蛇草肾炎颗粒对C-BSA膜性肾病小鼠ERK1/2和cPLA2通路的影响

目的:观察蛇草肾炎颗粒对C-BSA膜性肾病小鼠ERK1/2(细胞外调节蛋白激酶)和cPLA2(细胞质磷脂酶A2)信号通路蛋白的影响,探讨蛇草肾炎颗粒对膜性肾病的作用机理。方法:将50只小鼠随机分为正常对照组、模型组、蛇草肾炎颗粒高剂量组(7.8 g/kg)、低剂量组(3.9 g/kg)及洛汀新组(1.26 g/kg)。采用小鼠尾静脉注射C-BSA复制膜性肾病模型,造模后各组按照小鼠体质量分别灌胃给药;正常组、模型组灌胃等量生理盐水,每天1次,灌胃28 d。观察各组小鼠24 h尿蛋白定量、血清白蛋白的变化情况;末次给药后,采用Western blotting法检测肾组织ERK1/2、p-ERK1/2、cPLA2、p-cPLA2信号通路蛋白。结果:与正常对照组比较,模型对照组小鼠24 h尿蛋白定量显著升高(P<0.01);与模型对照组比较,洛汀新组、蛇草肾炎颗粒低、高剂量组小鼠24 h尿蛋白定量均显著降低(P<0.05或P<0.01)。与正常组比较,模型组小鼠血清白蛋白水平显著降低(P<0.05);与模型组比较,蛇草肾炎颗粒高、低剂量组血清白蛋白含量均显著升高(P<0.01)。与正常对照组比较,模型组小鼠肾组织ERK1/2和cPLA2磷酸化表达显著升高(P<0.01);与模型组比较,蛇草肾炎颗粒高、低剂量组肾组织ERK1/2和cPLA2磷酸化的水平均明显降低(P<0.05、P<0.01)。结论:蛇草肾炎颗粒能够显著降低膜性肾病小鼠尿蛋白,升高血清白蛋白水平,降低ERK1/2和cPLA2磷酸化蛋白表达是其作用机制之一。

大鼠脊髓损伤后GFAP、cPLA2的表达变化及相关性研究

目的研究大鼠脊髓半横断损伤后胶质纤维酸性蛋白(glial fibrillary acidic protein,GFAP)和胞浆型磷脂酶A2(cytosolic phospholipase A2,cPLA2)的表达与分布,并分析其相关性,探讨两者在脊髓损伤后表达的意义和可能的内在关系。方法 30只SD大鼠随机分为脊髓损伤组(1、3、7、14和21 d)与假手术组,脊髓损伤组用T10节段半横断法(右侧)造模,而假手术组只行椎板切除不损伤脊髓。在各时间点取材后用尼氏染色观察脊髓组织病理学改变,采用免疫组织化学法检测脊髓组织GFAP与cPLA2表达,并经Motic Images Advanced软件处理后进行Pearson相关性分析。结果尼氏染色示脊髓损伤后神经元坏死,肿胀,空泡形成,形态不完整。免疫组织化学法示脊髓损伤后可见GFAP与cPLA2表达均增高,其中GFAP在各时间点表达逐渐增多,cPLA2表达在损伤后14 d达到高峰,GFAP与cPLA2阳性产物面积相关性分析提示其相关系数为0.74,差异无统计学意义(P=0.15),两者平均光密度值行相关分析后,相关系数为0.90,具有统计学意义(P=0.04)。结论大鼠脊髓损伤后GFAP与cPLA2表达均增多,且两者在损伤过程中表达具有相关性。

cPLA2和COX-2在肺癌组织中的表达及生物学意义

目的探讨磷脂酶A2(cPLA2)和环氧合酶(COX-2)在肺癌中的表达及生物学意义,为肺癌的靶向治疗提供参考依据。方法采用RT-PCR检测肺癌及癌旁组织中cPLA2、COX-2 mRNA的表达,同时采用免疫组织化学SP法检测相应标本中cPLA2和COX-2蛋白的表达。结果肺癌组织中cP-LA2和COX-2 mRNA的表达量均明显高于癌旁组织(P<0.05)。cPLA2蛋白在肺癌组织中的阳性表达率为76.7%(46/60),癌旁组织中未见表达。肺癌组织中cPLA2蛋白的阳性表达率与肿瘤大小、组织类型、TNM分期、分化程度和淋巴结转移均无关(P>0.05)。COX-2蛋白在肺癌组织中的阳性表达率为73.3%(44/60),明显高于在相应癌旁组织中的表达[13.3%(8/60)](χ2=21.99,P<0.01)。肺癌组织中COX-2蛋白的阳性表达率与肿瘤大小、组织类型无关(P>0.05);而与淋巴结转移、TNM分期、分化程度有关,随着淋巴结的转移、组织分化程度的降低和临床分期的增加,肺癌组织中COX-2蛋白的阳性表达率逐渐增高,差异有统计学意义(P<0.05)。结论 COX-2和cPLA2的高表达可能在肺癌的发生发展起着重要作用,它们可能为肺癌的早期诊断和开发肺癌的靶向治疗提供一定的临床依据。

高盐饮食增强内皮细胞TRPV4与cPLA2的空间偶联作用

目的探究内皮细胞中香草素受体4型瞬时感受器电位通道(TRPV4通道)与胞浆型磷脂酶A2(c PLA2)在空间上的偶联作用。方法通过设定梯度盐(培养基中外源加入20、40、80、160 mol·L^(-1)Na Cl)确定高盐处理内皮细胞的最适盐浓度;在细胞水平上,通过免疫荧光能量共振转移(immuno-FRET)技术检测人微血管内皮细胞(HMEC)和小鼠胸主动脉原代内皮细胞中TRPV4与c PLA2的空间偶联作用;在动物组织水平,通过同样的技术检测正常小鼠与高盐诱导的高血压小鼠的胸主动脉血管环上的内皮细胞中TRPV4与c PLA2的空间偶联情况。结果高盐诱导内皮细胞的最适盐浓度为40 mol·L^(-1)Na Cl;在细胞水平和动物组织水平上,高盐处理的HMEC、小鼠胸主动脉原代内皮细胞和高盐诱导的高血压小鼠胸主动脉血管环上的内皮细胞中,TRPV4与c PLA2的空间偶联作用均明显增强。结论在细胞和组织水平上,高盐饮食能增强内皮细胞中TRPV4与c PLA2的空间偶联作用,这种空间偶联可能进一步引起两者的功能偶联,为血管内皮功能紊乱的研究提供了新的思路。

L-谷氨酰胺通过p38 MAPK/cPLA2信号通路抑制变应性接触性皮炎

[目的]探讨L-谷氨酰胺(Gln)对变应性接触性皮炎(ACD)模型小鼠的影响及其机制.[方法]利用2,4-二硝基氟苯反复刺激小鼠皮肤建立ACD模型,观察Gln对ACD模型小鼠耳肿胀度、组织病理改变、耳组织p38丝裂原活化蛋白激酶(MAPK)及胞浆型磷酯酶A2(cPLA2)蛋白表达和血清肿瘤坏死因子-α(TNF-α)及白细胞介素4(IL-4)等细胞因子表达的影响.[结果 ]与正常对照组比较,Gln显著抑制ACD模型小鼠耳肿胀度(P<0.05),明显降低皮肤组织p38 MAPK,cPLA2蛋白及血清TNF-α,IL-4炎性细胞因子表达水平(P<0.05).[结论]Gln治疗小鼠ACD有效,机制认为与p38 MAPK/cPLA2信号通路及TNF-α,IL-4炎性细胞因子表达有关系.

参芪蛭龙汤通过ERK/cPLA2信号通路对膜性肾病小鼠的治疗作用及机制研究

目的:观察参芪蛭龙汤方对膜性肾病(MN)小鼠的治疗作用及可能的机制。方法:通过多次尾静脉注射阳离子牛血清白蛋白制备MN模型小鼠。造模结束后,依据尿蛋白数值将MN小鼠随机分为模型对照组(等量蒸馏水)、参芪蛭龙汤低剂量组(12 g生药/kg)、参芪蛭龙汤高剂量组(24 g生药/kg)和雷公藤多苷片组(14 mg/kg)。另取10只正常小鼠作为空白对照组(等量蒸馏水)。连续给药4周,每天灌胃给药1次。给药结束后,收集24 h尿液,测定尿蛋白数值;眼球取血,测定肾功、肝功、血脂及血清IL-6、IL-4、TNF-α含量;HE染色观察肾脏病理变化;免疫组化染色观察肾脏IgG沉积情况;Western-blot法测定肾脏组织ERK1/2和cPLA2蛋白表达;RT-PCR法检测肾脏组织Neph-1、Nephrin及Podocin基因表达。结果:与模型组比较,参芪蛭龙汤组病理变化明显改善;参芪蛭龙汤低、高剂量组可显著降低MN小鼠尿蛋白数值;降低血脂TC及TG水平(P<0.05或P<0.01);升高ALB及TP水平(P<0.05或P<0.01);对CRE、UREA及UA水平无显著影响(P>0.05);降低血清IL-6、IL-4及TNF-α含量(P<0.05或P<0.01);显著下调MN小鼠肾脏组织中p-ERK1/2和p-cPLA2蛋白表达水平(P<0.05或P<0.01);显著升高肾脏组织Neph1、Nephrin及Podocin mRNA基因表达水平(P<0.01)。结论:参芪蛭龙汤对MN小鼠具有较好的治疗作用,作用机制与调节Nephrin-podocin-Neph1受体复合物相关基因表达,保护肾小球滤过屏障;抑制ERK/cPLA2途径的激活,减轻肾小球上皮细胞损伤,从而减少促炎细胞因子分泌有关。

UCN enhances TGF-β-mediated mitoinhibition of VSMCs via counteracting TGF-β-induced cPLA2 expression and activation

OBJECTIVE Urocortins(UCNs)and transforming growth factor-β(TGF-β)have been demonstrated to participate in various cardiovascular diseases,many of which involve VSMCs proliferation.And cytosolic phospholipase A2(c PLA2)-mediated arachidonic acid(AA)release is an important cause of vascular smooth muscle cells(VSMCs)proliferation.The work was to investigate the regulation of VSMCs proliferation by UCN/TGF-βand whether c PLA2 was a link between their signaling pathways.METHODS VSMC proliferation was measured by MTT assay and immunofluorescence microscopy.Using cell flow cytometry,the changes in the cell cycle phases were investigated.si RNA was used to knockdown Smad2 and smad3 genes.Lentiviral Vector Particle was performed to over express c PLA2 gene.RESULTS Both UCN and TGF-βinhibited VSMCs proliferation and an additive effect was observed when the cells were treated with UCN plus TGF-β.TGF-βincreased the percentage of cells in G1-phase while UCN increased the cell percentage in G2-phase with a concomitant decrease in S-phase.Neither knockdown of smad2 nor smad3 reversed the role of TGF-β.Furthermore,c PLA2expression was increased by TGF-βbut decreased by UCN and UCN attenuated TGF-β-induced c PLA2 expression.In primary VSMCs,TGF-βinduced c PLA2 phosphorylation,and this effect was also attenuated by UCN.Similar to UCN,the c PLA2 inhibitor,pyrrophenone(PYR),also played a role in enhancing TGF-β-mediated mitoinhibition.Inversely,over-expression of c PLA2 eliminated the effect of UCN on the mitoinhibition.CONCLUSION The pretreatment with UCN counteracted TGF-β-mediated c PLA2 expression and activation,thereby contributing to TGF-β-mediated mitoinhibition of VSMCs.

Therapeutic effect and mechanism of Shenqi Zhilong Decoction on mice with membranous nephropathy through ERK/cPLA2 signaling pathway

Objective: To explore the therapeutic effect and underlying mechanism of Shenqi Zhilong Decoction on mice with membranous nephropathy (MN). Methods:Mice with MN was established by injecting cationic bovine serum albumin (c-BSA) into tail vein for several times. model mice were randomly divided into MN group (equal amount of distilled water), Shenqi Zhilong Decoction low dose group (12 g crude drug/kg), Shenqi Zhilong Decoction high dose group (24 g crude drug/kg), and Tripterygium wilfordii polyglycoside tablet group (14 mg/ kg). Another 10 un-treatment mice were taken as control group (equal amount of distilled water). The drug was administered orally once a day for 4 weeks. After the last administration, 24 hours urine was collected to determine the urinary protein content;blood from inner canthus was collected to measure the changes of kidney function, liver function, blood lipid and levels of IL-6, IL-4 and TNF-α in serum in each group;HE staining was used to observe the pathological changes of kidney. Immunohistochemical staining was used to observe the expression of IgG in kidney. The protein expression of ERK1/2 and cPLA2 in renal tissues was determined by Western-blot method. The gene expression of Neph1, Nephrin and Podocin mRNA in kidney tissues were detected by RT-PCR. Results: Compared with model group, Shenqi Zhilong decoction at low-dose and high-dose could significantly reduce the value of urine protein in MN mice;Decreased TC and TG levels (P<0.05 or P<0.01);Increased the levels of ALB and TP in liver function (P<0.05 or P<0.01);has no significant effects on the levels of CRE, UREA and UA in renal function (P>0.05). Decreased the contents of IL-6, IL-4 and TNF-α in serum (P<0.05 or P<0.01);Significantly down-regulated the protein expression levels of p-ERK1/2 and p-cPLA2 in kidney tissues of MN mice (P<0.05 or P<0.01);Significantly increased the expression levels of NephP1, Nephrin and Podocin mRNA in renal tissues (P<0.01). Conclusion: Shenqi Zhulong Decoction has a good therapeutic effect on MN mice, and the mechanism of action is related to regulate the expression of related genes of Nephrin-Podocin-Neph1 receptor complex for protecting the glomerular filtration barrier, and inhibite the activation of ERK/cPLA2 pathway for relieving damage of GEC and reduceing secretion of pro-inflammatory cytokines.

The Novel cPLA2 Inhibitor AK106-001616 Has a Protective Effect on SOD1G93A-Induced Cell Death in NSC34 Murine Motor Neuron-Like Cell

The expression of cytosolic phospholipase A2 (cPLA2) expression is up-regulated in animal model of ALS and in patients with familial amyotrophic lateral sclerosis (fALS). Inhibition of cyclooxygenase 2 (COX2), which is a downstream enzyme of cPLA2, ameliorates the impairment of motor function in the ALS model mice. Therefore, the arachidonic acid cascade, including the cPLA2-COX2 pathway, is an important therapeutic target of ALS. The current study was designed to investigate the potential of AK106-001616, an inhibitor of cPLA2, in protection of motor neuron cell death induced by mutant superoxide dismutase (SOD1G93A). AK106-001616 (1 - 10 μM) protected NSC34 cells (mouse motor neuron like cells) against SOD1G93A-induced motor neuron cell death. Furthermore, aspirin, an inhibitor of COX1/2, reduced the SOD1G93A-induced motor neuron cell death at a concentration that inhibited COX2. Celecoxib, a selective COX2 inhibitor, also reduced the SOD1G93A-induced motor neuron cell death. These results suggest that the arachidonic acid cascade is important for SOD1G93A-induced motor neuron cell death and AK106-001616 has a potent neuroprotective effect against it. AK106-001616 may be a useful therapeutic agent against SOD1G93A-induced ALS.

单纯性胰腺创伤后大鼠胰腺磷脂酶A2活性及其亚型表达特点

目的构建大鼠单纯性胰腺创伤模型并观察大鼠血清与胰腺组织中磷脂酶A2(PLA2)活性及其亚型表达特点。方法将120只Wistar大鼠按1:2随机分为对照组及撞击组,并分别分为6h、24h、72h、7d 4个亚组,对照组每个时间点10只,撞击组每个时间点20只。各撞击组采用BIM-Ⅲ生物撞击机构建胰腺创伤大鼠模型;对照组仅行开腹手术,翻动胰腺数次后关腹。检测各组大鼠血清与胰腺组织PLA2活性及PLA2各亚型在胰腺组织中的表达与分布特点。结果撞击组大鼠血清与胰腺组织中PLA2活性在创伤后6h开始升高,血清PLA2于72h达高峰,胰腺组织PLA2于24h达高峰,7d时血清与胰腺组织PLA2活性仍高于对照组(P<0.05)。胰腺组织中分泌型磷脂酶A2(sPLA2)在RNA水平及蛋白水平表达均高于胞质型磷脂酶A2(cPLA2)与钙非依赖型磷脂酶A2(iPLA2)。结论胰腺创伤后,血清与胰腺组织PLA2活性均显著增高,PLA2各亚型中,sPLA2在胰腺表达增强最为显著。胰腺创伤后6h^7d均可检出血PLA2活性,有助于胰腺创伤的辅助诊断。

转染反义磷脂酶A_2寡核苷酸抑制LPS诱导Hela细胞分泌IL-1β和IL-6

目的 :研究胞浆型磷脂酶A2 (cytosolicphospholipaseA2 ,cPLA2 ,85 kDPLA2 )在脂多糖诱导Hela细胞释放细胞因子IL 1β、IL 6的信号机制中的作用。 方法 :①利用脂质体将cPLA2 的反义寡核苷酸转染入Hela细胞 ,通过聚合酶链式反应(RT PCR)及Western杂交印迹分析检验胞浆中cPLA2 水平的变化 ;②用放免方法检测不同时间培养上清中的IL 1β、IL 6浓度。结果 :①转染后cPLA2 的蛋白表达明显减少 ,但mRNA水平未见显著改变 ;②Hela细胞释放IL 1β、IL 6随cPLA2 反义寡核苷酸浓度的增加而减少。结论 :cPLA2 在LPS刺激的Hela细胞释放炎性细胞因子IL 1β、IL 6的过程中可能发挥重要的信号传递作用。

胞内型磷脂酶A2基因亚克隆与基因表达

目的 研究花生四稀酸在细胞中的释放及前列腺素的合成，构建与花生四稀酸释放相关的表达系统。即胞内型磷脂酶Ａ２（ｃｙｔｏｓｏｌｉｃ ｐｈｏｓｐｈｏｌｉｐａｓｅ ｃＰＬＡ２）ｃＤＮＡ－ｐＲＣ／ＣＭＶ。方法 从克隆载体ｐＭＴ２用Ｓａｌ１制备ｃＰＡＬ２ｃＤＮＡ；平末端连接ｃＰＡＬ２－ｐＲＣ／ＣＭＶ；转染鼠巨噬细胞及筛选正向阳性克隆；ｎｏｒｔｈｅｒｎ ｂｌｏｔ检测ｃＰＬＡ２ ｃＤＮＡ基因表达。结果

胞液型磷脂酶A_2cDNA与载体PRCCMV连接及表达

目的 选择适当的表达载体 ,启动胞液型磷脂酶A2 (cytosolicphospholipaseA2 ,简称cPLA2 )cDNA的表达 ,为进一步研究各种因素对该酶活性的影响奠定基础。方法 从cPLA2 PMT2 重组体中获得人cPLA2cDNA片段 ,使之与酶切线性化的PRC CMV载体平末端连接 ,并转染鼠巨噬细胞 ,筛选、扩增阳性细胞 ,提取RNA ,Northern杂交法检测cPLA2 mRNA。结果 应用cPLA2 PRC CMV重组体转染鼠巨噬细胞后 ,经过North ern杂交检测到cPLA2 cDNA的表达。结论 cPLA2 PRC CMV是一有效的表达系统。

Role of Rac1/p38 and ERK-Dependent Cytosolic Phospholipase A2 Activation in Porphyromonas gingivalis-Evoked Induction in Matrix Metalloproteinase-9 (MMP-9) Release by Salivary Gland Cells

Matrix metalloproteinase-9 (MMP-9) is a highly glycosylated endopeptidase implicated in a wide rage of oral mucosal inflammatory and neoplastic diseases, including chronic periodontitis, a persistent mucosal inflammation attributed primarily to infection by oral anaerobe, P. gingivalis. In this study, we explored the role of Rac1 and mitogen-activated protein kinases (MAPKs) in the processes of MMP-9 release in sublingual salivary gland cells exposed to P. gingivalis key endotoxin, cell wall lipopolysaccharide (LPS). We demonstrate that the LPS-elicited induction in the acinar cell MMP-9 release is associated with MAPK, ERK and p38 activation, and occurs with the involvement of Rac1 and cytosolic phospholipase A2 (cPLA2). Further, we reveal that the LPS-induced MMP-9 release involves ERK-mediated phosphorylation of cPLA2 on Ser505 that is essential for its membrane translocation with Rac1, and that this process requires p38 activation. Moreover, we show that phosphorylation and membrane localization of p38 with Rac1-GTP play a pivotal role in cPLA2-dependent induction in MMP-9 release. Thus collectively, our findings infer that P. gingivalis LPS-induced up-regulation in the acinar cell MMP-9 release requires ERK-dependent recruitment of cPLA2 to the membrane localized Rac1/p38 complex.

ELISA测定培养细胞中胞液型磷脂酶A_2的含量

目的建立胞液型磷脂酶A2（cytosolicphospholipaseA2,cPLA2）的ELISA。方法以人cPLA2抗体包被微孔板后，加入抗原（标本），再加入辣根过氧化物酶标记的羊抗人IgG作为第二抗体，经邻苯二胺（OPD）显色后测定其490nm处A值，与标准品对比可得cPLA2含量。结果检测灵敏度可达25．0ng／ml，批内和批间变异系数为5．5％和7．8％，cPLA2含量为56和120mg／ml标本的平均回收率均大于80％。结论该方法是一种测定cPLA2含量的简便可靠方法。

Cytosolic phospholipase A2 S-nitrosylation in ghrelin protection against detrimental effect of ethanol cytotoxicity on gastric mucin synthesis ——Ghrelin in gastric mucosal protection

Ghrelin, a peptide hormone produced mainly in the stomach, has emerged recently as an important regulator of nitric oxide synthase (NOS) and cyclooxygenase (COX) enzyme systems, the products of which play direct cytoprotective function in the maintenance of gastric mucosal integrity. In this study, using gastric mucosal cells, we report on the role of ghrelin in countering the cytotoxic effect of ethanol on mucin synthesis. We show that the countering effect of ghrelin on mucin synthesis was associated with the increase in NO and PGE2 production, and characterized by a marked up-regulation in cytosolic phospholipase A2 (cPLA2) activity. The ghrelin-induced up-regulation in mucin synthesis, like that of cPLA2 activity, was subject to suppression by Src inhibitor, PP2 and ERK inhibitor, PD98059, as well as ascorbate. Moreover, the loss in countering effect of ghrelin on the ethanol cytotoxicity and mucin synthesis was attained with cNOS inhibitor, L-NAME as well as COX-1 inhibitor SC-560. The effect of L-NAME was reflected in the inhibition of ghrelin-induced mucosal cell capacity for NO production, cPLA2 S-nitrosylation and PGE2 generation, whereas COX-1 inhibitor caused only the inhibition in PGE2 generation. Our findings suggest that the activation of gastric mucosal cPLA2 through cNOS-induced S-nitrosylation plays an essential role in the countering effect of ghrelin on the disturbances in gastric mucin synthesis caused by ethanol cytotoxicity.

乳腺癌转移中的磷脂酰胆碱和溶血磷脂酰胆碱分析（英文）

我们以前曾报道花生四烯酸(arachidonic acid,AA)代谢产物可以促进乳腺癌细胞增殖和迁移.为了进一步寻找维持高转移乳腺癌细胞中AA高水平代谢的内源机制,深入探求AA代谢促进乳腺癌细胞转移的分子机理,我们应用HPLC/ESI/MSn技术检测和分析了乳腺癌MCF-7和高转移乳腺癌LM-MCF-7细胞中溶血磷脂酰胆碱(lysophosphatidylcholines,Lyso PCs)和磷脂酰胆碱(phosphatidylcholines,PCs)的成分和含量.发现了10种Lyso PC的含量在LM-MCF-7细胞中显著高于MCF-7细胞,有6种PC可水解产生AA,它们在LM-MCF-7细胞中的含量显著低于MCF-7细胞,提示这些溶血磷脂含量的升高和磷脂含量的降低可能与乳腺癌转移相关.在LM-MCF-7细胞中,COX-2抑制剂吲哚美辛(indomethacin,Indo)和LOX抑制剂(nordihydroguaiaretic acid,NDGA)共同作用可明显下调c PLA2的活性,应用HPLC-ESI-MSn技术比较c PLA2活性下调前后LM-MCF-7细胞中Lyso PC和PC含量的变化,发现其中4种PC可被c PLA2水解产生AA.还发现,细胞内Lyso PC与PC的比值可以反映c PLA2的活性.通过以上研究进一步证实了由c PLA2活性调节的AA释放及代谢对乳腺癌转移具有重要作用.

地塞米松诱发小鼠腭裂时AnnexinⅠ、cPLA_2和PCNA表达的研究

目的 检测膜联蛋白 (AnnexinⅠ )、胞质磷脂酶A2 (cytosolicphospholipaseA2 ,cPLA2 )、增殖细胞核抗原 (proliferatingcellmuclearantigen ,PCNA)在地塞米松诱导小鼠腭裂腭胚突中的表达变化情况。探讨腭裂的发生机制和预防方法。方法 A/J和C5 7BL/ 6 j受孕小鼠随机分为正常对照组、地塞米松致畸组 (DEX组 )、VitB12 致畸拮抗组 (DE +Vit组 )和VitB12 组。应用蛋白质免疫印迹的方法检测AnnexinⅠ、cPLA2 和PCNA的相对含量。结果 A/J小鼠中 ,在正常对照组中 ,随着腭胚突的发育 ,AnnexinⅠ表达逐渐升高 ,其余各实验组也呈相似变化。在DEX组和DEX +VitB12 组中 ,AnnexinⅠ的表达明显较对照组和VitB12 组高 ,对照组与VitB12 组间差异无显著性。而cPLA2 则和PCNA呈相反表达趋势 ,在正常小鼠腭胚突中表达随小鼠发育逐渐下降。在C5 7BL/ 6 j小鼠中 ,正常对照组与DEX组间在AnnexinⅠ和cPLA2 测量差异无显著性。结论 AnnexinⅠ和cPLA2 介导了糖皮质激素诱发腭裂的过程。VitB12 未能阻遇DEX上调AnnexinⅠ的表达 ,但可以对抗DEX抑制cPLA2 的作用 ,这可能是VitB12

大鼠脑缺血再灌注后海马区cPLA_2、Caspase-3的分布

目的 研究脑缺血再灌注后cPLA2 和Caspase 3在海马区的分布变化。方法 使用大鼠局灶性脑缺血再灌注模型 ,采用免疫组化方法 (SABC法 )。结果 脑缺血 2h再灌注 2h后即有cPLA2 和Caspase 3阳性表达 ;再灌注 2 4h后表达达到高峰。结论 大鼠脑缺血再灌后后 ,cPLA2 在损伤过程中起到非常重要的作用 ,同时Caspase 3水解cPLA2 ,对cPLA2 酶活性起到调节作用 ,它们共同参与海马区神经元的损伤。