中医药与p38丝裂原活化蛋白激酶信号转导通路研究

丝裂原活化蛋白激酶(MAPK)信号通路广泛存在于哺乳动物细胞中,与机体多种生理病理过程(如糖尿病,心血管疾病,脑发育等)密切相关,是20世纪90年代以来生命科学研究的热点。MAPK分为4个家族,即细胞外调节激酶、c-jun氨基末端激酶、p38和大丝裂原活化蛋白激酶1/细胞外调节激酶5。综述近几年来从p38 MAPK信号通路角度进行的中医药研究,提示从细胞信号转导角度进行中医药研究有着十分广阔的发展前景。     

p38丝裂原活化蛋白激酶通路及其研究进展

丝裂原活化蛋白激酶(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。它通过转录因子磷酸化而改变基因的表达水平,参与多种胞内信息传递过程,能对广泛的细胞外刺激发生反应,介导细胞生长、发育、分化及死亡全过程。近年研究发现,p38MAPK在许多疾病的发病过程中具有重要作用。     

p38丝裂原活化蛋白激酶与细胞迁移

丝裂原活化蛋白激酶(mitogen activated protein kinases,MAPKs)是一类存在于大多数真核细胞内,转导胞外信号引起细胞反应的丝/苏氨酸蛋白激酶,是细胞内一重要信号系统.其中,p38MAPK信号通路是MAPKs家族的重要组成部分,它经外界刺激应激而激活,故又称为MAPK应急信号通路,其在全身炎性反应、细胞分化及凋亡等方面具有十分重要的作用,近年研究发现p38MAPK信号通路也参与细胞迁移的调控.现就p38MAPK及其在细胞迁移中的作用的研究进展进行综述.     

p38丝裂原活化蛋白激酶信号级联在炎症反应中的作用

在急、慢性疾病中，细胞释放的炎症介质可以活化多种信号转导级联反应，丝裂原活化蛋白激酶(MAPK)信号转导通道在招募白细胞于炎症部位聚集起着重要的作用。同时，p38MAPK异构体的活化可以激活致炎细胞因子，而后刺激白细胞活化。然而，p38MAPK引起的白细胞招募这一系列的功能过程包括：粘附、游走和效应器的功能如氧化爆发以及p38MAPK介导的复杂细胞因子网络在炎症中的作用仍有待研究。针对减少炎症介质产生和以p38MAPK为治疗靶点的研究正在进行中，不远的将来可能会成为治疗炎症疾病的新策略。     

p38丝裂原活化蛋白激酶在雄性小鼠生殖细胞株中的表达

目的 检测p38 丝裂原活化蛋白激酶(p38 mitogen-activated protein kinase,p38 MAPK)在雄性小鼠生殖细胞株TM4、GC-1spg、GC-2spd(ts)中的表达,为深入研究p38 MAPK基因在雄性小鼠精子发生中的作用机制奠定理论基础.方法 应用逆转录-多聚合酶链反应(RT-PCR)检测雄性小鼠生殖细胞株TM4、GC-1spg、GC-2spd(ts)中p38 MAPK基因的表达.结果 p38 MAPK基因在TM4、GC-1spg、GC-2spd(ts)细胞株中均为阳性表达.结论 p38 MAPK基因的表达在雄性小鼠精子发生中可能发挥重要作用.     

p38丝裂原活化蛋白激酶在糖尿病肾脏疾病中的研究进展

p38丝裂原活化蛋白激酶(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或共同通路,可被磷酸化为p-p38MAPK而激活,参与细胞增殖、生长、分化及凋亡等多种细胞行为的调控。最近的研究显示,p38MAPK信号转导通路可能通过调节转化生长因子β1的转录与合成,调节炎性因子的表达,活化环腺苷酸反应原件结合蛋白等转录因子,加重肾脏的炎症和纤维化进程,从而加速糖尿病肾病进程。     

p38丝裂原活化蛋白激酶信号通路在丙泊酚器官保护中的作用

丝裂原活化蛋白激酶(MAPK)在对细胞外刺激因素进行应答时发挥着重要作用,介导多种细胞行为的变化。其中,p38信号通路是MAPK家族中关键通路之一,其介导细胞生长、发育、分化及死亡的全过程。丙泊酚除具有麻醉作用外,还可作用于一些MAPK,包括细胞外信号调节激酶、c-Jun氨基末端激酶和p38MAPK,从而发挥抗肿瘤、抗炎、抗氧化和抗凋亡等积极作用。该文通过总结p38MAPK信号通路在丙泊酚对器官保护中所发挥的作用,为丙泊酚的临床器官保护作用机制提供一种新的研究思路。     

丝裂原活化蛋白激酶p38在急性胰腺炎患者外周血单个核细胞中的变化和意义

目的探讨急性胰腺炎患者外周血单个核细胞(PBMC)内丝裂原活化蛋白激酶p38(p38 MAPK)信号水平的变化及其与疾病严重程度的关系。方法收集轻型胰腺炎(MAP组)29例、重症胰腺炎(SAP组)23例和对照组21例,实时荧光聚合酶链反应检测患者PBMC p38 MAPK基因表达水平,Western blot检测PBMC p38 MAPK和磷酸化p38 MAPK(P-p38 MAPK)蛋白的水平。结果 SAP组p38 MAPK mRNA表达水平和总蛋白水平高于对照组和MAP组(P<0.05);MAP组p38 MAPK mRNA表达水平和总蛋白水平与对照组比较差异无统计学意义(P>0.05)。SAP组P-p38 MAPK总蛋白水平高于对照组和MAP组(P<0.01,P<0.05),MAP组P-p38 MAPK总蛋白水平高于对照组(P<0.05)。结论急性胰腺炎患者PBMC内p38 MAPK mRNA表达水平和总蛋白水平变化较小;p38 MAPK磷酸化水平显著升高,而且与病情程度密切相关。     

p38丝裂原活化蛋白激酶信号通路对胰岛素作用的研究进展

丝裂原活化蛋白激酶(MAPK)在哺乳动物细胞多种信号转导通路中起重要作用,p38MAPK作为其成员之一,可以调节细胞的多种生物学反应,在2型糖尿病胰岛素分泌和胰岛素抵抗发病机制中有重要的作用。p38δ-蛋白激酶D通路综合调节胰岛素分泌能力和胰腺β细胞的存活,p38 MAPK信号通路通过炎性因子、游离脂肪酸、氧化应激等对脂肪、骨骼肌作用,参与胰岛素抵抗。     

p38丝裂原活化蛋白激酶与骨代谢

丝裂原活化蛋白激酶(mitogen-activated protein kinases,MAPK)是机体广泛表达的丝氨酸/酪氨酸激酶,在哺乳动物细胞多种信号转导通路中起重要作用。p38MAPK信号通路是MAPK通路的一个重要分支,在细胞增殖、分化、凋亡和细胞周期调控等多种生理和病理过程中发挥重要作用。近年来,有关p38MAPK信号通路在与骨代谢相关的破骨细胞、成骨细胞、软骨细胞生长、代谢及功能方面的研究倍受关注。本文就 p38MAPK与骨代谢相关研究进展进行综述,旨在探讨p38MAPK在骨代谢相关疾病中的作用机制。     

基于细胞穿透肽技术对p38丝裂原活化蛋白激酶蛋白功能的研究

目的 构建基于TAT技术的p38 MAPK蛋白转运系统并对导入细胞的融合蛋白的功能进行鉴定。方法 采用亚克隆方法构建重组质粒pHis-TAT-p38和pHis-TAT-p38（AF）无活性突变体,并诱导原核表达纯化融合蛋白;将这两种蛋白加入培养的ECV304细胞,在高渗刺激下,通过检测p38底物ATF2的磷酸化水平,以观察由TAT转导进入细胞His-TAT-p38及其无活性突变体对内源性p38活性的影响。结果 酶切和测序结果表明,载体构建正确;SDS-PAGE凝胶电泳可见原核表达纯化得到高纯度目的蛋白：Western blot表明,融合蛋白His-TAT-p38及其突变体能够以一种时间和浓度依赖性方式高效转导进入细胞：导入His-TAT-p38蛋白后,结果发现导入的野生型p38可增强内源性p38的功能,而无活性突变体则竞争性抑制了内源性p38MAPK蛋白的功能,从而阻止或抑制其对内源性底物ATF2的磷酸化．使得信号不能传递下去,进而抑制p38MAPK通路的活动。结论 成功建立了基于TAT的蛋白转运系统．并证实TAT能够以浓度和时间依赖方式高效转运蛋白进入真核细胞;进入细胞的His-TAT-p38和His-TAT-p38无活性突变体蛋白均具有较高的生物学活性,在高渗刺激作用下前者可以增加p38磷酸化水平,而后者则显著抑制p38信号通路的活性。     

p38MAPK信号通路与心脏重构关系的研究进展

心脏重构是一种常见的病理生理状态,包括心肌肥厚及间质纤维化,是各种心血管疾病发展为慢性心功能不全、心力衰竭的重要环节,由多种体内外刺激因素通过启动细胞内共同信号转导通路而促发/激活。有研究表明,p38丝裂原蛋白活化激酶(MAPK)激活参与心脏重构并在其中起着重要作用,用特异性抑制剂阻断该信号通路可明显减轻心脏重构。该文就p38MAPK对心脏重构重要环节影响的研究进展予以综述。     

p38蛋白激酶(p38 MAPK)在癫疒间大鼠脑内的表达

 目的  观察p38蛋白激酶(p38 mitogen-activated protein kinase,p38 MAPK)在癫疒间大鼠脑内的表达情况。方法  健康雄性SD大鼠随机分成正常对照组(n=8)和癫疒间组(n=8)。采用戊四氮腹腔注射建立癫疒间模型,大鼠点燃后的惊厥行为按照Racine的标准进行观察评分,采用Western blot和免疫荧光法比较两组大鼠脑内p38 MAPK的表达情况。结果  癫疒间组大鼠脑内p38 MAPK在皮层和海马的表达均显著高于正常对照组(P<0.01)。结论  p38 MAPK在癫疒间大鼠脑内表达上调。     

高氧对幼鼠肺组织细胞凋亡及p38 MAPK丝裂原活化蛋白激酶的影响

目的 探讨高氧对幼鼠肺组织细胞凋亡及p38 MAPK丝裂原活化蛋白激酶表达的影响.方法 幼年Wistar大鼠90%氧气暴露建立高氧肺损伤模型,行肺组织病理学检查,应用TUNEL法检测肺组织的细胞凋亡,Western blot法检测p38 MAPK表达.结果 高氧暴露3 d可见肺组织水肿、出血、炎症细胞浸润等急性肺损伤改变.高氧3 d组的肺凋亡指数较空气对照组明显增加,凋亡发生的主要部位为肺泡、支气管上皮及血管内皮细胞.Westem blot结果显示高氧暴露1 d,肺组织的p38MAPK活性迅速升高,于第2天达到高峰,第3天活性有所下降,但仍高于空气对照组.结论 高氧暴露可以诱导肺组织发生细胞凋亡;高氧可以激活p38MAPK通路,参与高氧性肺损伤的调控;活化的p38MAPK可能参与了高氧诱导的肺组织细胞凋亡的调控.     

高血压血管重构的中西医研究进展

血管重构是近年来高血压病的研究热点之一,在此通过阐述异常血流动力学、血管活性物质、细胞凋亡、细胞外基质等因子参与高血压血管重构的机制以及中西医药物在血管重构中的治疗作用,综述目前高血压血管重构的中西医研究进展,为中西医结合治疗高血压血管重构提供思路。     

高温高湿环境颅脑火器伤后兔脑p38MAPK变化

目的 研究p38MAPK在高温高湿环境下颅脑火器伤后活化及其原因。方法 采用高温高湿颅脑枪伤模型,新西兰大白兔30只,随机分成常温[(22.0±0.5)℃、RH50%]对照组,高温高湿[(39.0±0.5)℃、RH80%~85%]枪伤后受热10和30min,1h、1.5h、2h组,每组5只。采用Westernblot检测脑匀浆p38MAPK活性,应用化学发光和X线片显示,凝胶图像分析仪半定量分析。结果 颅脑枪伤受高温高湿环境作用脑皮质和下丘脑的p38MAPK的活性受湿热后迅速增高,于受热1h达高峰,随后下降。下丘脑的p38MAPK活性比脑皮质高。结论 高温高湿环境颅脑火器伤后p38MAPK活性早期明显升高,参与了脑的继发性损害过程。     

高温高湿环境颅脑火器伤后兔脑p38MAPK变化

OBJECTIVE: To study the activation of p38 mitogen-activated protein kinase (MAPK) in the brain of rabbits after craniocerebral gunshot injury in a hot and humid environment (HHE) and explore its possible mechanism. METHODS: Craniocerebral gunshot injury model was established in 30 New Zealand white rabbits, which were subsequently exposed to environment of normal temperature (at 22.0% +/- 0.5 degrees C; with relative humidity of 50%) or HHE at 39.0 +/- 0.5 degrees C; with relative humidity of 80%-85% for 10 min, 30 min, 1 h, 1.5 h, and 2 h groups, respectively, with 5 rabbits in each group. p38 MAPK activity in the brain tissues of the rabbits following the injury and environmental exposure were detected by Western blotting and analyzed semi-quantitatively by Bio-Profil gel image analysis system. RESULTS: p38 MAPK activity in the cortex and hypothalamus was significantly elevated following gunshot injury and HHE exposure, reaching the peak level at 1 h of HHE exposure and then decreased. p38 MAPK activity was significantly higher in the hypothalamus than in the cortex. CONCLUSION: p38 MAPK activity increases in the early stage following craniocerebral gunshot injury and HHE exposure in rabbits, the mechanism of which might involve the secondary brain insult.     

The Relationship between p38MAPK and Apoptosis during Paclitaxel Resistance of Ovarian Cancer Cells

To investigate the relationship between p38 mitogen-activated protein kinase (p38MAPK) and cell apoptosis during the paclitaxel resistance of ovarian carcinoma cell lines, flow cytometry (FCM) and PI staining were employed to determine the effect of p38MAPK inhibitor SB203580 on the apoptosis of A2780/Taxol cells, a drug-resistant human ovarian carcinoma cell line. p38MAPK protein expression in SB203580-treated cells was immunochemically measured. The 50% inhibition concentration (IC50) of paclitaxel on A2780/Taxol cells was determined by MTT assay. MDR-1 mRNA, and expression of p38MAPK and phospho-p53 protein were detected by RT-PCR and West- ern blotting, respectively. The apoptosis rate of A2780/Taxol cells was (19.7±1.04)% 24 h after SB203580 treatment. A significant difference in apoptosis rate was found among experiment group, control group and untreated group (P<0.05). The relative reversal rate of A2780/Taxol cells to pacli- taxel was (57.18±2.01)%. As compared with the control group and the untreated group, p38MAPK protein and MDR-1 mRNA in SB203580-treated cells was substantially decreased. The expression of p53 protein was significantly increased. It is concluded that p38MAPK pathway is related to pacli- taxel resistance of ovarian carcinoma, and blockade of this pathway can promote the apoptosis of the drug-resistant cells and reverse the drug-resistance. Moreover, p38MAPK-mediated apoptosis in pa- clitaxel-resistant ovarian carcinoma cells depends on the activation of p53.     

The Relationship between p38MAPK and Apoptosis during Paclitaxei Resistance of Ovarian Cancer Cells

To investigate the relationship between p38 mitogen-activated protein kinase (p38MAPK) and cell apoptosis during the paclitaxel resistance of ovarian carcinoma cell lines, flow cytometry (FCM) and PI staining were employed to determine the effect of p38MAPK inhibitor SB203580 on the apoptosis of A2780/Taxol cells, a drug-resistant human ovarian carcinoma cell line. p38MAPK protein expression in SB203580-treated cells was immunochemically measured. The 50% inhibition concentration (IC(50)) of paclitaxel on A2780/Taxol cells was determined by MTT assay. MDR-1 mRNA, and expression of p38MAPK and phospho-p53 protein were detected by RT-PCR and Western blotting, respectively. The apoptosis rate of A2780/Taxol cells was (19.7+/-1.04)% 24 h after SB203580 treatment. A significant difference in apoptosis rate was found among experiment group, control group and untreated group (P<0.05). The relative reversal rate of A2780/Taxol cells to paclitaxel was (57.18+/-2.01)%. As compared with the control group and the untreated group, p38MAPK protein and MDR-1 mRNA in SB203580-treated cells was substantially decreased. The expression of p53 protein was significantly increased. It is concluded that p38MAPK pathway is related to paclitaxel resistance of ovarian carcinoma, and blockade of this pathway can promote the apoptosis of the drug-resistant cells and reverse the drug-resistance. Moreover, p38MAPK-mediated apoptosis in paclitaxel-resistant ovarian carcinoma cells depends on the activation of p53.     

The relationship between p38MAPK and apoptosis during paclitaxel resistance of ovarian cancer cells

Summary To investigate the relationship between p38 mitogen-activated protein kinase (p38MAPK) and cell apoptosis during the paclitaxel resistance of ovarian carcinoma cell lines, flow cytometry (FCM) and PI staining were employed to determine the effect of p38MAPK inhibitor SB203580 on the apoptosis of A2780/Taxol cells, a drug-resistant human ovarian carcinoma cell line. p38MAPK protein expression in SB203580-treated cells was immunochemically measured. The 50% inhibition concentration (IC₅₀) of paclitaxel on A2780/Taxol cells was determined by MTT assay. MDR-1 mRNA, and expression of p38MAPK and phospho-p53 protein were detected by RT-PCR and Western blotting, respectively. The apoptosis rate of A2780/Taxol cells was (19.7±1.04)% 24 h after SB203580 treatment. A significant difference in apoptosis rate was found among experiment group, control group and untreated group (P<0.05). The relative reversal rate of A2780/Taxol cells to paclitaxel was (57.18±2.01)%. As compared with the control group and the untreated group, p38MAPK protein and MDR-1 mRNA in SB203580-treated cells was substantially decreased. The expression of p53 protein was significantly increased. It is concluded that p38MAPK pathway is related to paclitaxel resistance of ovarian carcinoma, and blockade of this pathway can promote the apoptosis of the drug-resistant cells and reverse the drug-resistance. Moreover, p38MAPK-mediated apoptosis in paclitaxel-resistant ovarian carcinoma cells depends on the activation of p53. This project was supported by a grant from R&D program of Heilongjiang Province (No. GB05C402-11).