인간 추간판의 섬유륜부 및 수핵부 세포 배양에서 세포의 형질에 따른 phosphollpase A2 의 생성

김동준,왕진만 대한척추외과학회 2000 대한척추외과학회지 Vol.7 No.1

연구계획 : 추간판 세포의 배양에서 세포의 형질에 따른 phospholipas A2의 생성 정도의 차이를 규명한다. 연구목적 : 통증의 발생 기전에서 중요한 역할을 하는 phospholipase A2의 생성과 연관하여, 주로 관계하는 세포 형질과 생성을 촉진하는 조건에 대해 알아보고자 한다. 대상 및 방법 : 변성되지 않은 인체의 추간판에서 외연의 섬유륜부와 중심의 수핵부에서 각각 조직을 채취하여 세포를 배양한다. 배양 7일째에 배양된 세포들을 조직별로 3군씩 나누고 젖산을 0mM(대조군), 1mM, 5mM을 첨가하여 2주간 더 배양한 다음 phospholipase A2의 생성 정도를 Northerm blotting으로 측정한다. 결과 : 수핵 조직에서 채취된 세포들에서는 적은 양의 phospholipase A2가 생성되었다. 반면에 섬유륜 조직의 세포들은 왕성한 phospholipase A2의 생성을 보였다. 첨가된 젖산의 양에 따른 phospholipase A2의 생성 정도는 뚜렷한 차이가 없었다. 결론 : 이런 결과로 볼 때 추간판을 구성하는 조직들 중에서 외연의 섬유륜부 세포들이 phospholipase A2의 생성에 주로 관계하며, 추간판성 통증의 발생에 중요한 역할을 하는 것으로 생각된다. Study Design : Evaluation of phospholipase A2 production according to cell type of human intrevertebral disc. Summary of Literature Review : It was reported that the phospholipase A2 acitvity in human lumbar desc herniation was more active than that in other tissues. Objectives : The purpose of this study was to evaluate the diferences between the cells of anulus fibrosus and nucleus pulposus when lactate was added to the culture medium. Materials and Methods : Cells from the anulus fibrosus and nucleus pulposus of a human intervertebral disc were prepared enzymaitically. After the monolayer was set up, the cells were didvided to three groups and lactate doses of a OmM, 2mM or 5mM were added respectively. At two week after lactate addition the production of phospholipase A2 was measured by Northerm blotting. Results : Cells of nucleus pulposus produced a small amount of phospholipase A2. Those of anulus fibrosus showed a high activity of phospholipase A2 production. The concentration of lactate did not influenced on the production of phospholipase A2. Conclusion : The anulus fibrosus hsa an improtant role in the production of phospholipase A2 and is thought to be related with generation of discogenic pain.

Modulation of Uterine Phospholipase $A_2$ Activity by Estradiol During the Delayed Implantation Process in Rats

윤미정,김창미,최임순,유경자,Yoon, Mi-Chung,Kim, Chang-Mee,Choe, Rim-Soon,Ryu, Kyung-Za The Korean Society of Pharmacology 1991 대한약리학잡지 Vol.27 No.2

본 연구에서는 흰쥐의 수정란 착상시기에 estradiol이 prostaglandins(PGs) 합성의 전구체인 arachidonic acid를 생성하는데 관여하는 phospholipase $A_2(PLA_2)$의 활성도를 조절하므로써 PGs의 합성을 촉진하는가를 조사하여 다음과 같은 결과를 얻었다. 자궁의 $PLA_2$ 활성도는 수정란이 착상하는 시기인 임신 제 5일에 증가되었으며, 비착상부위에서보다는 착상부위에서 더 높은 것으로 나타났다. Delayed implantation model에서, $PLA_2$ 활성도는 estradiol을 투여한 지 11시간후에 증가되었으며, dbcAMP를 투여한지 8시간후에 증가되었다. 또한 estradiol을 투여하기 2시간전에 phosphodiesterase inhibitor인 theophylline을 투여하면 estradiol만 투여한것에 비하여 $PLA_2$ 활성도가 증가되었다. Estradiol 또는 dbcAMP와 함께 indomethacin을 투여하면 자궁의 PGs합성은 억제되었으나 $PLA_2$ 활성도에는 영향을 주지않았다. 이상의 결과로 보아 흰쥐의 착상시기에 estradiol은 cAMP를 매개로하여 자궁의 $PLA_2$ 활성도를 촉진하므로써 PGs의 합성을 증가시키는 것으로 생각된다. The present study was performed to determine whether estradiol, via cAMP mediation, induces prostaglandin synthesis by modulating phospholipase $A_2$ activity which hydrolyzes phospholipids into arachidonic acids, a precursor for prostaglandin synthesis, during the implantation process in rats. Uterine phospholipase $A_2$ activity was elevated on day 5 of pregnancy when implantation normally occurs in rats. Moreover, phospholipase $A_2$ activity was higher in the implant sites than in the non-implant sites of uterus on day 6. In delayed implantation model, phospholipase $A_2$ activity was increased at 12 hrs after estradiol administration and at 8 hrs after dbcAMP administration. In addition, higher activity of phospholipase $A_2$ was induced by the treatment of estradiol plus theophylline, compared with estradiol-only treated group. The simultaneous treatment of indomethacin with estradiol or dbcAMP did not alter phospholipase $A_2$ activity compared with estradiol or dbcAMP-only treated group although significant suppression was observed in uterine PGE and $PGF_{2{\alpha}}$ concentrations. These results suggest that estradiol or cAMP stimulates uterine phospholipase $A_2$ activity, thereby increasing prostaglandin synthesis during the implantation process in rats.

흰쥐의 착상기간중 Estradiol이 자궁의 Phospholipase A₂ 활성도에 미치는 영향

윤미정(Mi-chung Yoon),김창미(Chang-mee Kim),최임순(Rim-Soon Choe),유경자(Kyung-za Ryu) 대한약리학회 1991 대한약리학잡지 Vol.27 No.2

본 연구에서는 흰쥐의 수정란 착상시기에 estradiol이 prostaglandins(PGs) 합성의 전구체인 arachidonic acid를 생성하는데 관여하는 phospholipase A₂(PLA₂)의 활성도를 조절하므로써 PGs의 합성을 촉진하는가를 조사하여 다음과 같은 결과를 얻었다. 자궁의 PLA₂ 활성도는 수정란이 착상하는 시기인 임신 제 5일에 증가되었으며, 비착상부위에서보다는 착상부위에서 더 높은 것으로 나타났다. Delayed implantation model에서, PLA₂ 활성도는 estradiol을 투여한 지 11시간후에 증가되었으며, dbcAMP를 투여한지 8시간후에 증가되었다. 또한 estradiol을 투여하기 2시간전에 phosphodiesterase inhibitor인 theophylline을 투여하면 estradiol만 투여한것에 비하여 PLA₂ 활성도가 증가되었다. Estradiol 또는 dbcAMP와 함께 indomethacin을 투여하면 자궁의 PGs합성은 억제되었으나 PLA₂ 활성도에는 영향을 주지않았다. 이상의 결과로 보아 흰쥐의 착상시기에 estradiol은 cAMP를 매개로하여 자궁의 PLA₂ 활성도를 촉진하므로써 PGs의 합성을 증가시키는 것으로 생각된다. The present study was performed to determine whether estradiol, via cAMP mediation, induces prostaglandin synthesis by modulating phospholipase A₂ activity which hydrolyzes phospholipids into arachidonic acids, a precursor for prostaglandin synthesis, during the implantation process in rats. Uterine phospholipase A₂ activity was elevated on day 5 of pregnancy when implantation normally occurs in rats. Moreover, phospholipase A₂ activity was higher in the implant sites than in the non-implant sites of uterus on day 6. In delayed implantation model, phospholipase A₂ activity was increased at 12 hrs after estradiol administration and at 8 hrs after dbcAMP administration. In addition, higher activity of phospholipase A₂ was induced by the treatment of estradiol plus theophylline, compared with estradiol-only treated group. The simultaneous treatment of indomethacin with estradiol or dbcAMP did not alter phospholipase A₂ activity compared with estradiol or dbcAMP-only treated group although significant suppression was observed in uterine PGE and PGE_2α concentrations. These results suggest that estradiol or cAMP stimulates uterine phospholipase A₂ activity, thereby increasing prostaglandin synthesis during the implantation process in rats.

사람 양수중 다종의 세포외성 포스포리파제 A2의 부분정제 및 특성

전용주(Yong Ju Jeon),백석환(Suk Hwan Baek),이지혜(Jee Hae Lee),문태철(Tae Chul Moon),민병우(Beong Woo Min),장현욱(Hyeun Wook Chang) 대한약학회 1997 약학회지 Vol.41 No.2

Multiple forms of extracellular phospholipase A2 have been detected in human amniotic fluid (HAF). When HAF was subjected to heparin-Sepharose column chromatography, phospholipase A2 activity was detected in both heparin-non binding and binding fraction. The activity of heparin-non binding fraction was further purified by sequential uses of column chromatographies on butyl-Toy-opearl 650M and DEAE-Sephacel. DEAE-Sephacel fraction contained three different phospholipase A2 activities (Peak I, II, III). The molecular weight of DEAE-Sephacel fraction phospholipase A2 determined by SDS-PAGE were about 52KDa (Peak I). Peak II, III required micromolar Ca2+ ion for its maximum activity, but Peak I enzyme showed calcium independent phospholipase A2 activity and showed broad range of pH (6.0~10.0) optimum. All these enzymes were not recognized by a monoclonal antibody raised against phospholipase A2 from human synovial fluid. These results suggest that HAF might contain multiple forms of extracellular phospholipase A2, which may neither belong to the 14KDa group II phospholipase A2 family nor cytosolic phospholipase A2.

약침용(藥鍼用) 봉독성분(蜂毒成分) 중(中) Apamin, Melittin의 항암작용(抗癌作用)

권도희,이재동,최도영,Kwon, Do-Hee,Lee, Jae-dong,Choi, Do-Yong 대한침구의학회 2001 대한침구의학회지 Vol.18 No.1

Objectives : To characterize the antitumorigenic potential of three representative bee venom components, Melittin, Apamin, and Phospholipase A2, their effects on cell proliferation and apotosis of the human melanoma cell line SK-MEL-2 were analyzed using molecular biological approaches. Methodes & Results : To determine the doses of the drugs that do not induce cytotoxic damage to this cell line, cell viability was examined by MTT assay. While SK-MEL-2 cells treated with 0.5 - 2.0㎍/㎖ of each drug showed no recognizable cytotoxic effect, marked reductions of cell viability were detected at concentrations over 5.0㎍/㎖. [3H]thymidine incorporation assay for cell proliferation demonstrated that DNA replication of SK-MEL-2 cells is inhibited by Apamin and Phospholipase A2 in a dose-dependent manner. Consistent with this result, the cells were accumulated at the G1 phase of the cell cycle after treatment with Apamin and Phospholipase A2, whereas no detectable change in cell proliferation was identified by Melittin treatment. In addition, tryphan blue exclusion and flow cytometric analyses showed that all of these drugs can trigger apoptotic cell death of SK-MEL-2, suggesting that Melittin, Apamin, and Phospholipase A2 have antitumorigenic potential through the suppression of cell growth and/or induction of apoptosis. Qantitative RT-PCR analysis revealed that Apamin and Phospholipase A2 inhibit expression of growth-promoting genes such as c-Jun, c-Fos, and Cyciin D1. Furthermore, Phospholipase A2 induced tumor suppressors p53 and p21/Wafl. In addition, all three drugs were found to activate expression of a representative apoptosis-inducing gene Bax while expression of apoptosis-suppressing Bcl-2 and Bcl-XL genes was not changed. Taken together, this study strongly suggests that Metittin, Apamin, and Phosphalipase A2 may have antitumorigenic activities, which are associated with its growth-inhibiting and/or apoptosis-inducing potentials.

A novel calcium-independent phospholipase A₂ and its physiological roles in development and immunity of a lepidopteran insect, <i>Spodoptera exigua</i>

Sadekuzzaman, Md.,Gautam, Neelam,Kim, Yonggyun PERGAMON 2017 DEVELOPMENTAL AND COMPARATIVE IMMUNOLOGY Vol. No.

<P><B>Abstract</B></P> <P>Phospholipase A<SUB>2</SUB> (PLA<SUB>2</SUB>) catalyzes hydrolysis of ester linkage at <I>sn-2</I> position of phospholipids. At least 15 groups (I-XV) of PLA<SUB>2</SUB> gene superfamily are associated with various physiological processes such as digestion, secretion, immunity, and maintenance of membrane integrity. This study suggests that various insects encode putative Group VI PLA<SUB>2</SUB>s representing intracellular and calcium-independent PLA<SUB>2</SUB>s (iPLA<SUB>2</SUB>). These insect iPLA<SUB>2</SUB>s are separated into at least two subgroups: iPLA<SUB>2</SUB>A (Group VIA-like) and iPLA<SUB>2</SUB>B (non-Group VIA). Most insects encode genes of iPLA<SUB>2</SUB>B type, although their biological functions are currently unknown. This study predicted a novel iPLA<SUB>2</SUB> from <I>Spodoptera exigua</I> (a lepidopteran insect) (SeiPLA<SUB>2</SUB>B) and analyzed its physiological functions by RNA interference (RNAi). <I>SeiPLA</I> <SUB> <I>2</I> </SUB> <I>B</I> encodes 336 amino acid sequence with a predicted size of about 36.6 kDa and an isoelectric point at pH 8.61. It possesses a lipase catalytic site, but does not have ankyrin repeats in the amino terminal region. Phylogenetic analysis indicated that <I>SeiPLA</I> <SUB> <I>2</I> </SUB> <I>B</I> was clustered with other Group VI iPLA<SUB>2</SUB>s, in which <I>SeiPLA</I> <SUB> <I>2</I> </SUB> <I>B</I> was closely associated with Group VIF gene while <I>SeiPLA</I> <SUB> <I>2</I> </SUB> <I>A</I> was closely related to Group VIA gene. <I>SeiPLA</I> <SUB> <I>2</I> </SUB> <I>B</I> was expressed in all developmental stages of <I>S. exigua</I>. In larval stage, <I>SeiPLA</I> <SUB> <I>2</I> </SUB> <I>B</I> was expressed in fat body, hemocyte, and epidermis, but not in digestive tract. <I>SeiPLA</I> <SUB> <I>2</I> </SUB> <I>B</I> RNAi significantly reduced PLA<SUB>2</SUB> enzyme activities and resulted in developmental retardation and immunosuppression. Though RNAi treatment did not significantly change fatty acid composition in fat body lipids, it significantly increased lipid peroxidation. Taken together, our results suggest that SeiPLA<SUB>2</SUB>B plays important roles in the development and immunity of <I>S. exigua.</I> </P> <P><B>Highlights</B></P> <P> <UL> <LI> A novel intracellular and calcium-independent PLA<SUB>2</SUB> (SeiPLA<SUB>2</SUB>B) gene is predicted. </LI> <LI> Its RNA interference caused oxidative damage by increasing lipid peroxidation. </LI> <LI> SeiPLA<SUB>2</SUB>B plays roles in development and immunity. </LI> </UL> </P>

요추 추간판 탈출증 환자에 있어서 Phospholipase A₂와 통증과의 관계

문성환,박문수,김향,김학선,이환모 대한척추외과학회 2002 대한척추외과학회지 Vol.9 No.1

Association of Single Nucleotide Polymorphisms in the Prostaglandin-endoperoxide Synthase 2 (PTGS2) and Phospholipase A₂ Group IIA (PLA2G2A) Genes with Susceptibility to Esophageal Squamous Cell Carcinoma

Liu, Fen,Wei, Wen-Qiang,Cormier, Robert T.,Zhang, Shu-Tian,Qiao, You-Lin,Li, Xin-Qing,Zhu, Sheng-Tao,Zhai, Yan-Chun,Peng, Xiao-Xia,Yan, Yu-Xiang,Wu, Li-Juan,He, Dian,He, Yan Asian Pacific Journal of Cancer Prevention 2014 Asian Pacific journal of cancer prevention Vol.15 No.4

Background: The prostaglandin-endoperoxide synthase 2 (PTGS2) and phospholipase A2 group IIA (PLA2G2A) genes encode enzymes that are involved in arachidonic acid and prostaglandin biosynthesis. Dysregulation of both genes is associated with inflammation and carcinogenesis, including esophageal squamous cell carcinoma (ESCC). We therefore hypothesized that there is an association between single nucleotide polymorphisms (SNPs) in these genes and susceptibility to ESCC. Methods: We performed a gene-wide tag SNP-based association study to examine the association of SNPs in PTGS2 and PLA2G2A with ESCC in 269 patients and 269 healthy controls from Taihangshan Mountain, Henan and Hebei Provinces, the rural area of China which has the highest incidence of esophageal cancer in the world. Thirteen tag SNPs in PLA2G2A and 4 functional SNPs in PTGS2 were selected and genotyped using a high-throughput Mass Array genotyping platform. Results: We found a modest increased risk of ESCC in subjects with the PTGS2 rs12042763 AA genotype (OR=1.23; 95% CI, 1.00-3.04) compared with genotype GG. For PLA2G2A, a decreased risk of ESCC was observed in subjects with the rs11677 CT (OR=0.51, 95%CI, 0.29-0.85) or TT genotype (OR=0.51, 95%CI, 0.17-0.96) or the T carriers (CT+TT) (OR=0.52, 95%CI, 0.31-0.85) when compared with the CC genotype. Also for PLA2G2A, rs2236771 C allele carriers were more frequent in the control group (P=0.02). Subjects with the GC (OR=0.55, 95%CI, 0.33-0.93) or CC genotype (OR=0.38, 95% CI, 0.16-0.94) or the C carriers (GC+CC) (OR=0.52, 95%CI, 0.32-0.85) showed a negative association with ESCC susceptibility. Conclusions: Our results suggest that PTGS2 and PLA2G2A gene polymorphisms may modify the risk of ESCC development.

C6세포에서 phospholipase A2촬성에 대한 ATP의 작용

심상수(Sang Soo Sim),김명준(Myung June Kim),윤신희(Shin Hee Yoon),김창종(Chang Jong Kim),조양혁(Yang Hyeok Jo) 대한약학회 2001 약학회지 Vol.45 No.4

To investigate action of ATP on ischemia-induced brain injury we measured phospholipase A2 activity and nitric oxide (NO) production in C6 cells. ATP alone did not have any influence on phospholipase A2 activity but increased NO production. Glutamate (1 Mm) significantly increased phospholipase A2 activity whereas did not increased NO production. ATP significantly inhibited phospholipase A2 activation induced by 0.1μM A23187, 1 Mm glutamate and 1 Mm H2O2, but did not inhibited 1mM PMA-induced phospholipase A2 activation. From the above results, it is suggested that the action of ATP in C6 cells has dual actions, such as the inhibition of agonist-induced phospholipase A2 activation and the increase of NO production.

Overexpression of Phospholipase A₂ Group IIA in Esophageal Squamous Cell Carcinoma and Association with Cyclooxygenase-2 Expression

Zhai, Yan-Chun,Dong, Bin,Wei, Wen-Qiang,He, Yan,Li, Xin-Qing,Cormier, Robert T.,Wang, Wei,Liu, Fen Asian Pacific Journal of Cancer Prevention 2014 Asian Pacific journal of cancer prevention Vol.15 No.21

Background: Esophageal cancer is one of the most frequently occurring malignancies and the seventh leading cause of cancer-related deaths in the world. The esophageal squamous cell carcinoma (ESCC) is the most common histological type of esophageal cancer worldwide. Materials and Methods: Our goal in this study was to detect phospholipase A2 Group IIA (PLA2G2A) and cyclooxygenase-2 (COX-2) immuno-expression in ESCC in a high-risk population in China. Results: Positive expression of PLA2G2A protein was observed in 57.2% (166/290) of the cases, while COX-2 was found in 257 of 290 samples (88.6%), both PLA2G2A and COX-2 being expressed in 153 cases (52.8%), with a significant agreement (Kappa=0.091, p=0.031).Overexpression of PLA2G2A was significantly correlated with the depth of invasion (p=0.001). Co-expression of PLA2G2A and COX-2 not only significantly correlated with the depth of invasion (p=0.004) but also with TNM stage (p=0.04). Conclusions: Our results showed that in patients with ESCC, PLA2G2A overexpression and PLA2G2A co-expression with COX-2 is significantly correlated with advanced stage. The biological role and pathophysiologic regulation of PLA2G2A and COX-2 overexpression in ESCC deserve further investigation.