• Title/Summary/Keyword: cPLA2

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Hydrolysis of Phosphatidyicholine to Initiate HeLa Cell Adhesion to a Gelatin Substratum (Phosphatidylcholine의 분해에 의한 Hela 세포와 Gelatin 기질과의 상호작용의 유도)

  • ;;;;;Bruce S. Jacobson
    • The Korean Journal of Zoology
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    • v.38 no.4
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    • pp.457-464
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    • 1995
  • Hela cells, a transformed human epithelial cell line, attach to various substrata but subsequent spreading is specific to collagen or gelatin. The spreading is initiated by the activation of phospholipase $A_2$ (PLA$_2$) which produces arachidonic acid (AA) as a consequence of cell surface collagen receptor clustering. This study examines the mechanism of PLA$_2$activation and which phospholipids are hydrolyzed by PIA$_2$ to release AA in response to Hela cell adhesion to a gelatin substratum. The levels of phosphatidyicholine decreases, among various phospholipids, during attachment and spreading of Hela cells. Lysophosphatidyicholine Is the only lysophospholipids formed during ileLa cell adhesion indicating that clustered collagen receptors activate PLA$_2$to hydrolyze posphatidylcholine to AA and lysophosphatidylcholine. Among various molecular entitles which are known to regulate PLA$_2$ activation, we have previously shown that PLA2 activation is not mediated by either changes in $Ca_2$+ levels, alkalinization of cytoplasmic p11, or activation of protein hinase C. It is also likely that PIA2 activation is not mediated by either pertussis or cholera toxinsensitive G proteins as those toxins do not affect both AA release and cell spreading.

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Involvement of Cytosolic Phospholipase $A_2$ in Nerve Growth Factor-Mediated Neurite Outgrowth of PC12 Cells

  • Choi, Soon-Wook;Yu, Eun-Ah;Lee, Young-Seek;Yoo, Young-Sook
    • BMB Reports
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    • v.33 no.6
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    • pp.525-530
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    • 2000
  • The nerve growth factor (NGF) induces neuronal differentiation and neurite outgrowth of PC12 cells, whereas epidermal growth factors (EGF) stimulate growth and proliferation of the cells. In spite of this difference, NGF-or EGF-treated PC12 cells share various properties in cellular-signaling pathways. These include the activation of the phosphoinositide (PI)-3 kinase, 70 kDa S6 kinase, and in the mitogen-activated protein (MAP) kinase pathway, following the binding of these growth factors to intrinsic receptor tyrosine kinases (RTKs). Therefore, many studies have been attempted to access the critical signaling events in determining the differentiation and proliferation of PC12 cells. In this study, we investigated the cytosolic phospholipase $A_2$ ($cPLA_2$) in neurite behavior in order to identify the differences of signaling pathways between the NGF-induced differentiation and the EGF-induced proliferation of PC12 cells. We have showed here that the $cPLA_2$ was translocated from cytosol to membrane only in NGF-treated cells. We also demonstrated that this translocation is associated with NGF-induced activation of phospholipase $C-{\gamma}(PLC-{\gamma})$, which elevates intracellular $Ca^{2+}$ concentration. These results reveal that the translocation of $cPLA_2$ may be a requisite event in the neuronal differentiation of PC12 cells. Various phospholipase inhibitors were used to confirm the importance of these enzymes in the differentiation of PC12 cells. Neomycin B, a PLC inhibitor, dramatically inhibited the neurite outgrowth, and two distinct $PLA_2$ inhibitors, 4-bromophenacyl bromide (BPB) and arachidonyltrifluoro-methyl ketone ($AACOCF_3$) also suppressed the neurite outgrowth of the cells, as well Taken together, these data indicated that $cPLA_2$ is involved in NGF-induced neuronal differentiation and neurite outgrowth of PC12 cells.

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Effect of Cellular Phospholipase A2 Inhibition on Enhancement of Bt Insecticidal Activity (세포성 인지질분해효소 활성 억제에 따른 비티 살충력 증가 효과)

  • Eom, Seonghyeon;Park, Jiyeong;Kim, Kunwoo;Kim, Yonggyun
    • Korean journal of applied entomology
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    • v.53 no.3
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    • pp.271-280
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    • 2014
  • Some bacterial metabolites of Xenorhabdus nematophila (Xn) inhibit phospholipase $A_2$ ($PLA_2$) activity to shutdown eicosanoid biosynthesis in target insects. However, little has been known about the target insect $PLA_2$ of these bacterial metabolites. Eight bacterial metabolites identified in Xn culture broth exhibited significant insecticidal activities against larvae of both lepidopteran species of Plutella xylostella and Spodoptera exigua. Moreover, these bacterial metabolites significantly enhanced insecticidal activities of Bacillus thuringiensis (Bt). To determine target $PLA_2$, we cloned and over-expressed cellular $PLA_2$ ($SecPLA_2$) of S. exigua. Purified $SecPLA_2$ catalyzed phospholipids derived from the fat body and released several polyunsaturated fatty acids. Most Xn metabolites significantly inhibited $SecPLA_2$ activity, but were different in their inhibitory activities. There was a positive correlation between the inhibition of $SecPLA_2$ and the enhancement of Bt insecticidal activity. These results indicate that $SecPLA_2$ is a molecular target inhibited by Xn metabolite.

Effect of In Vitro Degradation on the Weight Loss and Tensile Strength of PLA/PEG Melt Blend Fiber (In Vitro 분해가 PLA/PEG 용융블렌드 섬유의 무게감량률 및 인장강도에 미치는 영향)

  • Yoon, Cheol-Soo;Ji, Dong-Sun
    • Polymer(Korea)
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    • v.33 no.6
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    • pp.581-587
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    • 2009
  • PLA/PEG blend fibers composed of poly (lactic acid) (PLA) and poly (ethylene glycol) (PEG) were prepared via melt blending and spinning for bioabsorbable filament sutures. The blend fibers hydrolyzed with the immersion in a phosphate buffer solution at pH 7.4 and $37\;^{\circ}C$ for 1~8 weeks. The effects of blending time, blend composition, and hydrolysis time on the weight loss and tensile strength of the hydrolyzed blend fibers were investigated. After hydrolysis, the weight loss of the blend fibers increased with increasing PEG content, blending time, and hydrolysis time. The tensile strength and tensile modulus of the blend fibers decreased with increasing PEG content, blending time, and hydrolysis time. Therefore, it can be concluded that the weight loss of the PLA/PEG blend fibers was less than 0.9% even at hydrolysis time of 2 weeks and their strength retentions were over 90%.

Toughening of PLA stereocomplex by Impact modifiers (충격보강제에 의한 PLA stereocomplex의 강인화 연구)

  • Nam, Byeong-Uk;Lee, Bum-Suk
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.13 no.2
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    • pp.919-925
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    • 2012
  • We tried to blend PLLA and PDLA at overall compositions to form PLA stereocomplexes (SC). The presence of the SC crystalline phase in the PLLA matrix was verified by differential scanning calorimetry (DSC). As a result, a various PDLA composition of the PLA SC blends can influence PLA SC formation. And the largest amount of PLA SC crystallites was formed when PLLA/PDLA ratio is 50/50. In addition, we have tried to do PLA SC toughening with two impact modifiers in 92/8, 85/15 ratio of PLLA/PDLA to enhance the mechanical properties such as impact strength. Thermal and mechanical properties of PLA SC were investigated by DSC, HDT, Izod impact tester and UTM. PLA SC formation decreased when 10-20 wt% of Strong120 (impact modifier) was added. On the other hand, there is no effect on PLA SC formation when 10-20% of Elvaloy (impact modifier) was added. HDT values dramatically increased over $100^{\circ}C$ with the addition of PDLA. However, HDT decreased as Strong120 and Elvaloy content increased. Finally, we could find well balanced composition of toughened PLA SC with 10wt% of impact modifier in flexural modulus and impact strength.

Characteristics of Cytosolic Calcium-Independent Phospholipase $A_2$ Isolated from Rat Liver

  • Won, Jong-Hak;Na, Doe-Sun;Rhee, Hae-Jin;Park, Young-Min
    • BMB Reports
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    • v.32 no.2
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    • pp.154-160
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    • 1999
  • A calcium-independent phospholipase $A_2$ ($iPLA_2$) was identified from the cytosolic fraction of rat liver cells. On gel filtration chromatography, the $iPLA_2$ activity was eluted as broad peaks of 150 to 500 kDa. The enzyme was maximally active at pH 7.5, retained 75% of its original activity after heating at $50^{\circ}C$ for 5 h, and was inhibited by $Ca^{2+}$, $Mg^{2+}$, and $Zn^{2+}$ ions, but was not affected by $Na^+$ and $K^+$ ions. The enzymatic activity was increased up to 150% by 1 to 4 mM DTT and was inhibited up to 25% by 0.1 to 1 mM PMSF. The $iPLA_2$ activity had preference for the head group of phospholipids, where phosphatidylethanolamine was preferred to phosphatidylcholine. The results suggest that the $iPLA_2$ may be a novel enzyme distinct from the previously reported $iPLA_2s$.

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Role of Group II Phospholipase $A_2$ in the Pulmonary Oxidative Stress of the Acute Lung Injury Induced by Gut Ischemia-Reperfusion (장의 허혈-재관류로 유도된 급성 폐손상에서 산화성 스트레스에 관여하는 group II phospholipase $A_2$의 역할)

  • Jheon, Sang-Hoon;Kim, Keun;Lee, Sang-Cheol;Kim, Seong-Eun;Lee, Young-Man;Lee, Jong-Tae
    • Journal of Chest Surgery
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    • v.35 no.7
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    • pp.501-510
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    • 2002
  • Background: The various pathogeneses of acute respiratory distress syndrome have been suggested but not established yet. In the present study, the role of group II phospholipase $A_2$($PLA_2$) in the pathogenesis of gut ischemia-reperfusion(I/R) induced acute lung injury (ALI), especially in the pulmonary oxidative stress with infiltration of neutrophils was investigated. Material and Method: To induce ALI, reperfusion of mesentery was done for 120 min after clamping of superior mesenteric artery for 60 min in Sprague-Dawley rats that weighed about 300g. To exmaine the role of group II $PLA_2$ in ALI, especially endothelial injury associated with the action of neutrophils, lung myeloperoxidase activity, lung leak index, bronchoalveolar lavage fluid protein were measured, and pulmonary $PLA_2$ activity changes in gut I/R were also measured. The role of group II $PLA_2$in the neutrophilic generation of free radicals was assessed by inhibiting group II $PLA_2$ with rutin, manoalide and scalaradial. Furthermore, to verify the oxidative stress in the lung, histologic and free radical detecting cytochemical electron microscopy were done. Result: After reperfusion, ALI was developed with accumulation of neutrophils in the lung, which was confirmed by the increase of myeloperoxidase activity, lung leak index and bronchoalveolar lavage protein (p<0.001). The pulmonary and intestinal group II $PLA_2$ activities significantly increased after gut I/R which were reversed by rutin(p<0.001). In vitro, cytochrome-c reduction assay denoted the inhibitory effects of rutin, scalaradial and manoalide on the production of free radicals from isolated human neutrophils. Histologically, neutrophilic accumulation and pericapillary edema in the lung after gut I/R was detected by light microscopy which was suppressed by rutin. In $CeCl_3$ cytochemical electron microscopy, the increased production of hydrogen peroxide in the lung after gut I/R was confirmed and also the production of hydrogen peroxide was decreased by rutin. Conclusion: On the basis of these experimental results, the inhibition of group II $PLA_2$ seemed to mitigate gut I/R-induced ALI by suppressing the production of free radicals from the infiltrated neutrophils. Collectively, group II $PLA_2$ seems to play a crucial role in gut I/R-induced ALI by neutrophilic oxidative stress.

Two Phenolic Compounds Isolated from Umbilicaria esculenta as Phospholipase $A_2$ Inhibitors (석이로부터 분리한 페놀성 화합물의 phospholipase $A_2$ 저해활성)

  • Kim, Jin-Woo;Song, Kyung-Sik;Yoo, Ick-Dong;Chang, Hyeun-Wook;Yu, Seung-Hun;Bae, Kang-Gyu;Min, Tae-Jin
    • The Korean Journal of Mycology
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    • v.24 no.3 s.78
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    • pp.237-242
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    • 1996
  • Two depsides, medicinal herb products isolated from the methanol extract of Umbilicaria esculenta, inhibited human synovial fluid Phospholipase $A_2\;(PLA_2)$ ($IC_{50}$ of 0.22 and 0.26 mM, respectively). In the course of screening for antiinflammatory compounds from natural products, we successfully isolated two depsides $PLA_2$ inhibitory compounds, Orcinol and methyl orsellinic acid. The compounds were identified as orcinol and methyl orsellinic acid on the basis of various NMR studies including $^1H,\;^{13}C$ and DEPT experiments.

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Arachidonic Acid Activates $K^+$-$Cl^-$-cotransport in HepG2 Human Hepatoblastoma Cells

  • Lee, Yong-Soo
    • The Korean Journal of Physiology and Pharmacology
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    • v.13 no.5
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    • pp.401-408
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    • 2009
  • $K^+$-$Cl^-$-cotransport (KCC) has been reported to have various cellular functions, including proliferation and apoptosis of human cancer cells. However, the signal transduction pathways that control the activity of KCC are currently not well understood. In this study we investigated the possible role of phospholipase $A_2$ ($PLA_2$)-arachidonic acid (AA) signal in the regulatory mechanism of KCC activity. Exogenous application of AA significantly induced $K^+$ efflux in a dose-dependent manner, which was completely blocked by R-(+)-[2-n-butyl-6,7 -dichloro-2-cyclopentyl-2,3-dihydro-1-oxo-1Hinden-5-yl]oxy]acetic acid (DIOA), a specific KCC inhibitor. N-Ethylmaleimide (NEM), a KCC activatorinduced $K^+$ efflux was significantly suppressed by bromoenol lactone (BEL), an inhibitor of the calciumindependent $PLA_2$ ($iPLA_2$), whereas it was not significantly altered by arachidonyl trifluoromethylketone ($AACOCF_3$) and p-bromophenacyl bromide (BPB), inhibitors of the calcium-dependent cytosolic $PLA_2$ ($cPLA_2$) and the secretory $PLA_2$ ($sPLA_2$), respectively. NEM increased AA liberation in a doseand time-dependent manner, which was markedly prevented only by BEL. In addition, the NEM-induced ROS generation was significantly reduced by DPI and BEL, whereas $AACOCF_3$ and BPB did not have an influence. The NEM-induced KCC activation and ROS production was not significantly affected by treatment with indomethacin (Indo) and nordihydroguaiaretic acid (NDGA), selective inhibitors of cyclooxygenase (COX) and lipoxygenase (LOX), respectively. Treatment with 5,8,11,14-eicosatetraynoic acid (ETYA), a non-metabolizable analogue of AA, markedly produced ROS and activated the KCC. Collectively, these results suggest that $iPLA_2$-AA signal may be essentially involved in the mechanism of ROS-mediated KCC activation in HepG2 cells.

Effects of In Vitro Degradation on the Weight Loss and Tensile Properties of PLA/LPCL/HPCL Blend Fibers

  • Yoon Cheol Soo;Ji Dong Sun
    • Fibers and Polymers
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    • v.6 no.1
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    • pp.13-18
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    • 2005
  • PLA/LPCL/HPCL blend fibers composed of poly (lactic acid) (PLA), low molecular weight poly ($\varepsilon$-caprolactone) (LPCL), and high molecular weight poly ($\varepsilon$-caprolactone) (HPCL) were prepared by melt blending and spinning for bioab­sorbable filament sutures. The effects of blending time and blend composition on the X-ray diffraction patterns and tensile properties of PLA/LPCL/HPCL blend fibers were characterized by WAXD and UTM. In addition, the effect of in vitro degra­dation on the weight loss and tensile properties of the blend fibers hydrolyzed during immersion in a phosphate buffer solu­tion at pH 7.4 and 37$^{\circ}C$ for 1-8 weeks was investigated. The peak intensities of PLA/LPCL/HPCL blend fibers in X-ray diffraction patterns decreased with an increase of blending time and LPCL contents in the blend fibers. The weight loss of PLA/LPCL/HPCL blend fibers increased with an increase of blending time, LPCL contents, and hydrolysis time while the tensile strength and modulus of the blend fibers decreased. The tensile strength and modulus of the blend fibers were also found to be increased with an increase of HPCL contents in the blend fibers. The optimum conditions to prepare PLA/LPCL/HPCL blend fibers for bioabsorbable sutures are LPCL contents of $5 wt\%, HPCL contents of $35 wt\%, and blending time of 30 min. The strength retention of the PLA/LPCL/HPCL blend fiber prepared under optimum conditions was about $93.5\% even at hydrolysis time of 2 weeks.