity and stability were retained against sodium deoxycholate, sodium taurocholate, n-octyl-β-D-glucopyranoside, and CHAPS. The presence of 1% Triton X-100 caused about 34% increase in the enzyme activity. The enzyme is thought to be a true lipase since it has preferred the long-chain triacylglycerols. The lipase of HBB 134 cleaved triolein at the 1- or 3-position.

• Applied Chemistry for Engineering
• /
• v.26 no.5
• /
• pp.569-574
• /
• 2015

Arthrospira platensis has high lipid and pigment (such as chlorophyll and carotenoid) contents and thus evaluated as an important resource in functional food production. The cell growth rate and pigment concentration of EM24 increased by approximately 1.2-fold than those of the wild-type strain (WT). Fluorescence intensity levels in EM24, which were quantified with a lipid triolein standard curve, also increased by approximately 1.5-fold than those in WT (62.9 mg/Lvs. 38.9 mg/L). The analysis of fatty acid profiles indicated that the gamma-linoleic acid level in EM24 increased by 1.5-fold than that in WT.

• Journal of Microbiology and Biotechnology
• /
• v.10 no.6
• /
• pp.764-769
• /
• 2000

The lipase gene(lip) from Bacillus stearothermophilus was recombined in vitro by utilizing the DNA shuffling technique. After four rounds of shuffling, transformation, and screening based on the initial rate of clear zone formation on a tricaprylin plate, a clone (M10) was isolated, the cell extract of which showed about 2.8-fold increased lipase activity. The DNA sequence of the mutant lipase gene (m10) showed 3 base changes, resulting in two cryptic mutations and one amino acid substitution: S113($AGC{\rightarrow}AGT$), L252 ($TTG{\rightarrow}TTA$), and G275E ($GGA{\rightarrow}GAA$). SDS-PAGE analysis revealed that the increased enzyme activity observed in M10 was partly caused by high expression of the m10 lipase gene. The amount of the expressed G275E lipase was estimated to comprise as much as 41% of the total soluble proteins of the cell. The maximum velocity ($V_{max}$) of the purified mutant enzyme for the hydrolysis of olive oil was measured to be 3,200 U/mg, which was 10% higher than that of the parental (WT) lipase (2,900 U/mg). Its optimum temperature for the hydrolysis of olive oil was $68^{\circ}C$ and it showed a typical $Ca^{2+}$-dependent thermostability, properties fo which were the same as those of the WT lipase. However, the mutant enzyme exhibited a high enantiospecificity towards (S)-naproxen compared with the WT lipase. In addition, it showed increased hydrolytic activity towards triolein, tricaprin, tricaprylin, and tricaproin.

• Biomedical Science Letters
• /
• v.11 no.1
• /
• pp.31-36
• /
• 2005

To evaluate the magnetic resonance imaging and electron microscopic findings of the hyperacute stage of cerebral fat embolism in cats and the time needed for the development of vasogenic edema. Magnetic resonance imaging was performed at 30 minutes (group 1, n=9) and at 30 minutes and 1, 2, 4, and 6 hours after embolization with triolein (group 2, n= 10). As a control for group 2, the same acquisition was obtained after embolization with polyvinyl alcohol particles (group 3, n=5). Electron microscopic examination was done in all cats. In group 1, the lesions were iso- or slightly hyperintense on T2-weighted (T2W) and diffusion-weighted (DWIs) images, hypointense on the apparent diffusion coefficient (ADC) map image, and markedly enhanced on the gadolinium-enhanced T1-weighted images (Gd-T1WIs). In group 2 at 30 minutes, the lesions were similar to those in group 1. Thereafter, the lesions became more hyperintense on T2WIs and DWIs and more hypoinfense on the ADC map image. In group 3, the lesions showed mild hyperintensity on T2WIs at 6 hours but hypointensity on the ADC map image from 30 minutes, with a tendency toward a greater decrease over time. Electron microscopic findings revealed discontinuity of the capillary endothelial wall, perivascular and interstitial edema, and swelling of glial and neuronal cells in groups 1 and 2. The lesions were hyperintense on T2WIs and DWIs, hypointense on the ADC map image, and enhanced on Gd-T1WIs. On electron microscopy, the lesions showed cytotoxic and vasogenic edema with disruption of the blood-brain barrier.

• Preventive Nutrition and Food Science
• /
• v.16 no.2
• /
• pp.104-109
• /
• 2011

Recently, we have demonstrated that green tea extract (GTE) decreases the intestinal absorption of benzo[a]pyrene (BAP), which is an extremely lipophilic food contaminant. The present study was conducted to examine if an enteral infusion of GTE would influence the biliary secretion of BAP and lipids in rats. Female rats were fed an AIN-93G diet with or without (control) GTE at 5 g/kg diet for 4 week. Following the 4-week dietary treatment, rats with bile duct cannula were infused continuously for 8 hr at 3.0 mL/hr via a duodenal catheter with a lipid emulsion containing $4.0\;{\mu}mol$ BAP labeled with $^{14}C$ ($^{14}C$-BAP), $20.7\;{\mu}mol$ cholesterol, $452\;{\mu}mol$ triolein, and $3.1\;{\mu}mol$ ${\alpha}$-tocopherol, and $396.0\;{\mu}mol$ Na-taurocholate with or without 76.1 mg GTE powder in PBS buffer (pH, 6.4). Bile was collected hourly via bile cannula for an 8 hr period. Our results showed that bile flow did not differ between groups. However, the biliary secretion of $^{14}C$-BAP was significantly enhanced by GTE infusion, compared with those infused with the lipid emulsion alone. However, GTE did not affect the biliary outputs of cholesterol, fat, phospholipid and ${\alpha}$-tocopherol. These findings indicate that GTE has a profound stimulatory effect on the biliary excretion of BAP in rats, without affecting other biliary lipids. The mechanism(s) by which GTE enhances the biliary secretion of BAP remains to be investigated.

• Journal of Biomedical Engineering Research
• /
• v.25 no.4
• /
• pp.243-251
• /
• 2004

This study was to quantitative analysis compare to dynamic characteristic change of the regional cerebral blood volume (rCBV) after development of cerebral fat embolism in cats using perfusion MR Imaging. Forty-four adult rats were used. Triolein (n = 15), oleic acid (n = 9) and linoleic acid (n = 11) were injected into the internal carotid artery using microcatheter through the transfemoral approach. Polyvinyl alcohol (Ivalon) (n = 9) was injected as a control group. Perfusion MR images were obtained at 30 minutes and 2 hours after embolization, based on T2 and diffusion-weighted images. The data was time-to-signal intensity curve and ΔR$_2$＊ curve were obtained continuously with the aid of home-maid image proc in.leased significantly at 2 hours compared with those of 30 minutes (P＜0.005). In conclusion, cerebral blood flow decreased in cerebral fat embolism immediately after embolization and recovered remarkably in time course. It is thought that clinically informations to dynamic characteristic change of the cerebral hemodynamics to the early finding in cerebral infarction by DWI and PWI

• Journal of Korean Neurosurgical Society
• /
• v.47 no.3
• /
• pp.203-209
• /
• 2010

Objective : The purpose of study was to evaluate the feasibility of brain magnetic resonance (MR) images of the rat obtained using a 1.5T MR machine in several blood-brain barrier (BBB) experiments. Methods : Male Sprague-Dawley rats were used. MR images were obtained using a clinical 1.5T MR machine. A microcatheter was introduced via the femoral artery to the carotid artery. Normal saline (group 1, n = 4), clotted autologous blood (group 2, n = 4), triolein emulsion (group 3, n = 4), and oleic acid emulsion (group 4, n = 4) were infused into the carotid artery through a microcatheter. Conventional and diffusion-weighted images, the apparent coefficient map, perfusion-weighted images, and contrast-enhanced MR images were obtained. Brain tissue was obtained and triphenyltetrazolium chloride (TTC) staining was performed in group 2. Fluorescein isothiocyanate (FITC)-labeled dextran images and endothelial barrier antigen (EBA) studies were performed in group 4. Results : The MR images in group 1 were of good quality. The MR images in group 2 revealed typical findings of acute cerebral infarction. Perfusion defects were noted on the perfusion-weighted images. The MR images in group 3 showed vasogenic edema and contrast enhancement, representing vascular damage. The rats in group 4 had vasogenic edema on the MR images and leakage of dextran on the FITC-labeled dextran image, representing increased vascular permeability. The immune reaction was decreased on the EBA study. Conclusion : Clinical 1.5T MR images using a rat depicted many informative results in the present study. These results can be used in further researches of the BBB using combined clinical MR machines and immunohistochemical examinations.

• Biomedical Science Letters
• /
• v.10 no.4
• /
• pp.515-521
• /
• 2004

To measure regional cerebral blood volume (rCBV) with perfusion MR imaging of cerebral fat embolism by neutral fat and free fatty acids in cats. Triolein (group 1, n=15), oleic acid (group 2, n=9) and linoleic acid (group 3, n=11) were infused into unilateral internal carotid artery using microcatheter through the transfemoral approach. PVA particle was used as a non-fat embolic material in a control group (group 4, n=9). Perfusion-weighted MR image was obtained at 30 minutes and 2 hours postembolization, based on T2-and diffusion-weighted images. The data of lesion and contralateral normal area were transferred to personal computer, time-to-signal intensity curve was drawn and trans for used to △R2/sup */ curve in regular order. The process in the personal computer was done by using the author's developmental image processing program and interactive data language (IDL) softwares. Statistical significance was approved by paired t-test and ANOVA. rCBV of the lesion was decreased comparing to the normal area in all groups. The ratios of rCBV were as follows (group No, at 30 minutes, at 2 hours); group 1,32％, 51％; group 2, 30％, 44％; group 3, 39％, 61％; group 4, 21％, 36％. rCBVs of 2 hours was significantly increased compared to those of 30 minutes in all groups (P<0.005). rCBV was decreased at 30 minutes in cerebral fat embolism and recovered a little, but significantly at 2 hours. Perfusion-weighted images was useful method in offering hemodynamic information in cerebral fat embolism.

• Proceedings of the Korean Society of Toxicology Conference
• /
• 2002.05b
• /
• pp.20-45
• /
• 2002

In this study, we demonstrated the in vitro and in vivo formation of carcinogen-lipid adduct and its correlation with DNA or protein adducts. The lipids from serum or hepatocyte membranes of Spragu-Dawley rats. human serum, and standard major lipids were in vitro reacted with benzo[a]pyrene(BP) and BP metabolites. 7,8-Dihydroxy-9,10-epoxy-7,8,9,10-tetrahydrobenzo[a]-pyrene(BPDE-I), an ultimate carcinogenic form of BP, was covalently bound to triglyceride(TG). BPDE-I-TG adducts isolated by thin-layer chromatography (TLC) were further detected by high performance liquid chromatography(HPLC). TGs, including triolein, tripalmitin and tristearin, showed positive reactions with BPDE-I. However, cholesterol, phospholipids(Phosphatidylcholine, phosphatidyl-ethanolamine, phosphatidyl-inositol and sphingomyelin) and nonesterified fatty acids(palmitic acid, oleic acid, linoleic acid and stearic acid) did not react with BPDE-I. In addition, other BP metabolites (BP-phenols and -diols) did not react with TG, which TG appeared to be the most reactive lipid yet studied with respect to its ability to form an adduct with BPDE-I. There was a clear-cut dose-respect to its ability to form an adduct with BPDE-I-lipid adduct in vitro between TG and [1,3-3H]BPDE-I. In an animal study, BPDE-I-TG was also formed in the serum of rats orally treated with BP(25 mg/rat). Also, obvious correlations between [3H]BP related-biomolecule adducts (DNA, protein) or lipid damage and the BPDE-I-TG adduct were obtained in various tissues of mice i.p. treated with [3H]BP. These data suggest that TG can form an adduct with BPDE-I, as do other macromolecules (DNA, RNA, and protein). Therefore, a carcinogen-lipid adduct would be a useful biomarker for chemical carcinogenesis research and cancer risk assessment.