• Nutritional Sciences
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• 제4권1호
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• pp.3-12
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• 2001

Long chain polyunsaturated fatty acids (LCPUFA) are important components of brain phospholipds and play important role (s) in brain function. In rats, the maximum brain growth occurs during the period of lactation even though it happens during the third trimester of gestation in human. Since milk contained docosahexaenoic acid (DHA) even through the maternal diet had no DHA and/or a very small amount of its precursor, $\alpha$-linolenic acid ($\alpha$-LnA), an emphasis was given to maternal adipose tissue as a reservoir of this fatty acid. We, therefore, investigated the mesenteric and subcutaneous adipose tissues for their fatty acid composition in dams reared with different fat diets. Diets containing various amounts of $\omega$6 and $\omega$3 fatty acids were given to adult female rats (200-250g) throughout the pregnancy and lactation periods. Diets were composed of 10% (wt/wt) corn oil (CO), soybean oil (SO), perilla seed oil (PO) containing about 60% $\alpha$-LnA, or fish oil (FO) rich in eicosapentaenoic acid (EPA) and DHA. The fatty acid ompositions of mesenteric and subcutaneous fat were measured and evaluated at Day-2 and Day-15 after parturition. In general, major characteristics of dietary fatty acid composition was reflected on the fatty acid composition of adipose tissues. Dietary fatty acid composition was reflected more on mesenteric fat as compared to subcutaneous fat. Mesenteric fat was found to contain less arachidonic acid (AA) and mesenteric fats of CO, SO and PO groups contained less DHA than did the subcutaneous fat. The P/M/S ratios of adipose tissues were similar between experimental groups while dietary P/M/S ratios differed significantly. It was noticeable that a small proportion of DHA was found in the adipose tissues of animals of CO, SO and PO groups (Day-2) and in SO and PO groups (Day-15), the groups which do not contain DHA in their diets. The percentage of DHA in mesenteric fat o CO, SO and PO groups decreased as lactation continues, while the proportion of DHA in FO group increased. Adipose tissues of FO group had higher DHA/EPA ratio as compared to the diet. Considering the fact that the body contains a large amount of adipose tissues, our present finding suggests that the adipose tissue can serve as a reservoir of DHA for pregnant and lactating rats.

• Journal of Nutrition and Health
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• 제26권6호
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• pp.661-671
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• 1993

The change in fatty acid composition in brain tissue of the second generation rats(Sprague-Dawley strain) was studied using four different fat diets(Corn oil=CO, Soybean oil=SO, Perilla oil=PO, Fish oil=FO, 10% by Wt). The experimental diets were started from pregnancy in four different groups, each consisting of 9 rats. The seound generation rats were fed the same diet as their mothers. Animals were anesthetized with ether at 0, 3, 9 & 16 weeks of age. Whole brains were dissected out, brain tissues were, then, homogenized and lipids were extracted from brain tissues. The fatty acid compositions were measured after methylation by gas-liquid chromatography at 0, 3, 9 and 16 weeks of age of offspring. The changes in the relative concentrations of polyunsaturated fatty acids(PUFA) or more specifically docosahexaenoic acid(22 : 6, $\omega$3, DHA), the major $\omega$3 fatty acid component in rat brain at different age were similar to changes in the amount of DNA in brain tissue showing the maximum value during the lactation. The changes in saturated fatty acid(SFA) content showed a contrasting patten to those of PUFA, while monounsaturated fatty acid(MUFA) increased steadily throughout the experimental period. At birth, the relative concentrations of $\omega$3 series fatty acids the relative concentrations of PUFA, MUFA and SFA converged to very similar values respectively regardless of the dietary fatty acid compositions. In brain tissue, it is of value to note that while changes in relative concentrations of linoleic acid (18 : 2, $\omega$6, LA) and arachidonic acid(20 : 4, $\omega$6, AA) showed a precursor-product-like relationship, $\alpha$-linolenic acid(18 : 3, $\omega$3, $\alpha$-LnA) and DHA showed a different pattern. Even when the $\omega$3 fatty acid content in very low in maternal diet(CO), the second generation rat brain tissues appeared to secure DHA content, suggesting an essential role of this fatty acid in the brain. The fact that a large amount of $\alpha$-LnA in the maternal diet did not have a significant effect on the second generation rat brain $\alpha$-LnA content, indicated that DHA seemed essential component for the brain development in our experimental condition. In all groups, the relative content of $\alpha$-LnA in the brain tissues remained relatively constant throughout the experimental period at the very low level. The study of the specific concentrations and essential role(s) of DHA in each parts of brain tissue is needed in more details.

• 대한한의학방제학회지
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• 제26권3호
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• pp.207-221
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• 2018

Objectives : Present study investigated hepatoprotective effect of Haegan-jeon extract (HE) and tried to elucidate molecular mechanism involved. According to molecular mechanism, present study optimized herbal composition of HE (op-HE) and compared in vitro and in vivo hepatoprotective effects of op-HE to HE. Methods : For in vitro experiments, HepG2 cells were exposed to arachidonic acid (AA, $10{\mu}M$) and iron ($5{\mu}M$) for inducing oxidative stress. Cell viability, GSH contents, $H_2O_2$ production, mitochondrial membrane potential, immunoblot and reporter gene assay were performed to investigate cytoprotective effects and responsible molecular mechanisms. For in vivo experiments, hepatoprotective effect of HE and op-HE were assessed on $CCl_4-induced$ liver injury mice model. Results : HE pretreatment prevented AA+iron-mediated hepatocytes apoptosis. In addition, AA+iron-induced mitochondrial dysfunction, $H_2O_2$ production, glutathione depletion were reduced by HE pretreatment. In addition, nuclear factor erythroid 2-related factor 2 (Nrf2) phosphorylation, antioxidant response element (ARE)-driven reporter gene activity, and antioxidant genes expression were increased by HE. Based on reporter gene and MTT assays, we found that op-HE consisting three medicinal herbs also significantly increased transactivation of Nrf2 and reduced the AA+iron-mediated cytotoxicity. Moreover, in $CCl_4-induced$ liver injury mice model, HE-op had an ability to ameliorate $CCl_4-mediated$ increases in serum alanine transferase and aspartate aminotransferase activity, hepatic degeneration, inflammatory cell infiltration, and collagen deposition. Hepatoprotective effects of op-HE were comparable to those of HE. Conclusions : Present study suggests that op-HE as well as HE exhibit hepatoprotective effect against oxidative stress-mediated liver injury via Nrf2 activation.

• The Korean Journal of Physiology and Pharmacology
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• 제5권4호
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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.

• 대한본초학회지
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• 제30권4호
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• pp.57-64
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• 2015

Objectives : Oxidative stress is one of the most causes of hepatocyte injury. Gleditsia spina, the thorns ofGleditsia sinensisLam., has been known for its anti-cancer and anti-inflammatory effects in Korean medicine. The present study investigated hepatoprotective effect of Gleditsia spina water extract (GSE) against oxidative stress induced by arachidonic acid (AA) + iron in HepG2 cells.Methods : To investigate cytoprotective effect of GSE, cells were pretreated with GSE and then subsequently exposed to 10 μM AA for 12 h, followed by 5 μM iron. Cell viability was monitored by MTT assay, and expression of apoptosis-related proteins was examined by immunoblot analysis. To identify responsible molecular mechanisms, reactive oxygen species (ROS) production, GSH contents, and mitochondrial membrane potential were measured. In addition, effect of GSE on nuclear factor erythroid 2-related factor 2 (Nrf2) activation was determined by immunoblot and antioxidant response element (ARE)-driven reporter gene assays.Results : GSE pretreatment prevented AA + iron-mediated cytotoxicity in concentration dependent manner. In addition, ROS production, glutathione depletion, and mitochondrial impairment by AA + iron were significantly inhibited by GSE. Furthermore, GSE promoted translocation of Nrf2 to nucleus, which acts as essential transcription factor for induction of antioxidant genes. Increased nuclear Nrf2 that caused by GSE treatment promoted transcriptional activity of ARE. Finally, GSE up-regulated sestrin-2 which was widely recognized as target gene of Nrf2.Conclusions : This study demonstrates that GSE protects hepatocytes from oxidative stress via activation of Nrf2 signaling pathway.

• 생명과학회지
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• 제16권6호
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• pp.942-948
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• 2006

본 연구에서는 DHA가 뇌에 미치는 영향을 알아보는 방법으로 시간과 동물의 수를 줄일 수 있는 장점을 지난 인공 사육 동물 시스템의 활용을 위한 전 단계 실험으로 엄마 쥐의 식이 섭취기간이 신생 쥐의 뇌 및 위 내용물의 지방산 조성에 미치는 영향에 대해 검토해 보았다. 출생 직후 신생 쥐들의 위 내용물의 지방산 조성의 경우, 임신동물을 사용한 군(Pregnancy group)의 경우, 임신전 단계 사육 방법으로 사육한 군(Pre-pregnancy group)에 비하여 20:2n-6 함량은 낮았고 20:3n-6, 20:4n-6 및 총 n-6 지방산의 경우는 높은 함량을 보였다. DHA 함량의 경우, 임신동물의 신생 쥐의 위 내용물에서 25% 높게 나타났다(P<0.05). 신생 쥐들이 생후 10일 경과되었을 때, 22:4n-6 및 22:6n-3의 경우 임신동물을 사용한 군에서 각각 8% 및 8%로 낮았으나 유의적 차이는 없었다. 출생 직후 뇌의 지방산 조성을 비교해 본 결과, 총 포화 지방산과 총 monounsaturated 지방산의 경우, 임신동물을 사용한 군(Pregnancy group)에서 그 함량이 유의적으로 높았고 20:5n-3의 함량은 26%로 감소하였다(P<0.05). 나머지 지방산 함량에는 두 간에 유의적 차이를 관찰 할 수가 없었다. 신생 쥐들이 생후 10일이 되었을 때, 총 포화지방산의 함량에는 차이가 없었으나 총 monounsaturated의 경우, 임신동물을 사용한 군에서 감소하고 20:4n-6와 22:6n-3의 함량은 유의적으로 높았으나 20:5n-3의 함량은 47%로 감소하였다(P<0.05). 이들 신생 쥐들이 수유기간이 끝나는 생후 21일이 되었을 때, 지방산 함량에 있어서 두 군간에 유의적 차이를 살펴 볼 수가 없었다. 이상의 결과에서 알 수 있듯이 임신 전 단계 군에 비하여 뇌 및 위 내용물의 DHA 및 기타 n-3 지방산의 함량이 높았으나 이유기의 뇌의 지방산 조성에는 변화가 없었다. 이러한 결과는 임신 동물을 사용한 군의 경우 임신 전 사료에 DHA가 함유된 사료를 섭취한 결과에 기인하는 것으로 여겨지며 이러한 특정 상황을 고려해 볼 때 향후 인공 사육 시스템을 이용할 경우 지방산의 조성의 차이는 미미한 것으로 사료되며 뇌의 DHA 결핍 동물 생성을 위한 실험계에 임신동물 활용이 가능하다고 사료되어진다.

• Asian-Australasian Journal of Animal Sciences
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• 제14권7호
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• pp.1024-1050
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• 2001

Avian spermatozoa are characterised by high concentrations of polyunsaturated fatty acids (PUFAs), in particular docosatetraenoic (DTA, 22:4n-6) and arachidonic (AA, 20:4n-6) acids. As a result they are vulnerable to lipid peroxidation, which is considered to be an important factor of male infertility. Antioxidant systems are expressed in spermatozoa and seminal plasma and build three major levels of antioxidant defense. The first level is based on the activity of superoxide dismutase (SOD) which is, in conjunction with glutathione peroxidase (GSH-Px), catalase and metal-binding proteins, responsible for prevention of free radical formation. The second level of defence is responsible for prevention and restriction of chain reaction propagation and includes chain-breaking antioxidants such as vitamin E, ascorbic acid, glutathione and some others. The third level of antioxidant defence deals with damaged molecules, repairing or removing them from the cell and includes specific enzymes such as lipases, proteases, DNA repair enzymes etc. In the review, profiles of PUFAs and the two first lines of antioxidant defence in avian spermatozoa are characterised. Dietary manipulation of the breeder's diet (PUFA, vitamin E and selenium) as an effective means of modulating fatty acid composition and antioxidant system is also considered. Antioxidant properties of seminal plasma and efficiencies of inclusion of antioxidants into semen diluents are also characterised.

• Archives of Pharmacal Research
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• 제25권5호
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• pp.675-680
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• 2002

In order to investigate the underlying mechanism of HCI in oesophagitis, the inflammatory response to HCI was observed in RBL-2H3 mast cells. Rat basophilic leukemia (RBL-2H3) cells were used to measure histamine release, arachidonic acid (AA) release, reactive oxygen species (ROS) and peroxynitrite generation induced by HCI. Exogenous HCl increased the level of histamine release and ROS generation in a dose dependent manner, whereas it decreased the spontaneous release of [$^3$H] M and the spontaneous production of peroxynitrite. Mepacrine (10 $\mu$M), oleyloxyethyl phosphorylcholine (10 $\mu$M) and bromoenol lactone (10 $\mu$M) did not affect both the level of histamine release and ROS generation induced by HCI. U73122 (1 $\mu$M), a specific phospholipase C (PLC) inhibitor did not have any influence on level of histamine release and ROS generation. Propranolol (200 $\mu$M), a phospholipase D (PLD) inhibitor, and neomycin (1 mM), a nonspecific PLC and PLD inhibitor, significantly inhibited both histamine release and ROS generation. Diphenyleneiodonium (10 $\mu$M), a NADPH oxidase inhibitor, and tiron (5 mM), an intracellular ROS scavenger significantly inhibited the HCI-induced histamine release and ROS generation. These findings suggest that the inflammatory responses to HCI is related to histamine release and ROS generation, and that the ROS generation by HCI may be involved in histamine release via the PLD pathway in RBL-2H3 cells.

• 생명과학회지
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• 제17권4호
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• pp.546-551
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• 2007

본 연구에서는 n-3계 지방산이 풍부하게 함유된 고등어를 기존의 열풍건조기의 단점을 보완하고 높은 온도에서 손실되는 영양소를 막기 위해 저온진공건조기를 도입하여 건조시킨 후, 건조고등어의 섭취에 의한 간 및 신경조직에서의 지방산 조성 변화를 알아보고자 하였다. 간 지방산 조성의 경우, 고등어군의 총 n-6 지방산 함량은 두 군에서 유의적 차이는 없었으나 총 n-3 지방산의 함량은 고등어군에서 대조군보다 증가하였다. N-3 계열 지방산 중에서 특히 DHA의 함량은 대조군에 비해 229% 증가하였다(p<0.05). 대뇌 피질의 지방산 조성의 경우, 고등어군의 총 monounsaturated 지방산은 대조군에 비해 유의적으로 증가하였으나, 총 n-6 지방산의 함량은 감소하였다(p<0.05). 반면 총 n-3 지방산은 대조군에 비해 12%로 증가하였고 그 중에서 22:5n-3와 DHA의 함량은 고등어군에서 대조군보다 증가하였다(p<0.05). 소뇌의 지방산 조성의 경우, 총 지방산 함량과 총 n-6 지방산의 함량이 고등어군에서 대조군보다 감소하였고, 총 n-3 지방산의 함량은 두 군 간에 유의적 차이는 없었으나 n-3계 지방산중 22:5n-3의 함량은 200%로 증가하였다(p<0.05). 망막의 지방산 조성의 경우, 고등어군의 총 n-6 지방산의 함량은 대조군보다 감소하였으나, 총 n-3 지방산의 함량은 유의적 차이가 나타나지 않았다. 반면, n-3계 지방산 중에서 20:5n-3과 22:5n-3이 각각 200%, 67%로 증가하였다(p<0.05). 이상의 결과로부터 n-3 지방산을 많이 함유한 건조 고등어 식이는 간 및 신경조직의 총 n-3 지방산의 함량을 증가시켰고, 특히 간, 대뇌 피질에서의 DHA의 함량을 증가시켰다. 이상의 결과를 토대로 건조 고등어 섭취는 뇌의 DHA 함량을 증가시켰으므로 향후 공간 기억력 테스트를 행하여 고등어 섭취에 의한 뇌 기능 개선 효과와 뇌의 DHA 함량 증가와의 상호 연관성에 대하여 검토할 예정이다.

• 동의생리병리학회지
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• 제19권5호
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• pp.1391-1398
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• 2005

The anti-thrombic properties of the oriental herbal medicine Daejowhan(DJW, 大造丸) which consists of 11 kinds of herbs (indicated as ratio) of Rehmanniae Radix 24%, Hominis Placenta 5%, Testudinis Carapax 9%, Eucommiae Cortex 9%, Asparagi Radix 9%, Phellodendri Cortex 9%, Achyranthis Radix 7%, Liriopis Tuber 7%, Angelicae Sinensis Radix 7%, Ginseng Radix 5% and Schizandrae Fructus 3% were investigated. The water extracts from DJW inhibited Platelet-activating factor(PAF) induced platelet aggregation. DJW was extracted with methanol and further fractionated by ethylacetate. A 70% methanol extract showed a strong inhibition against PAF-induced aggregation in vitro and in vivo assays. The ethylacetate soluble fraction was shown to have inhibitory effect on PAF-induced platelet aggregation in vitro assay. The ethylacetate soluble fraction specially protected against the lethality of PAF, while verapamil did not afford any protection. These results indicate that the water extracts and alcoholic-fractions inhibit the action of PAF in vivo by an antagonistic effect on PAF, so that it may be useful in treating disorders caused by PAF, such as acute allergy, inflammation, asthma, gastrointestinal ulceration, toxic shock and so forth. DJW was investigated regarding its assumed anti-thrombic action on human platelets which was deduced from its ability to suppress Arachidonic acid(AA)-induced aggregation, exocytosis of ATP, and inhibition of Cyclooxygenase(COX) and Thromboxane synthase(TXS) activity. The latter two effects were estimated from the generation of Prostaglandin $E_2(PGE_2)$ and Thromboxane $A_2(TXA_2)$ respectively. Exogenously applied AA ($100{\mu}mol/{\ell}$) provoked a $89\%$ aggregation of platelets, the release of 14 pmol ATP, and the formation of either 225 pg $TXA_2$ or 45 pg $PGE_2$, each parameter being related to 106 platelets. An application of DJW 5 min before AA dose-dependently diminished aggregation, ATP-release and the synthesis of $TXA_2$ and $PGE_2$ with $IC_{50}$ values of 74, 108, 65, $72{\mu}g/m{\ell}$, respectively. The similarity of the $IC_{50}$ values suggest an inhibition of COX by DJW as primary target, thus suppressing the generation of $TXA_2$ which induces aggregation of platelets and exocytosis of ATP by its binding on $TXA_2$-receptors.