• The Korean Journal of Physiology and Pharmacology
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• v.5 no.4
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• pp.287-297
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• 2001

Contraction of smooth muscle is initiated by an increase in cytosolic $Ca^{2+}$ leading to activation of $Ca^{2+}$/ calmodulin-dependnet myosin light chain (MLC) kinase and phosphorylation of MLC. The types of contraction and signaling mechanisms mediating contraction differ depending on the region. The involvement of these different mechanisms varies depending on the source of $Ca^{2+}$ and the kinetic of $Ca^{2+}$ mobilization. $Ca^{2+}$ mobilizing agonists stimulate different phospholipases $(PLC-{\beta},\;PLD\;and\;PLA_2)$ to generate one or more $Ca^{2+}$ mobilizing messengers $(IP_3\;and\;AA),$ and diacylglycerol (DAG), an activator of protein kinase C (PKC). The relative contributions of $PLC-{\beta},\;PLA_2$ and PLD to generate second messengers vary greatly between cells and types of contraction. In smooth muscle cell derived form the circular muscle layer of the intestine, preferential hydrolysis of $PIP_2$ and generation of $IP_3$ and $IP_3-dependent\;Ca^{2+}$ release initiate the contraction. In smooth muscle cells derived from longitudinal muscle layer of the intestine, preferential hydrolysis of PC by PLA2, generation of AA and AA-mediated $Ca^{2+}$ influx, cADP ribose formation and $Ca^{2+}-induced\;Ca^{2+}$ release initiate the contraction. Sustained contraction, however, in both cell types is mediated by $Ca^{2+}-independent$ mechanism involving activation of $PKC-{\varepsilon}$ by DAG derived form PLD. A functional linkage between $G_{13},$ RhoA, ROCK, $PKC-{\varepsilon},$ CPI-17 and MLC phosphorylation in sustained contraction has been implicated. Contraction of normal esophageal circular muscle (ESO) in response to acetylcholine (ACh) is linked to $M_2$ muscarinic receptors activating at least three intracellular phospholipases, i.e. phosphatidylcholine-specific phospholipase C (PC-PLC), phospholipase D (PLD) and the high molecular weight (85 kDa) cytosolic phospholipase $A_2\;(cPLA_2)$ to induce phosphatidylcholine (PC) metabolism, production of diacylglycerol (DAG) and arachidonic acid (AA), resulting in activation of a protein kinase C (PKC)-dependent pathway. In contrast, lower esophageal sphincter (LES) contraction induced by maximally effective doses of ACh is mediated by muscarinic $M_3$ receptors, linked to pertussis toxin-insensitive GTP-binding proteins of the $G_{q/11}$ type. They activate phospholipase C, which hydrolyzes phosphatidylinositol bisphosphate $(PIP_2),$ producing inositol 1, 4, 5-trisphosphate $(IP_3)$ and DAG. $IP_3$ causes release of intracellular $Ca^{2+}$ and formation of a $Ca^{2+}$-calmodulin complex, resulting in activation of myosin light chain kinase and contraction through a calmodulin-dependent pathway.

• Journal of Nutrition and Health
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• v.29 no.1
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• pp.112-121
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• 1996

The study was designed to observe the effect of blend fat calculated from the foods consumed in Korean with those of perilla oil, beef tallow and corn oil on colonic mucosal phospholipid fatty acid composition and the levels of TXB2 and diacylglycerol (DAG) which were known as biomarkers for cancer. Male Sprague Dawley rats, at 7 weeks of age, were divided into control and 1, 2-dimethylhydrazine (DMH)-treated group, and each group was subdivided into four groups. The experimental diets contained one of four dietary fats, blend fat (BF), perilla oil(PO), beef tallow (BT) or corn oil (CO), at 15% (w/w) level. At the same time, each rat was injected with saline for control group or DMH twice a week for 6 weeks to give total dose of 180mg/kg body weight. DMH injection, regardless of the type of dietary fats, significantly increased the levels of PGE2 and TXB2 in colonic mucosal layer compared to control (p<0.01). However, the level of eicosanoids was influenced by the types of dietary fats in both control and DMH group. In control groups, colonic mucosal level of TXB2 was higher in beef tallow group, but lower in perilla oil group compared to that of blend fat (p<0.01). In DMH groups, the level of TXB2 was higher in beef tallow and corn oil groups(p<0.05). The level of PGE2 showed the same trends with TXB2 and beef tallow most significantly increased the level of PGE2. DMH treatment did not influence on tissue fatty acid profile, which was directly reflected by dietary fatty acid composition. Proportions of C18 : 2 in colonic mucosal phospholipid well reflected dietary level of C18 : 2 showing the order CO>BF>PO>BT. The precentage of arachidonic acid(AA) in mucosal phospholipid was the highest by CO adn BT groups and the lowest by PO group. The incorporation of $\alpha$-linolenic acid in colonic mucosal phospholipid in perilla oil group was negatively correlated to the content of AA. Dietary level of C18 : 2 might not be the only controlling factor for the production of eicosanoids in colonic mucosa layer and might function with $\omega$3 fatty acids. The level of DAG was significanlty lower in PO group than that of BT group. Therefore, $\omega$3 $\alpha$-linolenic acid rich perilla oil could be very important dietary sourec in controlling eicosanoid production DAG level in cloln and recommenced to use more often in meal preparation to reduce the risk factor against colon cancer.

• Journal of the Korean Society of Food Science and Nutrition
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• v.38 no.8
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• pp.1062-1068
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• 2009

Diacylglycerol (DAG) were synthesized by enzymatic esterification of glyceryl monooleate (GMO) and conjugated linoleic acid (CLA) in a shaking water bath. The reaction was catalyzed by Lipozyme TLIM (immobilized lipase from Thermomyces lanuginosa). Effects of reaction time, molar ratio, enzyme road and molecular sieves were studied. Results of normal-phase high performance liquid chromatography (NP-HPLC) analysis were performed. At 1:1, 2:1 and 3:1 (GMO : CLA) molar ratio and Lipozyme TLIM of 20% amount, DAG were produced in 42.6, 54.4 and 54.6 area% in 1 hr, respectively. When different Lipozyme TLIM amounts (2, 5, 10, 20%) were used with 2:1 (GMO : CLA) molar ratio, DAG were produced 21.4 (24 hr), 51.7 (12 hr), 56.2 (6 hr) and 54.4 (1 hr) area%, respectively. The reaction in the absence of molecular sieves increased DAG contents. The maximum DAG concentration conditions were obtained with molar ratio of 2:1 (GMO : CLA), lipase concentration of 10% (of substrate), 10% molecular sieves and reaction time of 6 hours at 55$^{\circ}C$. Under this reaction condition, produced DAG-rich oil was composed of 69 area% DAG, 7.9 area% TAG, 2 area% FFA, and 21.1 area% MAG.

• Applied Biological Chemistry
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• v.41 no.6
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• pp.414-420
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• 1998

We studied the difference effects of dietary ${\alpha}-linolenic\;acid\;({\alpha}-LA,\;18:3\;n-3)$, eicosapentaenoic acid (EPA, 20:5 n-3) and docosahexaenoic acid (DHA, 22:6 n-3) on the lowering of triacylglycerol in the liver and serum on lipid metabolism in rats. Rats were fed semipurified diets containing 10% fat with constant polyunsaturated/monounsaturated/saturated fatty acids (1:1:1) and n-6/n-3 ratio (1:2). EPA (98%) and DHA (98%) were added in diets as the ethyl esters. The concentration of liver triacylglycerol was significantly lower in rats fed both EPA and DHA than in those fed ${\alpha}-LA$. The concentration of liver phospholipid was significantly higher in rats fed DHA than in those fed ${\alpha}-LA$ and EPA. Both EPA and DHA reduced serum triacylglycerol concentration compared with ${\alpha}-LA$, but this effect was more pronounced in the EPA diet. The activity of phophatidate phosphohydrolase in the liver microsome was significantly lower in rats fed both EPA and DHA than in those fed ${\alpha}-LA$. but, there was no significant difference on the activities of diacylglycerol acyltransferase among the three groups. The concentration of liver triacylglycerol were correlated with changes in the microsomal phosphatidate phosphohydrolase activity (r=0.84). Hepatic NADPH generating enzyme, glucose-6-phosphate dehydrogenase was more effective to reduce the activity in rats fed both EPA and DHA than in those fed ${\alpha}-LA$. In conclusion, EPA or DHA reduced the hepatic triacylglycerol concentration by inhibiting microsomal phosphatidate phosphohydrolase, thereby inhibiting synthesis of triacylglycerol in the liver.

• Journal of Life Science
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• v.20 no.10
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• pp.1451-1457
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• 2010

Serotonin (5-hydroxytryptamine, 5-HT) is an important mediator of cell-cell signaling in neuronal systems. The serotonin transporter (SERT) on the plasma membrane controls the extracellular 5-HT level by reuptake of released 5-HT from the synaptic cleft, but the underlying regulation mechanism is unclear. Here, we used the yeast two-hybrid system to identify the specific binding protein(s) that interacts with the carboxyl (C)-terminal region of SERT and found a specific interaction with protein kinase C-$\varepsilon$ (PKC-$\varepsilon$), a PKC isotype that is characterized as a calcium-independent and phorbol ester/diacylglycerol-sensitive serine/threonine kinase. PKC-$\varepsilon$ bound to the tail region of SERT but not to other members of the $Na^+/Cl^-$ dependent SLC6 gene family in the yeast two-hybrid assay. The C-terminal region of PKC-$\varepsilon$ is essential for interaction with SERT. In addition, these proteins showed specific interactions in the glutathione S-transferase (GST) pull-down assay. PKC-$\varepsilon$ phosphorylated the peptide of the SERT amino (N)-terminus in vitro. These results suggest that the phosphorylation of SERT by PKC-$\varepsilon$ may regulate SERT activity in plasma membrane.

• Nutritional Sciences
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• v.7 no.4
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• pp.208-213
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• 2004

In this study, we examined the effects of dietary 1,3-diacylglycerol (DG) compared to conventional triacylglycerol (TG) oil on the postprandial response of total and chylomicron TG, glucose, insulin, and free fatty acid (FFA). This study was conducted using a cross-over design. Ninety subjects participated in the high-fat meal tolerance test where they were randomly assigned to consume two experimental sandwiches containing mayonnaise with TG or DG oil with a seven-day interval. Blood samples were collected before ingestion and at 2, 3, 4 and 6 hr time point after ingestion and analyzed for total and chylomicron TG, glucose, insulin, FFA and phospholipid fatty acid composition. Both TG and DG ingestion had similar effects on postprandial TG response, but a different response from chylomicron TG. Compared with the TG group, TG levels were significantly lower only at 6 hr time point in the DG group. On the other hand, chylomicron TG rose steeply at 2 hr time point and decreased faster in this group. Also, the adjusted value to fasting levels was the same as the unadjusted level. Fasting levels and net differences in insulin were significantly lower at 3 hr time point where chylomicron TG levels were significantly lower in the DG group. But those of glucose and FFA in the TG and DG groups did not differ significantly. Fasting and postprandial levels of fatty acid composition in serum phospholipids in the two groups did not differ significantly. In conclusion, this study indicated that one could reduce the magnitude of postprandial lipemia without influencing glucose metabolism by consumning DG oil as a substitute for TG oil. Based on the correlation of coronary artery disease and postprandial lipemia, dietary DG ingestion might have a beneficial effect in treating such a disease. Further studies are required to clarify the long-tenn effects of dietary DG on blood lipid levels in humans.

• The Korean Journal of Physiology and Pharmacology
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• v.1 no.4
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• pp.445-456
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• 1997

We previously reported that some components of ginsenosides decreased mediator releases evoked by the activation of mast cells with specific antigen-antibody reactions. This study aimed to assess the effects of ginsenosides ($Rb_2$, Re) on the mechanism of histamine release in the mast cell activation. We partially purified guinea pig lung mast cells by using enzyme digestion, the rough and the discontinuous percoll density gradient method. Mast cells were sensitized with $IgG_1$ and challenged with ovalbumin (OA). Histamine was assayed by fluorometric analyzer, leukotrienes by radioimmunoassay. Phospholipase D (PLD) activity was assessed more directly by the production of $[^3H]phosphatidylbutanol$ (PBut) which was produced by PLD-mediated transphosphatidylation in the presence of butanol. The amount of 1,2- diacylglycerol (DAG) were measured by the $[^3H]DAG$ labeled with $[^3H]palmitic$ acid or $[^3H]myristic$ acid. Pretreatment of $Rb_2$ ($300\;{\mu}g$) significantly decreased histamine release by 60%, but Re ($300\;{\mu}g$) increased histamine release by 34%. Leukotrienes release in $Rb_2$ was decreased by 40%, Re was not affected in the leukotrienes release during mast cell activations. An increasing PLD activity during mast cell activation was decreased by the dose-dependent manner in the pretreatment of $Rb_2$, but Re pretreatment facilitated the increased PLD activity during mast cell activation. The amount of DAG produced by phospholipase C (PLC) activity was decreased by $Rb_2$ pretreatment, but Re pretreatment was not affected. The amount of mass DAG was decreased by $Rb_2$ and Re pretreatment during mast cell activation. The data suggest that $Rb_2$ purified from Korean Red Ginseng Radix inhibits the DAG which is produced by the activation of mast cells with antigen-antibody reactions via both phosphatidylinositide-PLC and phosphatidylcholine-PLD systems, and then followed by the inhibition of histamine release. However, Re increases histamine release by stimulation of DAG production, which is mediated by phosphatidylcholine-PLD system rather than by phosphatidylinositide-PLC system, but inhibits the mass DAG production. Thus, it could be inferred that other mechanisms play a role in the increase of histamine release during mast cell activation.

• Korean Journal of Food Science and Technology
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• v.26 no.2
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• pp.178-183
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• 1994

The fresh perilla seed and tile one-year stored perilla seed were solvent extracted for their oil. On the other hand, the fresh seed and the stored seed were germinated in the dark at $25{\sim}28^{\circ}C\;for\;2{sim}3$ days and then solvent extracted. The above four kinds of perilla oil, that is, the oil from the nongerminated and fresh seed(NFO), the oil from the nongerminated and one-year stored seed (NSO), the oil from the germinated and fresh seed(GFO), and the oil from the germinated and one-year stored seed(GSO) were analyzed with regards to the chemical composition, and the effects of germination of the seed on the oxidative stability of perilla oil were studied. The iodine value and the saponification value were similar in all the perilla oils, but the acid value was increased by germination of the seed. The contents of free fatty acid and diacylglycerol were increased by germination of the seed, while the content of triacylglycerol was decreased. Of the polar lipid components, the content of phosphatidyl ethanolamine was greatly increased by germination of the seed. The contents of total tocopherol of perilla oil from the fresh seed and the one-year stored seed were 494 ppm and 439 ppm, respectively, and by germination of the seed increased to 560 ppm in GFO and 515 ppm in GSO, respectively. Especially a great change in the content of ${\gamma}-tocopherol$ was observed. The oxidative stability of perilla oil was increased by germination of the seed and the increase was distinct in the case of the one-year stored seed compared with that in the case of the fresh seed.

• Korean Journal of Food Science and Technology
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• v.26 no.5
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• pp.567-572
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• 1994

Diglyceride was prepared by reaction of hydrogenated beef tallow and glycerol (GL) in the presence of a Pseudomonas lipase. Both substrates were mixed at the ratio of GL/Triglyceride of 0.5 which is the stoichiometric molar ratio for the complete conversion of triglyceride (TG) to diglyceride (DG). DG can be obtained by solid phase-glycerolysis of hydrogenated beef tallow without use of organic solvents or emulsifiers by careful control of reaction temperature. Optimized reaction temperature condition was as follows: An initial incubation at$60^{\circ}C$ for 2h followed by the first temperature shift down to $55^{\circ}C$ for 4h, and then the second shift down to $50^{\circ}C$ for up to 3 days. There was a large decrease in the content of TG during the first $60^{\circ}C$ incubation for 2h. Even a prolonged incubation at $60^{\circ}C$ could not make a change of the composition of the reaction mixture at liquid state. By controlling the temperature lower than $60^{\circ}C$, reaction mixtures were solidified. The reaction temperature at $50^{\circ}C$ below the melting temperature of hydrogenaed beef tallow gave an 71% optimum yield of DG after 72h enzymatic glycerolysis and about 73% of total DG was 1,3-DG.

• Korean Journal of Food Science and Technology
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• v.36 no.5
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• pp.717-722
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• 2004

Response surface methodology was used to optimize production conditions of monoacylglycerol (MAG) and diacylglycerols (DAG) from corn oil by enzymic glycerolysis. Contents of $1,3-DAG\;(Y_1),\;1,2-DAG\;(Y_2),\;total\;DAG\;(Y_3),\;MAG\;(Y_4)$, and total $DAG+MAG\;(Y_5)$ were obtained. Conditions were optimized using central composite design with incubation temperature $(35-75^{\circ}C,\;X_1)$, incubation time (1-11 hr, $X_2$), and amount of hexane added (0-2 mL, $X_3$) as three variables. Content of 1,3-DAG was maximized by 20.43 area% at incubation temperature of $44.92^{\circ}C$, incubation time of 10.24 hr, and hexane content of 1.16 mL, whereas that of 1,2-DAG (26.78 area%) was maximized at $56.32^{\circ}C$, 6.95 hr, and 1.04 mL, respectively. Predicted maximum total DAG content was 45.09 area% at $53.82^{\circ}C$, 8.03 hr, and 1.08 mL, while production conditions of MAG (9.57 arae%) were $64.14^{\circ}C$, 7.00 hr, and 0.13 mL. At variables of $54.07^{\circ}C$, 7.98 hr, and 1.02 mL, maximum content of total DAG+MAG predicted by RSM was 53.54 area%.