p38MAPK信号转导通路与肿瘤关系的最新研究进展

丝裂原活化蛋白激酶(MAPK)级联是细胞内广泛存在的一类丝/苏氨酸蛋白激酶超家族,其亚族主要包括细胞外信号调节激酶(ERK)、c-Jun氨基末端激酶/应激激活蛋白激酶(JNK/SAPK)和p38MAPK.他们主要是将细胞外信号转入细胞核内并引起相应变化的重要信号通路,调节细胞生长、分化、对环境的应激适应、炎症反应等多种...     

The priming effect of C5a on monocytes is predominantly mediated by the p38 MAPK pathway

The dysregulation of the inflammatory response after trauma leads to significant morbidity and mortality. Monocytes and macrophages play a central role in the orchestration of the inflammatory response after injury. Serum interleukin-6 (IL-6) concentration correlates with poor outcomes after injury. Tumor necrosis factor-alpha (TNF-alpha) is a proinflammatory cytokine that plays a crucial role in the pathogenesis of multiple organ dysfunction syndrome. Furthermore, in the presence of C5a, monocytes and macrophages have potentiated responses, but the mechanisms underlying this response remain largely unknown. Peripheral blood mononuclear cells (PBMCs) were isolated from healthy volunteers and pretreated with C5a (100 ng/mL) for 1 h before adding lipopolysaccharide (LPS) (10 ng/mL) for up to 20 h. Inhibitors for the mitogen-activated protein kinases (MAPKs) were added 1 h before adding C5a. C5a primes monocytes for LPS-induced IL-6 and TNF-alpha production. Treatment of PBMCs with C5a leads to a rapid activation of the 3 MAPK pathways. SP600125 (inhibitor of c-Jun NH2-terminal kinase MAPK) and PD98059 (inhibitor of extracellular signal-regulated kinase MAPK) did not affect the C5a priming of the LPS-induced IL-6 and TNF-alpha production, whereas SB203580, a specific inhibitor of p38 MAPK, did suppress the C5a priming effect. These results demonstrate that C5a primes adherent PBMCs and modulates LPS-induced IL-6 and TNF-alpha production. Results from extracellular signal-regulated kinase and c-Jun NH2-terminal kinase MAPK blockade suggest that these signaling pathways have minimal or no role in reprogramming LPS-mediated IL-6 and TNF-alpha production. On the contrary, in PBMCs, C5a activates the p38 cascade, and this pathway plays a major role in the C5a enhancement of LPS-induced IL-6 and TNF-alpha production.     

Interleukin-1beta-induced transdifferentiation of renal proximal tubular cells is mediated by activation of JNK and p38 MAPK

Interleukin (IL)-1beta induces renal tubular epithelial cells to transdifferentiate to myofibroblasts, which express alpha-smooth muscle actin (alpha-SMA). To understand the signal transduction mechanisms involved in transdifferentiation, we examined the roles of mitogen-activated protein kinases (MAPKs) in IL-1beta-stimulated alpha-SMA expression and cell migration in the HK-2 human renal proximal tubular cell line. IL-1beta induced the transdifferentiation of renal proximal tubular cells, which was characterized by upregulated expression of alpha-SMA and increased cell migration. In addition, IL-1beta increased the activity of the three members of the MAPK family, ERK, JNK and p38 MAPK, in these cells. Both SP600125, a specific inhibitor of JNK, and SB203580, a specific inhibitor of p38 MAPK, suppressed the IL-1beta-induced expression of alpha-SMA and cell migration, but these effects were not observed with PD98059, a specific inhibitor of ERK. These results suggest that IL-1beta-induced HK-2 cell transdifferentiation is mediated, at least in part, through the activation of the JNK and p38 MAPK signaling pathways.     

应激后大鼠下丘脑磷酸化p38MAPK的变化及电针足三里穴的调节作用

目的 观察应激对大鼠下丘脑磷酸化p38MAPK的影响及电针足三里穴的调节作用。方法将28只Wistar大鼠随机分为对照组、束缚应激组和束缚应激 电针组，后2组再根据束缚解除后及电针足三里穴位治疗后不同时间点分别分为3个亚组。电针治疗采用频率为5～12Hz的疏密波，治疗总时间30min。于相应时间点处死各组大鼠，采用Western印迹杂交法观察其下丘脑内磷酸化p38MAPK的变化。结果束缚应激组大鼠下丘脑内p38MAPK磷酸化水平明显高于对照组，且在束缚应激解除后3h仍保持在较高水平；应激大鼠经电针足三里穴处理后，下丘脑内磷酸化p38MAPK与束缚应激组相比明显回落，这种调节作用可持续到电针治疗结束后3h。结论束缚应激能引起下丘脑应激信号转导系统中p38MAPK的磷酸化，这可能是应激活动过程中的重要事件之一。电针足三里穴对下丘脑内磷酸化p38MAPK的调节作用可能与下丘脑-垂体-肾上腺轴的活动有关。     

抑制p38 MAPK通路对大鼠癫痫发作引起海马神经元损伤的保护作用

目的探讨抑制p38 MAPK通路对减轻红藻氨酸诱导的颞叶癫痫发作引起大鼠海马神经元所造成损害的作用和机制.方法经侧脑室给予不同剂量(0,0.01,0,1,1 μg)SB203580(p38 MAPK特异性抑制剂)预处理,30 min后再予红藻氨酸制作大鼠颞叶癫痫持续状态模型.7 d后采用Nissl染色方法观察海马各区的锥体细胞和颗粒细胞的情况.结果红藻氨酸注射7 d后CA3区可见大量锥体细胞缺失,尚存细胞变性,伴局限性颗粒细胞增多,呈放射状分布.CA1区受累较轻,但也存在一定程度的细胞脱失.预先给予SB203580(0.1μg以上剂量)处理的动物以上变化明显减轻.假手术大鼠海马各区有大量正常致密锥体细胞,未见细胞脱失.结论大鼠侧脑室内注射红藻氨酸引起全身痉挛性癫痫持续状态对海马神经元造成损害,而抑制p38 MAPK通路,对海马神经元起一定保护作用.     

Ligustrazine alleviates acute pancreatitis by accelerating acinar cell apoptosis at early phase via the suppression of p38 and Erk MAPK pathways

ObjectiveThis study aimed to investigate the role of ligustrazine on apoptosis and inflammatory reaction in acute pancreatitis.MethodsRats and acinar cells were treated with caerulein to induce acute pancreatitis models. Cell models were treated with saline, p38 inhibitor, Erk inhibitor and ligustrazine. Then, the levels of TNF-α, IL-1β and IL-6 were determined by ELISA assay, the protein levels of p38, Erk1/2, p53 and cleaved caspase3 were determined by western blotting, and apoptosis were measured by flow cytometry. Rat models were treated with saline and ligustrazine. Plasma amylase and pancreatic myeloperoxidase activity and the levels of TNF-α, IL-1β and IL-6 in rats were determined. The protein levels of p38, Erk1/2, p53 and cleaved caspase3 in pancreas tissues were determined by western blotting, and pancreas tissues were also performed TUNEL staining to observe apoptosis status.ResultsLigustrazine downregulated the levels of TNF-α, IL-1β, IL-6. The protein levels of p38 and Erk were reduced by p38 inhibitor, Erk inhibitor and ligustrazine, while the levels of p53 and cleaved caspase 3 were upregulated. Apoptosis of AP acinar cells and cells in AP rat models was promoted after treated with ligustrazine. Plasma amylase and pancreatic myeloperoxidase activity in AP rat models were reduced by ligustrazine.ConclusionLigustrazine alleviates acute pancreatitis by accelerating acinar cell apoptosis at early phase via the suppression on p38 and Erk MAPK pathways. It is capable of attenuating the severity of acute pancreatitis and may have a therapeutic effect on patients with acute pancreatitis.     

p38 MAPK蛋白在米诺环素抑制小鼠EAE发病中的表达变化及意义

【目的】探讨米诺环素抑制小鼠实验性变态反应性脑脊髓炎（EAE）发病过程中，p38 MAPK蛋白的表达变化及意义。【方法】将小鼠随机分为对照组、EAE模型纽和米诺环素治疗纽。在动物发病高峰期取小鼠脑组织和脊髓，行HE组织染色观察病理改变，以及Western Blot方法检测p-p38MAPK蛋白的表达。【结果】①EAE组脑组织和脊髓有多部位的血管炎，血管周围大量炎性细胞浸润，小血管明显扩张，改变以白质为著。治疗组的血管炎范围、数量及程度均明显减轻。②p-p38 MAPK蛋白在对照组仅有少量表达；在EAE组表达明显升高，达3倍左右；治疗组的表达显著降低，但仍较对照组为高。【结论】p38 MAPK途径可能参与了米诺环素对小鼠EAE发病过程的抑制作用。     

p38MAPK信号通路与应力介导成肌细胞的凋亡

背景：在体内条件下,细胞力学的功能研究因其所处生理环境的复杂性、实验条件的不易控制而很难得到满意结果。目的：在成功构建成肌细胞体外培养-力学刺激模型的基础上,研究p38MAPK信号通路在成肌细胞凋亡中的作用及其机制。方法：将体外培养的C2C12细胞分为对照组和SB203580组,SB203580组中加入20mmol/L的p38MAPK抑制剂SB203580。应用细胞应力加载装置FlecellStrainUnit-5000T给细胞提供15%的力值,分别施加0,6,12,24h的周期性张应力。每分钟10个循环,每循环包括3s牵张,3s松弛。Hoechst33258染色观察细胞的形态学变化;流式细胞仪检测细胞凋亡情况;RT-PCR法检测促凋亡基因baxmRNA的表达;Westernblot检测信号通路中p38MAPK和p-p38MAPK蛋白的表达。结果与结论：随着加力时间的延长,细胞逐渐出现核固缩及凋亡小体,凋亡率增加（P〈0.05）,baxmRNA表达增多（P〈0.05）;细胞p38MAPK和p-p38MAPK蛋白均在加力6h达到最低,此后逐渐升高。p38MAPK抑制剂SB203580可抑制加力引起的细胞凋亡,减少baxmRNA及p38MAPK和p-p38MAPK蛋白的表达（P〈0.05）。说明p38MAPK信号通路在应力介导的成肌细胞凋亡中起到重要的作用。     

p38MAPK信号通路与应力介导成肌细胞的凋亡(英文)

背景:在体内条件下,细胞力学的功能研究因其所处生理环境的复杂性、实验条件的不易控制而很难得到满意结果。目的:在成功构建成肌细胞体外培养-力学刺激模型的基础上,研究p38MAPK信号通路在成肌细胞凋亡中的作用及其机制。方法:将体外培养的C2C12细胞分为对照组和SB203580组,SB203580组中加入20mmol/L的p38MAPK抑制剂SB203580。应用细胞应力加载装置FlecellStrainUnit-5000T给细胞提供15%的力值,分别施加0,6,12,24h的周期性张应力。每分钟10个循环,每循环包括3s牵张,3s松弛。Hoechst33258染色观察细胞的形态学变化;流式细胞仪检测细胞凋亡情况;RT-PCR法检测促凋亡基因baxmRNA的表达;Westernblot检测信号通路中p38MAPK和p-p38MAPK蛋白的表达。结果与结论:随着加力时间的延长,细胞逐渐出现核固缩及凋亡小体,凋亡率增加(P<0.05),baxmRNA表达增多(P<0.05);细胞p38MAPK和p-p38MAPK蛋白均在加力6h达到最低,此后逐渐升高。p38MAPK抑制剂SB203580可抑制加力引起的细胞凋亡,减少baxmRNA及p38MAPK和p-p38MAPK蛋白的表达(P<0.05)。说明p38MAPK信号通路在应力介导的成肌细胞凋亡中起到重要的作用。     

拓朴替康诱导K562细胞凋亡时p38MAPK的变化

目的:探讨拓朴替康诱导K562细胞凋亡的相关信号转导途径。方法:0.1μmol/L的拓朴替康处理K562细胞24h时,通过流式细胞仪(FCM)和片段DNA含量检测研究拓朴替康对靶细胞的促凋亡活性;采用免疫印迹法和γ-32P掺入法测定拓朴替康对p38MAPK含量及其活性的影响。结果:FCM检测发现在K562细胞周期图中出现一凋亡峰,凋亡细胞比率为16.9%,凋亡细胞特有的片段DNA含量为(45.9±1.0)%;p38MAPK含量及其活性明显升高,分别为对照组的4.9倍和4.1倍。结论:拓朴替康诱导K562细胞凋亡可能通过激活p38MAPK来实现。     

l-Menthol alleviates cigarette smoke extract induced lung injury in rats by inhibiting oxidative stress and inflammation via nuclear factor kappa B,p38 MAPK and Nrf2 signalling pathways

l-Menthol is the main ingredient of peppermint which affects various pharmacological effects such as anti-inflammation and anti-oxidative activity. In this study, we aimed to evaluate the potential effects of l-menthol on cigarette smoke extract (CSE) induced lung injury in rats. Morphology assessment results revealed that administration with l-menthol (5, 10 or 20 mg kg⁻¹ d⁻¹) significantly alleviated CSE-induced lung injury. Besides, l-menthol significantly reduced the inflammatory response by suppressing the production of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and interleukin-6 (IL-6) via downregulating nuclear factor kappa B (NF-κB) and p38 MAPK pathways. Meanwhile, l-menthol decreased the levels of oxidative stress markers including malondialdehyde (MDA) and myeloperoxidase (MPO) whereas it increased the amount of glutathione (GSH), glutathione peroxidase (GSH-Px), superoxide dismutase (SOD) and total antioxidant capacity (T-AOC) through activation of the Nrf2 pathway. Furthermore, the expression of MMP-9 and TIMP-1 in lungs was reduced after treatment with l-menthol, and this indicated that l-menthol might have a potential effect on airway remodeling. Moreover, immunohistochemistry analyses indicated that l-menthol could suppress the infiltration of CD4⁺ and CD8⁺ T cells in lung tissues and this was probably due to the immune regulation activity of l-menthol. Taken together, our findings support that l-menthol might be a potential candidate for the treatment of CSE-induced lung injury in rats.

l-Menthol is the main ingredient of peppermint which affects various pharmacological effects such as anti-inflammation and anti-oxidative activity.     

法舒地尔对脂多糖诱导急性肺损伤大鼠肺组织p38丝裂原活化蛋白激酶激活的影响

目的观察法舒地尔对脂多糖(LPS)诱导的急性肺损伤(ALI)大鼠肺组织p38MAPK磷酸化激活的影响。方法 45只雄性SD大鼠随机分为三组:对照组(NS组,n=15)、脂多糖组(LPS组,n=15)及法舒地尔干预组(FAS组,n=15),LPS组和FAS组采用股静脉注射LPS(5 mg/kg)建立ALI模型,FAS组在LPS注射前30 min腹腔注射法舒地尔(10 mg/kg)。造模后3 h、6 h、12 h观察肺组织病理形态学改变,免疫组织化学及Western Blot法检测肺组织p-p38MAPK的表达。结果造模后LPS组各时间点肺组织p-p38MAPK的表达较NS组明显增加(P<0.05),与LPS组相比,FAS组各时间点p-p38MAPK的表达明显减少(P<0.05)。光镜显示LPS组病理形态学改变明显,而FAS组则明显减轻。结论法舒地尔可明显抑制脂多糖诱导的急性肺损伤大鼠肺组织p38MAPK的磷酸化激活,减轻脂多糖诱导的大鼠肺损伤程度。     

Paradoxical effect of caspofungin against Candida bloodstream isolates is mediated by multiple pathways but eliminated in human serum

Paradoxical growth of Candida in vitro at echinocandin concentrations exceeding the MIC is well described, but the clinical relevance is unknown. We assessed echinocandin paradoxical effects against Candida bloodstream isolates (BSI) in the presence or absence of human serum and investigated regulatory mechanisms. As determined by broth microdilution, a paradoxical effect was evident for 60% (18/30), 23% (7/30), and 13% (4/30) of Candida albicans BSI exposed to caspofungin, anidulafungin, and micafungin, respectively, at achievable human serum concentrations (≤8 μg/ml). A paradoxical effect was not evident among 34 C. glabrata BSI and was observed only for caspofungin against C. parapsilosis (4%, 1/23). As determined in time-kill studies, a caspofungin paradoxical effect was demonstrated by C. albicans (2/3), C. glabrata (1/3), and C. parapsilosis (1/3), including BSI that were determined to be negative by microdilution. In 50% human serum, a paradoxical effect was eliminated at caspofungin concentrations up to 64 μg/ml for 100% (8/8) of the C. albicans BSI. A caspofungin paradoxical effect was also eliminated by chitin synthase inhibitor nikkomycin Z and at achievable concentrations of calcineurin pathway inhibitors, tacrolimus and cyclosporine. Moreover, these agents were synergistic with caspofungin against 100, 100, and 88% (7/8) of C. albicans, respectively, and exerted their own paradoxical effects. Finally, paradoxical growth was eliminated in C. albicans irs4- and inp51-null mutants, which lack phosphatidylinositol-(4,5)-bisphosphate 5'-phosphatase. Our findings suggest that the paradoxical effect is unlikely to be important in vivo but remains an important tool to study cell wall stress responses. We implicate the Irs4-Inp51 phosphatidylinositol-(4,5)-bisphosphate 5'-phosphatase as a novel regulator of paradoxical growth.     

Correction: Anti-ulcerogenic effect of KFP-H008 against ethanol-induced gastric ulcer via p38 MAPK/NF-κB pathway

Correction for ‘Anti-ulcerogenic effect of KFP-H008 against ethanol-induced gastric ulcer via p38 MAPK/NF-κB pathway’ by Mei Su et al., RSC Adv., 2017, 7, 49423–49435.

The authors regret that the email address was not included for one of the three corresponding authors, Guang-ji Wang. Contact email addresses for all three corresponding authors are presented here.The Royal Society of Chemistry apologises for these errors and any consequent inconvenience to authors and readers.     

Activation of p38 MAPK in primary afferent neurons by noxious stimulation and its involvement in the development of thermal hyperalgesia

Alterations in the intracellular signal transduction pathway in primary afferents may contribute to pain hypersensitivity. We demonstrated that very rapid phosphorylation of p38 mitogen-activated protein kinase occurred in dorsal root ganglion (DRG) neurons that were participating in the transmission of noxious signals. Capsaicin injection induced phosphorylated-p38 (p-p38) in small-to-medium diameter sensory neurons with a peak at 2 min after capsaicin injection. Furthermore, we examined the p-p38 labeling in the DRG after noxious thermal stimuli and found a stimulus intensity-dependent increase in labeled cell size and the number of activated neurons. Most of these p-p38-immunoreactive (IR) neurons were small- and medium-sized neurons, which coexpressed transient receptor potential ion channel TRPV1 and phosphorylated-extracellular signal-regulated protein kinase. Intrathecal administration of the p38 inhibitor, FR167653, reversed the thermal hyperalgesia produced by the capsaicin injection. Inhibition of p38 activation was confirmed by the decrease in the number of p-p38-IR neurons in the DRG following capsaicin injection. Taken together, these findings suggest that the activation of p38 pathways in primary afferents by noxious stimulation in vivo may be, at least in part, correlated with functional activity, and further, involved in the development of thermal hyperalgesia.     

Selective inhibitors of MEK1/ERK44/42 and p38 mitogen-activated protein kinases potentiate apoptosis induction by sulindac sulfide in human colon carcinoma cells

The nonsteroidal anti-inflammatory drug (NSAID) sulindac prevents experimental colon cancer and can regress precancerous polyps in humans. Sulindac sulfide inhibits cyclooxygenase (COX)-mediated prostaglandin synthesis and retards the growth of cultured colon cell lines primarily by inducing apoptosis. Given the known role of mitogen-activated protein kinase (MAPK) in signal transduction and the regulation of cell survival and death, we determined the effect of sulindac sulfide on MAPK activation, COX-2 expression, and apoptosis induction in HCA-7 human colon cancer cells. Sulindac sulfide treatment was associated with activation of ERKp44/42 and p38 MAPK in a dosage- and time-dependent manner, and also activated upstream MEK. Similar results were seen in HCT-15 cells and also with the selective COX-2 inhibitor NS398. ERKp44/42 and p38 activation were accompanied by an induction of COX-2 protein expression. Selective inhibitors of sulindac sulfide-induced ERKp44/42 (PD98059) and p38 MAPK (SB203580) activation also suppressed the induction of COX-2 by this NSAID. Furthermore, both MAPK inhibitors significantly augmented sulindac sulfide-induced apoptosis, as did suppression of constitutive COX-2 using antisense oligonucleotides. In conclusion, MEK/ERK and p38 MAPK activation mediate COX-2 induction by sulindac sulfide. Selective inhibitors of these MAPKs potentiate apoptosis induction by this NSAID, suggesting a novel strategy for the prevention or treatment of colorectal cancer.     

全身炎症反应综合征与多器官功能障碍综合征患者外周血单核细胞COX-2与p38MAPK的关系

目的了解环氧化酶-2（COX-2）与p38丝裂原活化蛋白激酶（p38MAPK）在全身炎症反应综合征（SIRS）与多器官功能障碍综合征（MODS）患者发病机制中的作用。方法选择2004年6月-2005年6月广州中山大学属第一医院EICU、SICU与急诊观察区患者符合美国胸科医师协会／危重病医学会（ACCP／SCCM）SIRS和MODS诊断标准的患者28例,其中SIRS13例,MODS15例;与患者年龄、性别相区配的健康正常对照组11例。采用淋巴细胞分离液密度梯度离心法分离外周血单个核细胞（PBMCs）。用酶联免疫测定法（EIA）检测PBMCs的COX-2与p38MAPK含量;用逆转录．聚合酶链反应（RT-PCR）法检测PBMCs中COX-2与p38MAPK的mRNA的表达。用方差分析和简单相关分析法进行统计学分析。结果MODS组患者PBMCs的COX-2与p38MAPK含量及两者的mRNA表达均显著高于正常对照组和SIRS组（P均〈0．05）,正常对照组与SIRS组间差异无统计学意义;死亡患者高于存活患者（P〈0．05）;三组PBMCs中COX-2与p38MAPK含量呈正相关关系（r=0．663,P〈0．01）。死亡患者的外周血白细胞、淋巴细胞、氧合指数与存活患者比较差异无统计学意义;血糖、血肌酐则明显高于存活患者（P〈0．05）;二氧化碳总含量（TCO2）、pH明显低于存活患者（P〈0．01）。结论COX-2与p38MAPK参与了MODS的发病过程,而且可作为判断SIRS与MODS预后的参考指标之一。     

Rapamycin and UCN-01 synergistically induce apoptosis in human leukemia cells through a process that is regulated by the Raf-1/MEK/ERK, Akt, and JNK signal transduction pathways

Interactions between the protein kinase C and Chk1 inhibitor UCN-01 and rapamycin in human leukemia cells have been investigated in relation to apoptosis induction. Treatment of U937 monocytic leukemia cells with rapamycin (10 nmol/L) in conjunction with a minimally toxic concentration of UCN-01 (100 nmol/L) for 36 hours resulted in marked potentiation of mitochondrial injury (i.e., loss of mitochondrial membrane potential and cytosolic release of cytochrome c, AIF, and Smac/DIABLO), caspase activation, and apoptosis. The release of cytochrome c, AIF, and Smac/DIABLO were inhibited by BOC-D-fmk, indicating that their release was caspase dependent. These events were associated with marked down-regulation of Raf-1, MEK, and ERK phosphorylation, diminished Akt activation, and enhanced phosphorylation of c-Jun NH2-terminal kinase (JNK). Coadministration of UCN-01 and rapamycin reduced the expression levels of the antiapoptotic members of the Bcl-2 family Mcl-1 and Bcl-xL and diminished the expression of cyclin D1 and p34(cdc2). Furthermore, enforced expression of a constitutively active MEK1 or, to a lesser extent, myristoylated Akt construct partially but significantly attenuated UCN-01/rapamycin-mediated lethality in both U937 and Jurkat cell systems. Finally, inhibition of the stress-related JNK by SP600125 or by the expression of a dominant-negative mutant of c-Jun significantly attenuated apoptosis induced by rapamycin/UCN-01. Together, these findings indicate that the mammalian target of rapamycin inhibitor potentiates UCN-01 cytotoxicity in a variety of human leukemia cell types and suggest that inhibition of both Raf-1/MEK/ERK and Akt cytoprotective signaling pathways as well as JNK activation contribute to this phenomenon.