P38MAPK和P13K/Akt信号通路在大鼠糖尿病神经病理性疼痛中的交互作用

目的 观察p38MAPK和P13K/Akt信号通路在大鼠糖尿病神经病珲性疼痛中的交互作用.方法 Wistar大鼠腹腔单次注射链脲菌素65 mg/kg制作糖尿病神经病理痛模型.4周后用von frey纤维测双后足机械痛阈,痛阈明显下降为糖尿病神经病理性疼痛造模成功.将96只成模大鼠随机均分为三组:糖尿病神经病理痛组(D组),PI3K抑制药组(E组)和p38MAPK抑制药组(F组).另取同窝大鼠32只作为对照组(C组).成模后的每周周一,E组和F组大鼠分别静脉注射P13K抑制药Wortmannin 0.5 mg/kg和p38MAPK抑制药SB203580 1 mg/kg,直至处死大鼠.于给药前(T1)和给药后第2周末(T2)、第4周末(T3)、第6周末(T4)分别随机取8只大鼠,检测机械缩足反应阈值(MWT)、神经传导速度(NCV)、脊髓和背根神经节(DRG)磷酸化Akt(p-Akt)水平和磷酸化p38MAPK(p-p38MAPK)水平.结果 与C组比较,D、E和F组在T1～T4时MWT下降,NCV减慢,p-Akt和p-p38MAPK水平升高(P<0.05);与D组比较,E和F组在T2～T4时MWT升高,NCV增快,E和F组p-Akt明显下降,F组p-p38MAPK水平明显下降(P<0.05).结论 P38MAPK通过激活其下游的P13K/Akt信号通路参与了糖尿病大鼠神经病理痛的形成和维持.     

P38MAPK与糖尿病肾病

有丝分裂原蛋白激酶(MAPK)是生物体内重要的信号转导系统之一。P38MAPK属于MAPK的一个亚族,是广泛存在于细胞浆内的含有丝氨酸/苏氨酸残基的蛋白质激酶。它在糖尿病肾病(DN)的形成中起着非常重要的作用,糖尿病状态下多种因素的作用激活P38MAPK,促进肾脏细胞肥大,ECM积聚,导致肾小球硬化和肾小管间质纤维化。故深入研究P38MAPK在DN发病中的作用,有助于阐述DN的发病机理,为开发防治DN的新药提供科研依据。     

p38丝裂原活化蛋白激酶在糖尿病大鼠神经病理性痛中的作用

目的探讨p38丝裂原活化蛋白激酶（p38MAPK）在链脲菌素诱导的糖尿病大鼠神经病理性痛中的作用。方法雌性Wistar大鼠31只，3月龄，体重180～220g，随机分为3组：对照组（C组，n=10）、糖尿病神经病理性痛组（D组，n=11）和p38MAPK抑制剂组（Ⅰ组，n=10）。D组、Ⅰ组单次腹腔注射链脲菌素65mg／kg制备糖尿病模型。糖尿病模型制备成功后，Ⅰ组尾静脉注射p38MAPK抑制剂SB203580 0．5mg／kg，1次／周，连续4周；C组和D组尾静脉注射等体积的生理盐水。给药4周后，测定机械缩足反应阈值（MWT）、左侧坐骨神经传导速率（NCV）、背根神经节（DRG）和脊髓的磷酸化p38MAPK水平。结果与C组比较，D组、Ⅰ组MWT下降，NCV减慢，伴有脱髓鞘现象，DRG和脊髓的磷酸化p38MAPK水平升高；与D组比较，Ⅰ组MWT升高，NCV增快，脱髓鞘程度减轻，DRG和脊髓的磷酸化p38MAPK水平下降。结论p38MAPK信号转导通路参与了糖尿病大鼠神经病理性痛的形成。     

ERK and p38 MAP kinase are required for rat renal development

BACKGROUND: We previously demonstrated that extracellular signal-regulated protein kinase (ERK) and p38 mitogen-activated protein (MAP) kinase (p38) are strongly expressed in the embryonic kidney. In the present study, we investigated the role of ERK and p38 during kidney development. METHODS: Rat metanephroi were cultured from 15-day-old embryos, and exposed to inhibitors of MEK, an activator of ERK, PD98059 (300 micromol/L), U0126 (10 micromol/L), or a p38 inhibitor SB203580 (30 micromol/L) 24 to 120 hours after the start of culture. Growth of metanephroi was measured by surface area and thymidine incorporation. Ureteric buds and glomeruli were identified by labeling with Dolichos biflorus lectin and peanut agglutinin, respectively. PCNA staining and TUNEL assay were performed on kidney sections. The level of apoptosis was evaluated by examining DNA ladder formation. RESULTS: Growth of metanephroi was significantly inhibited by SB203580 but not by PD98059 or U0126. Ureteric bud branching was not affected by SB203580 or MEK inhibitors. Glomerular number was markedly reduced by SB203580 and to a lesser extent by U0126 (14 +/- 2 and 48 +/- 10% of controls, respectively). On histological examination, the number of tubuloglomerular structures was reduced in MEK inhibitor-treated metanephroi compared to controls. Very few mesenchymal condensates were observed in kidneys incubated with SB203580. PCNA-positive cells were reduced in SB203580-treated metanephroi compared to control and PD98059-treated kidneys. Apoptosis was increased in SB203580-treated kidneys and to a lesser extent in PD98059-treated cultures. CONCLUSIONS: Both ERK and p38 are required for renal development. ERK appears to play a role in nephrogenesis and p38 for kidney growth and nephrogenesis.     

甲状腺乳头状癌中p16/p53/p38MAPK/Wip1通路的改变及意义

目的研究甲状腺乳头状癌中p16/p53/p38MAPK/Wip1通路的改变及临床意义。方法应用免疫组织化学方法,检测70例PTC组织和20例正常甲状腺组织中Wip1,p38MAPK,p53及p16蛋白的表达,并将Wip1蛋白高表达与p38MAPK,p53,p16蛋白表达进行相关性分析。结果 PTC组织中Wip1蛋白高表达率为64.3%(45/70),与正常甲状腺组织(0/20)有明显差异(P<0.01);Wip1高表达率在年龄、性别、肿瘤大小、淋巴结转移、病理分期各因素分组间差异无统计学意义(均P>0.05);Wip1蛋白高表达与p3 8 MAPK,p5 3,p1 6蛋白表达呈负相关(r=-0.6 2 0,r=-0.3 5 6,r=-0.550,均P<0.0 1)。结论PTC中存在p1 6/p5 3/p3 8 MAPK/Wip1通路异常,其机制可能与Wip1异常上调后抑制p3 8 MAPK,p5 3,p1 6有关。Wip1可能为甲状腺癌治疗的潜在靶点。     

丝裂素活化蛋白激酶p38在前列腺上皮内瘤的表达及其意义

目的：探讨丝裂素活化蛋白激酶p38（p38MAPK,p38）在前列腺肿瘤形成过程中的作用。方法：采用免疫组织化学SP法检测12例前列腺上皮内瘤（PIN）和12例前列腺增生（BPH）组织中p38的表达。结果：p38在PIN中主要分布于腺上皮和肿瘤细胞,呈片状、弥漫状分布。在BPH中主要分布于腺上皮基底细胞,分泌细胞有少量着色。PIN和BPH组织中p38的活性表达分别为（63．83±18．34）％和（48．75±16．94）％,两组间比较,差异有统计学意义（P〈0．05）。结论：PIN组织中p38的活性显著高于BPH。p38活性的增强参与了前列腺上皮的恶性转化,可能在前列腺肿瘤的发生、发展中起着重要作用。     

The phosphatidylinositol 3-kinase, p38, and extracellular signal-regulated kinase pathways are involved in osteoclast differentiation.

Phosphatidylinositol 3-kinase (PI 3-kinase) and mitogen-activated protein kinases (MAPKs) have been implicated in diverse cellular functions, including proliferation, migration, and survival. In this study, we examined the involvement of these kinases in osteoclast differentiation by employing specific inhibitors of the kinases. The osteoclast differentiation was assessed in three different culture systems: a coculture of mouse bone marrow cells with mouse calvarial osteoblasts, a mouse bone marrow cell culture in the presence of receptor activator of NF-kappaB ligand (RANKL) and macrophage-colony stimulating factor (M-CSF), and a culture of bone-resident osteoclast precursor cells driven by RANKL and M-CSF. LY294002, a specific inhibitor of PI 3-kinase, potently inhibited osteoclast differentiation in all culture systems when assessed by both tartrate-resistant acid phosphatase (TRAP) staining and dentine resorption assays. Inhibition of p38 MAPK by SB202190 resulted in a strong suppression in the exogenous RANKL dependent mouse bone marrow and bone resident precursor cell cultures. Another MAPK pathway inhibitor (PD98059), which blocks the activation of extracellular signal-regulated kinase (ERK) by inhibiting the upstream kinase MAPK-ERK kinase (MEK) 1, exerted an inhibitory effect on osteoclast differentiation only at the highest concentration tested (30 micromol/L) in many cases. Whether the signaling pathways involving these kinases are activated by RANKL was also examined. The RANKL-stimulated phosphorylation of Akt, a downstream target of PI 3-kinase, and that of ERK were observed. RANKL also stimulated the activity of p38. These results suggest that PI 3 kinase, p38, and ERK play roles in osteoclast differentiation, at least in part, by participating in RANKL signaling.     

p16/p38 MAPK/p53/Wip1通路在乳腺癌发生、发展中的作用及其临床意义

目的 探讨p16/p38 MAPK/p53/Wip1通路在乳腺癌发生、发展中的作用及其临床意义.方法 应用免疫组织化学方法检测70例乳腺癌组织、癌旁组织、20例正常乳腺组织中Wip1、p53、p38 MAPK、p16蛋白的表达,并对Wip1蛋白高表达与p53、p38、p16蛋白表达进行相关分析.结果 3种组织中Wip1蛋白高表达率分别为62.9%(44/70)、2.9%(2/70)、0(0/20).乳腺癌组织比癌旁组织、正常乳腺组织明显升高(P＜0.01).Wip1蛋白高表达与p53、p38、p16蛋白表达呈负相关(P＜0.01,等级相关系数rs分别为-0.529、-0.626、-0.499).结论 p16/p38 MAPK/p53/Wip1是负反馈通路,它可能在乳腺癌发生发展中起重要作用.     

白细胞介素1β通过p38信号通路上调大鼠肾小球系膜细胞骨调素mRNA的表达

目的 探讨p38信号通路(1938MAPK)在白细胞介素1(IL-1)β介导的大鼠肾小球系膜细胞表达骨调素(OPN)中的作用。方法 应用Western印迹检测p38MAPK在IL-1β诱导的肾小球系膜细胞炎症反应中的活化程度。应用逆转录-聚合酶链式反应(RT-PCR)法观察p38MAPK特异性阻断剂SB203580对IL-1β诱导的系膜细胞促炎症介质OPNmRNA的影响。结果 IL-1β以时间和剂量依赖方式刺激系膜细胞引起p38MAPK的活化，并明显上调系膜细胞OPNmRNA的表达。p38MAPK特异性抑制剂SB203580以剂量依赖性方式显著抑制IL-1β诱导的OPNmRNA的表达。结论 p38MAPK在IL-16介导的肾小球系膜细胞上调表达黏附分子OPN中起重要作用。     

Activation of p38MAPK signaling cascade in a VSMC injury model: role of p38MAPK inhibitors in limiting VSMC proliferation.

INTRODUCTION: P38 mitogen-activated protein kinase (MAPK) has a crucial role in regulating signaling pathways implicated in the cellular events leading to restenosis. We examine p38MAPK activation in response to vascular cell injury, its biological effects and determine whether selective p38MAPK inhibitors, SB220025/SB203580, decrease vascular smooth muscle cell (VSMC) proliferation. METHODS: Human aortic VSMCs were cultured and wounds made on the monolayers to elicit mitogenic responses and induce p38MAPK activation. P38MAPK inhibitor pretreatment, at varying doses (1-100 microM) and treatment duration was used to block p38MAPK phosphorylation. Cytotoxicity, viability, proliferation and apoptosis were determined and expression of p38MAPK/phospho-p38MAPK was obtained by chemiluminiscent immunoblot analysis. RESULTS: Phosphorylation of p38MAPK depended on injury severity and was inhibited by both p38MAPK inhibitors, but not by SB202474, a specific antagonist of p38MAPK inhibitors. VSMCs treated with p38MAPK inhibitors showed a dose-dependent decrease in viable cell number, apoptosis and proliferation, reversing the deleterious effects of p38MAPK activation comparable to controls (p < 0.05). CONCLUSIONS: This wound injury model activates the p38MAPK-signaling cascade in VSMC and causes cell proliferation that can be abrogated by pre-incubation with p38MAPK selective synthetic inhibitors in a time and dose-dependent manner. SB220025 used here for the first time in VSMC reveals itself to be a stronger p38MAPK inhibitor than SB203580 and being a second generation inhibitor may be the preferred drug for novel therapeutic maneuvers.     

Involvement of Rho and p38 MAPK in endothelin-1-induced expression of PGHS-2 mRNA in osteoblast-like cells.

Prostaglandins (PGs) play an important role in bone remodeling because eicosanoids are local mediators of bone metabolism, which can induce physiological and pathological responses of bone tissue. Biosynthesis of PGs is catalyzed by constitutively expressed PG endoperoxide G/H synthase (PGHS) 1 and by the inducible isoform PGHS-2. In MC3T3-E1 osteoblast-like cells, expression of PGHS-2 was shown by mechanical forces, cytokines, growth factors, and hormones. Recently, endothelin (ET) 1-stimulated PGHS-2 mRNA expression was described, leading to a burst in prostaglandin E2 (PGE2) production. In this study, we investigated ET-1-induced signal transduction pathway(s) involved in the PGHS-2 mRNA production. Time course of PGHS-2 mRNA expression reaching the maximum within 45 minutes is in good agreement with the concept of an immediate early gene product. Inhibition of phospholipase C (PLC), phospholipase D (PLD), phosphatidylinositol-3 kinase (PI-3-kinase), and protein kinase C (PKC) had no influence on PGHS-2 synthesis. Using specific blockers of tyrosine kinases indicated involvement of p38 MAPK but not p42/44 MAPK. By preloading cells with exoenzyme C3, we were able to show requirement of the Rho family of G proteins for p38 MAPK phosphorylation and PGHS-2 mRNA synthesis, whereas pertussis toxin (PTX) and cholera toxin (CTX) had no remarkable effect.     

MLK3-MKK3/6-p38MAPK信号转导通路在大鼠内毒素性急性肺损伤中的作用

目的 探讨MLK3-MKK3/6-p38MAPK信号转导通路在大鼠内毒素性急性肺损伤中的作用.方法 健康成年雄性SD大鼠78只,体重200～250 g,随机分为4组:对照组(C组,n=6)、急性肺损伤组(ALI组,n=24)、MLK3抑制剂K252a组(MK组,n=24)和p38MAPK特异性抑制剂SB203580组(MS组,n=24).ALI组尾静脉注射内毒素5 mg/kg制备大鼠急性肺损伤模型,C组给予等容量生理盐水,MK组和MS组于注射内毒素前30 min经尾静脉分别注射K252a 75μg/kg、SB203580 10 mg/kg.ALI组、MK组和MS组于注射内毒素后1、3、6、12 h(1-4)时各组随机取6只大鼠,C组于给予生理盐水后即刻处死取肺,采用ELISA法测定支气管肺泡灌洗液中TNF-α浓度,称重后计算肺湿干重比,采用Western b1ot法测定p-MLK3、p-MKK3/6及p-p38MAPK的表达,观察肺组织病理学结果.结果 与C组比较,ALI组、MK组和MS组各时点支气管肺泡灌洗液中TNF-α浓度、肺湿干重比、p-MLK3、p-MKK3/6及p-p38MAPK的表达水平升高(P＜0.01);与ALI组比较,MK组上述指标降低,MS组支气管肺泡灌洗液中TNF-α浓度、肺湿干重比、p-p38MAPK表达水平降低(P＜0.05).病理学结果显示:MK组和MS组肺组织损伤较ALI组减轻.结论 MLK3-MKK3/6-p38MAPK信号转导通路在大鼠内毒素性急性肺损伤中起重要作用.     

P38MAPK信号通路在糖尿病肾脏病研究中的进展

糖尿病肾脏病是糖尿病最严重的慢性并发症之一,是导致慢性肾衰的主要病因,P38MAPK是MAPK信号通路中重要的信号转导分子,是细胞信号传递的交汇点和共同通路.p38MAPK信号转导通路可通过增加活性氧类的产生增加炎性介质的释放,调节肾素-血管紧张素系统,影响肾小球系膜外基质的形成与降解,从而加速糖尿病肾脏病进程,以p3...     

Diacylglycerol production and protein kinase C activity are increased in a mouse model of diabetic embryopathy

Activation of the diacylglycerol-protein kinase C (DAG-PKC) cascade by excess glucose has been implicated in vascular complications of diabetes. Its involvement in diabetic embryopathy has not been established. We examined DAG production and PKC activities in embryos and decidua of streptozotocin (STZ)-diabetic or transiently hyperglycemic mice during neural tube formation. STZ diabetes significantly increased DAG and total PKC activity in decidua (1.5- and 1.4-fold, respectively) and embryos (1.7- and 1.3-fold, respectively) on day 9.5. Membrane-associated PKC alpha, betaII, delta, and zeta were increased in decidua by 1.25- to 2.8-fold. Maternal hyperglycemia induced by glucose injection on day 7.5, the day before the onset of neural tube formation, also increased DAG, PKC activity, and PKC isoforms (1.1-, 1.6-, and 1.5-fold, respectively) in the embryo on day 9.5. Notably, membrane-associated PKC activity was increased 24-fold in embryos of diabetic mice with structural defects. These data indicate that hyperglycemia just before organogenesis activates the DAG-PKC cascade and is correlated with congenital defects.     

p38丝裂原活化蛋白激酶与病理性疼痛

病理性疼痛病理机制复杂,临床常用的药物治疗效果差。p38丝裂原活化蛋白激酶可通过多种方式影响疼痛的形成与维持。动物试验及部分临床研究初步表明,p38MAPK特异性的抑制剂用于治疗病理性疼痛可能具有良好的应用前景。     

p38MAPK信号通路在鞘内注射利多卡因诱发糖尿病神经病变大鼠脊髓神经元凋亡中的作用

目的探讨p38丝裂原活化蛋白激酶(p38MAPK)信号通路在鞘内注射利多卡因诱发糖尿病(DM)神经病变(DNP)大鼠脊髓神经元凋亡中的作用。方法健康成年雄性SD大鼠50只,随机取10只为对照组(C组),余40只大鼠高糖高脂饮食+小剂量链脲佐菌素(STZ)腹腔注射建立2型DM,之后继续喂养28d,得35只DNP大鼠。将C组大鼠和DNP大鼠均进行鞘内置管。将置管成功的25只DNP大鼠采用随机数字法分为三组:NS组8只,鞘内注射重比重利多卡因10μl 3d+0.9%NaCl 10μl 5d;DM组8只,鞘内注射重比重利多卡因10μl 3d+2%二甲亚砜(DMSO)10μl 5d;SB组9只,鞘内注射重比重利多卡因10μl 3d+SB203580(SB203580需溶解在DMSO溶剂中)10μg/10μl 5d。于腹腔注射STZ前(C组鞘内置管前28d,T1)、STZ后28d(C组鞘内置管前,T2)、STZ后34d(T3)、STZ后39d(T4)时测定大鼠后爪机械缩足反射阈值(MWT);测完MWT立即处死大鼠,取L4~5的脊髓组织,光镜下观察其病理学结果,采用TUNEL法检测脊髓神经元凋亡的情况,采用ELISA法检测p-p38MAPK水平。结果 T2、T3时NS、DM和SB组MWT均明显低于C组(P0.05)。T4时NS和DM组MWT明显低于C和SB组(P0.05)。NS和DM组脊髓神经元凋亡指数,脊髓p-p38MAPK水平明显高于C和SB组(P0.05)。HE染色光镜下见NS组及DM组大鼠脊髓组织结构模糊,出现轻度的水肿,同时神经元的细胞核间隙轻微增宽;C组脊髓组织无明显病理改变;SB组大鼠脊髓组织病理损伤较轻,几乎正常。结论鞘内注射重比重利多卡因诱发DNP大鼠脊髓神经元的凋亡可能与进一步激活p38MAPK信号通路有关,且应用p38MAPK抑制剂SB203580对其有保护作用。