脂多糖激活p38在诱导肿瘤坏死因子α基因表达中的作用

目的探讨防治内毒素休克的新方法,研究脂多糖（ＬＰＳ）诱导肿瘤坏死因子α（ＴＮＦα）表达的分子机制。方法用蛋白激酶活性测定检测ＬＰＳ刺激引起的激酶活性变化;共聚焦激光扫描技术显示ｐ３８激活移位;用反转录聚合酶链反应和报告基因系统研究ＴＮＦα基因转录的分子机制。结果发现ＬＰＳ刺激ＲＡＷ细胞引起ｐ３８激活并由胞浆移位至胞核。ＬＰＳ刺激引起ＴＮＦαｍＲＮＡ表达增加,而且由ＬＰＳ引起的ＴＮＦα的转录活性可被ｐ３８特异性抑制剂所抑制。结论激活的ｐ３８通过磷酸化转录因子增加ＴＮＦα基因转录活性,这是中毒性休克时ＴＮＦα生成增加的一个重要机制。ｐ３８是ＬＰＳ诱导ＴＮＦα基因表达的重要调节物质。     

丝裂原活化蛋白激酶通路对脂多糖诱导RAW264.7细胞肿瘤坏死因子α基因表达的协同调节作用

目的 探讨脂多糖（LPS）诱导RAW264.7细胞肿瘤坏死因子α(TNF-α)基因表达过程中丝裂原活化蛋白激酶（MAPK）通路的协同调节作用及其分子机制。方法 用蛋白激酶活性测定分析LPS刺激RAW264．7细胞引起的激酶活性变化;用报告基因技术和反转录聚合酶链反应（RT－PCR）方法研究LPS诱导的TNF－α基因转录的分子机制。结果 LPS刺激RAW264．7细胞可引起细胞外信号调节激酶1（ERK1）、c-Jun氨基末端激酶1（JNK1）和p38MAPK的一过性激活,用MAPK上游激酶的活性突变体分别转染RAW264．7均可不同程度地诱导TNF－α启动子转录活性;而且,这些MAPK通路激活诱导的TNF－α启动子转录活性表现出明显的协同效应;三种MPAPK的无活性突变体均显示出对LPS刺激引起的TNF－α启动子转录激活的抑制效应;RT－PCR的结果证实,ERK、JNK和p38MAPK的特异性抑制剂对TNF－αmRNA表达具有不同程度的抑制作用。结论 LPS刺激引起的TNF－α启动子转录活性增加,可能涉及了ERK、p38和JNK三条通路的激活;这些通路通过协同效应共同发挥对TNF－α基因表达的调控。     

LPS刺激对RAW264.7细胞p38蛋白激酶的细胞内定位的影响

目的 探讨p38蛋白激酶信号传导的过程及其在细胞中的特异性作用机制。方法 应用间接荧光标记免疫探针技术及共聚焦激光扫描技术观察单核细胞中p38蛋白激酶的分布及LPS对其分布的影响。结果p38在未受刺激的静息单核细胞及内皮生长因子（EGF）刺激的单核细胞胞浆和胞核中荧光强度均呈弥散性分布;脂多糖（LPS）刺激使细胞核区的荧光强度明显增强,而胞浆区域的荧光强度降低。激酶动力学的研究显示,p38激活先于p38移位。LPS刺激后30 min,p38激酶活性即达到高峰,随后2 h内逐步下降,p38激酶活性呈一过性增高;LPS刺激45 min后,核区荧光强度达到峰值,且在2 h内维持在高水平。结论 单核细胞由LPS激活后,其p38蛋白激酶由胞浆转位到胞核,反应具有一定的特异性;p38移位入核依赖于p38磷酸化活化及由细胞浆移位到细胞核是一系列连续发生的事件。     

p38丝裂原激活蛋白激酶对人胚胎肾293细胞一氧化氮合酶基因转录的调控

目的：探讨p38丝裂原激活蛋白激酶（MAPK）家族四种亚型对诱导型一氧化氮合酶（iNOS)基因的转录调控。方法：以人胚胎肾293（HEK293）细胞为靶细胞，采用脂质体（LPS）介导的细胞基因共转染技术，荧光素酶报告基因技术，分别将FLAG－tagged p38MAPK4种亚型，含有鼠iNOS基因启动子区的荧光素酶报告基因质粒（piNOS-Luc)，空载体（pcDNA3),β－半乳糖苷酶表面质粒（pCMV－β）共转染，检测并比较荧光素酶相对活性。结果（1）未加刺激时，在HEK293细胞中，p38MAPK中仅有p38α的诱导作用亦有明显，结论：（1）LPS能够在HEK293细胞中诱导iNOS基因转录活性；（2）在HEK293细胞中，p38MAPK参与了静息时及LPS刺激下对iNOS基因的转录调控。     

Biochemical pathways in the antiatherosclerotic effect of berberine

Background This study investigated the inhibitory effect of berberine （BBR） on lipopolysaccharide （LPS） induced cyclooxygenase-2 （COX-2） expression via the mitogen activated protein kinase （MAPK） signalling cascade pathways in human peripheral blood monocytes （PBMC）.
Methods PBMC from whole blood were isolated and cultured for up to 24 hours after division into 5 groups treated with LPS, LPS＋BBR 25 μmol/L, LPS＋BBR 50 μmol/L or LPS＋BBR 100 μmol/L and untreated. Monocytes were extracted for RT-PCR and Western blot analyses to examine COX-2 mRNA and protein activated expression of p38 mitogen activated protein kinase （p38MAPK）, Jun N-terminal kinase （JNK） and extracellular regulated kinases 1/2 （ERK1/2） signalling pathways.
Results COX-2 mRNA and protein expression decreased to a minimum at 12 hours after BBR treatment （P 〈0.05）. With the increasing concentration of BBR treatment, the COX-2 expression decreased progressively （P 〈0.01）. With BBR treatment for 6, 12 or 24 hours at three doses, ERK1/2 protein expression was significantly inhibited. For the JNK pathway, only with the treatment of BBR at the concentration of 100 μmol/L was JNK protein expression inhibited compared with the LPS stimulation group （P 〈0.01）. Irrespective of the BBR concentration, no difference was shown between the BBR group and the LPS group for p38MAPK protein expression. Human monocytes COX-2 mRNA, by RT-PCR, and protein expression, by Western blot analysis, were inhibited when incubated with PD98059, SP600125 and SB203580 （P 〈0.05）.
Conclusions Berberine inhibits COX-2 expression via the ERK1/2 signalling pathway and, possibly, at a high dosage via the JNK pathway. P38MAPK may have no relationship with the effect of BBR in PBMC. Berberine inhibited COX-2 mRNA and protein expression in a dose dependent manner and suppressed COX-2 expression to a minimal level after 12 hours of berberine treatment.     

Studies on the translocation of p38 mitogen-activated protein kinase in cardiomyocytes of nude mice on the stimulation of LPS

Milogen-activate(l I)rotejn kinase (MAPK) is the major mediator to lransduce the extracellular signals from tilem(fll]1)rarle to internal nucleus"l. p38 MAPK pathway isone of 1\he 4 pathways that have been identified sofar',.,"'. l[ was proved that the aotivati(,n of I)38 MAPKall'e(fled Ills I)r(>(t'fsses of (tell growth, cell 'lycle and aPOptosis. afl(1 was involve(l ifl the uvula[1()n of inflammationan(l sir(>ss l-c):il)onses. Tile sin(ly ')f the exa(II (lellular location oj' I)n)lo…     

丝裂原活化蛋白激酶对佛波酯诱导滋养细胞MMP-9基因表达的影响

目的 探讨佛波酯(PMA)诱导细胞滋养层细胞(CTB)MMP-9基因表达的调控机制。方法 用细胞ELISA法测定CTB细胞的蛋白激酶活性变化；用反转录聚合酶链反应检测CTB中MMP-9的基因表达。结果 100nmol／L PMA能迅速激活CTB中丝裂原活化蛋白激酶(MAPK)家族中细胞外信号调节蛋白激酶(ERK)、c-jun氨基末端激酶(JNK)以及p38 MAPK激酶的活性。100nmol／L PMA刺激CTB引起MMP-9 mRNA表达显著增加，能被ERK或p38 MAPK的特异性抑制剂所抑制。结论 ERK和p38 MAPK可能是PMA诱导CTB中MMP-9基因表达增加的重要调节物质。     

丝裂原活化蛋白激酶对佛波酯诱导滋养细胞MMP-9基因表达的影响

目的 探讨佛波酯(PMA)诱导细胞滋养层细胞(CTB)MMP-9基因表达的调控机制。方法 用细胞ELISA法测定CTB细胞的蛋白激酶活性变化;用反转录聚合酶链反应检测CTB中MMP-9的基因表达。结果 100 nmol/L PMA能迅速激活CTB中丝裂原活化蛋白激酶(MAPK)家族中细胞外信号调节蛋白激酶(ERK)、c-jun氨基末端激酶(JNK)以及p38 MAPK激酶的活性。100 nmol/L PMA刺激CTB引起MMP-9 mRNA表达显著增加,能被ERK或p38 MAPK的特异性抑制剂所抑制。结论 ERK和p38 MAPK可能是PMA诱导CTB中MMP-9基因表达增加的重要调节物质。     

脂多糖通过p38/MAPK通路对巨噬细胞自噬的调控作用

摘 要:【目的】 观察脂多糖(LPS)对巨噬细胞自噬的影响及p38/MAPK通路在其中的作用?【方法】 体外培养巨噬细胞株RAW264.7,分为对照组?饥饿状态激活自噬组?单纯LPS刺激组?LPS+P38抑制剂(SB203582)组和LPS+mTOR抑制剂(rapamycin)组?将前期构建的载体pcDNA3.1-GFP-LC3,转染RAW264.7,通过荧光显微镜观察各组细胞中自噬体形成情况?qRT-PCR方法检测各组中与细胞自噬相关的基因Atg5,Atg7,LC3-Ⅱ和Bnip3 mRNA表达水平的改变?利用Western blotting检测LC3-Ⅱ?p-P38?P38蛋白在各组中的表达情况,以评价LPS激活巨噬细胞自噬过程中p38/MAPK通路的作用?【结果】 在荧光显微镜下可以观察到自噬在饥饿状态组?LPS+SB203582组和LPS+rapamycin组有明显增强;qRT-PCR检测到自噬相关基因Atg5,Atg7,LC3-Ⅱ,和Bnip3 mRNA的表达在饥饿状态组?LPS+SB203582组和LPS+rapamycin组有明显增强;Western blotting 检测发现p-P38在饥饿状态组?LPS组和LPS+rapamycin组中表达明显升高; LC3-Ⅱ的表达在饥饿状态组?LPS+SB203582组和LPS+rapamycin组中表达要高于对照组和LPS组?【结论】 LPS参与巨噬细胞自噬的调控,除经典mTOR通路之外,p38/MAPK通路是其抑制通路之一?     

p38MAPK在脂多糖诱导的人内皮细胞表达诱导型一氧化氮合酶中的作用

目的：研究P38丝裂原激活蛋白激酶（P38MAPK）在脂多糖（LPS）诱导的人内皮细胞－ECV304诱导一氧化氮合酶（iNOS)表达的一氧化氮（NO）产生中的作用，方法：用Griess法检测人内皮细胞培养液中NO水平，分别用免疫荧光法和RT－PCR检测细胞iNOS蛋白和mRNA的表达，采用免疫沉淀法检测细胞p38MAPK的活性，结果：与对照组相比，LPS刺激后的ECV304细胞iNOS蛋白和mRNA的表达显著增加，用p38特异性抑制剂SB2023580预处理后，可以显著抑制细胞iNOS的表达和NO的产生，LPS 刺激内皮细胞后15min,p38MAPK的活性达到高峰，维持45min后逐渐下降，结论：p38MAPK参与与LPS诱导的内皮细胞iNOS的表达和NO的产生，可通过阻断信号转导通路来减少iNOS及其他细胞因子的产生，为败血症休克的防治提供一条新的思路。     

UV通过p38MAPK促进大鼠胶质瘤C6细胞凋亡

目的：探讨p38MAPK在紫外线损伤刺激细胞中的特异性信号转导作用。方法：流式细胞仪检测紫外线照射30min后1，2及4h的C6细胞周期变化和是否有凋亡发生；应用免疫细胞化学技术观察紫外线刺激前后p38MAPK在C6细胞中的表达强度和分布特征。结果：细胞周期结果显示1，2和4h后C1期细胞数分数各增多0．12，0．21和0．19，而S期细胞数分数减少0．10，0．14和0．15；各组的凋亡率分别是12％，49％和34％；未受刺激的细胞中，p38MAPK在胞质和胞核表达较弱；紫外线损伤作用2h后，细胞核区的染色强度即明显增强，而胞质区域的染色强度相对降低。结论：C6细胞受紫外线损伤后可通过p38MAPK通路发生凋亡。     

p38 MAPK在大鼠急性肺损伤模型中的表达及抗氧化剂NAC的影响

目的:观察磷酸化的分裂原激活的蛋白激酶(p38 MAPK)在脂多糖(LPS)引起的大鼠急性肺损伤中在体原位的表达. 方法:采用免疫组织化学ABC法和Western-blot技术,检测磷酸化的p38 MAPK在大鼠急性肺损伤模型气道和肺组织的表达,并同时观察N-乙酰半胱氨酸(NAC)对其表达的影响. 结果:对照组气道和肺组织无或偶见反应极弱的p38 MAPK阳性细胞,散在分布于气道上皮细胞和肺泡上皮细胞. LPS致伤组p38 MAPK阳性细胞较对照组明显增多(P<0.01),主要分布于浸润的炎症细胞、气道上皮细胞、肺泡上皮细胞、胸膜间皮细胞和血管内皮细胞. NAC治疗组气道和肺组织中阳性细胞数较LPS致伤组明显减少(P<0.01),Western-blot结果验证了上述结果. 结论:LPS诱发的大鼠急性肺损伤模型中,磷酸化p38 MAPK在气道和肺组织内表达增加,p38 MAPK的激活见于肺组织内多数细胞,提示肺内炎性和非炎性细胞均有p38 MAPK信号分子的激活. NAC是有效的抗氧化剂,其对急性肺损伤的炎症抑制可能是通过抑制p38 MAPK的激活起作用.     

p38 MAPK在脂多糖诱导的人内皮细胞表达诱导型一氧化氮合酶中的作用

目的研究p38丝裂原激活蛋白激酶(p38 MAPK)在脂多糖(LPS)诱导的人内皮细胞－ECV304诱导型一氧化氮合酶(iNOS)表达和一氧化氮(NO)产生中的作用。方法用Griess法检测人内皮细胞培养液中NO水平,分别用免疫荧光法和 RT－PCR检测细胞 iNOS蛋白和mRNA的表达,采用免疫沉淀法检测细胞 p38 MAPK的活性。结果 与对照组相比,LPS刺激后的BCV304细胞iNOS蛋白和mRNA的表达显著增加,用p38特异性抑制剂SB203580预处理后,可以显著抑制细胞 iNOS的表达和NO的产生。LPS刺激内皮细胞后15 min,p38 MAPK的活性达到高峰,维持45 min后逐渐下降。结论p38MAPK参与了LPS诱导的内皮细胞iNOS的表达和NO的产生;可通过阻断信号转导通路来减少iNOS及其他细胞因子的产生,为败血症休克的防治提供一条新的思路。     

Effect of cigarette smoke extract on lipopolysaccha- ride-activated mitogen-activated protein kinase signal transduction pathway in cultured cells

Background  Lipopolysaccharide (LPS) forms outer membrane of the wall of Gram-negative cells. LPS can directly cause damage to epithelia of respiratory tract and is the major factor responsible for the chronic inflammation of respiratory passage. The mitogen-activated protein kinase (MAPK) signal transduction pathway of the airway epithelia is intimately associated with the action of LPS. The chronic inflammation of respiratory tract and smoking are interrelated and entwined in the development and progression of chronic lung diseases. This study was designed to examine the effects of cigarette smoke extract (CSE) and LPS on MAPK signal transduction pathway in order to further understand the roles CSE and LPS play in chronic lung inflammation.
Methods  Cultured primary human epithelial cells of airway were divided into four groups according to the stimulants used: blank control group, LPS-stimulation group, CSE-stimulation group and CSE plus LPS group. Western blotting was employed for the detection of phosphorylation level of extracellular-signal-regulated-kinase (ERK_1/2), p38 MAPK and c-Jun N-terminal kinase (JNK). The expression of cytokines of MAPK transduction pathway (granulocyte-macrophage colony stimulating factor (GM-CSF) and mRNA of IL-8) in the primary epithelial cells of respiratory tract was also determined.
Results  Western blotting revealed that the phosphorylation levels of ERK_1/2, p38 MAPK and JNK were low and 2 hours after the LPS stimulation, the phosphorylation of ERK_1/2, p38 MAPK and JNK were all increased. There was a significant difference in the phosphorylation between the LPS-stimulation group and blank control group (P<0.05); no significant difference was found between CSE-stimulation group and blank control group (P>0.05); there was a significant difference between CSE + LPS group and blank control group and between CSE + LPS group and LPS group (P<0.05). The phosphorylation of CSE-LPS group was higher than that of blank control group but lower than that of LPS group. In blank control group, the expression of IL-8 and GM-CSF mRNA was low in the epithelial cells of airway and the release of IL-8 and GM-CSF was also at a low level. One hour after LPS stimulation, the level of IL-8 mRNA increased (P<0.05) and reached a peak after 2 hours. On the other hand, GM-CSF mRNA level increased 2 hours after the stimulation (P<0.05) and reached the highest level 4 hours after the stimulation. Two hours after LPS stimulation, IL-8 and GM-CSF protein level began to rise (P<0.05), and the level was the highest 8 hours after the stimulation (P<0.01). Stimulation with CSE alone had no effect on the release of IL-8 and GM-CSF and expression of IL-8 mRNA (P>0.05), but pre-treatment with CSE could delay the LPS-induced release of IL-8 and GM-CSF and the expression of IL-8 mRNA and its peak was lower.
Conclusions  LPS stimulation can significantly increase the phosphorylation of ERK_1/2, p38 MAPK and JNK in the epithelial cells of airway and activate the MAPK transduction pathway, thereby can activate the downstream signal transduction pathway, and can ultimately result in the release of cytokines by the epithelial cells of airway. CSE can partially abolish the LPS-induced activation of MAPK signal transduction pathway and the expression of cytokines of the pathway, which might contribute to the development and progression of the inflammatory reactions in COPD patients.

     

Simvastatin attenuates lipopolysaccharide-induced airway mucus hypersecretion in rats

Background Mucus hypersecretion in the respiratory tract and goblet cell metaplasia in the airway epithelium contribute to the morbidity and mortality associated with airway inflammatory diseases. This study aimed to examine the effect and mechanisms of simvastatin on airway mucus hypersecretion in rats treated with lipopolysaccharide （LPS）. Methods Mucus hypersecretion in rat airways was induced by intra-tracheal instillation of LPS. Rats treated with or without LPS were administered intra-peritoneally simvastatin （5 and 20 mg/kg） for 4 days. Expression of Muc5ac, RhoA and mitogen-activated protein kinases （MAPK） p38 in lung were detected by real-time polymerase chain reaction （PCR）, immunohistochemistry or Western blotting. Tumor necrosis factor （TNF）-α and IL-8 in bronchoalveolar lavage fluid （BALF） were assayed by an enzyme-linked lectin assay and enzyme linked immunosorbent assay （ELISA）. Results Simvastatin attenuated LPS-induced goblet cell hyperplasia in bronchial epithelium and Muc5ac hypersecretion at both the gene and protein levels in lung （P 〈0.05）. Moreover, simvastatin inhibited neutrophil accumulation and the increased concentration of TNF-α and IL-8 in BALF follows LPS stimulation （P 〈0.05）. The higher dose of simvastatin was associated with a more significant reduction in Muc5ac mRNA expression, neutrophil accumulation and inflammatory cytokine release. Simultaneously, the increased expression of RhoA and p38 MAPK were observed in LPS-treated lung （P 〈0.05）. Simvastatin inhibited the expression of RhoA and p38 phosphorylation in lung following LPS stimulation （P 〈0.05）. However, the increased expression of p38 protein in LPS-treated lung was not affected by simvastatin administration. Conclusions Simvastatin attenuates airway mucus hypersecretion and pulmonary inflammatory damage induced by LPS. The inhibitory effect of simvastatin on airway mucus hypersecretion may be through, at least in part, the suppression of neutrophil accumulation and     

Mechanism of reduction of albumin expression induced by lipopolysaccharide in rat hepatocytes

Background The severity of hypoalbuminemia has been shown to be related to morbidity and mortality in some critical illnesses, illustrating the need for better understanding of molecular mechanism of hypoalbuminemia. Lipopolysaccharide (LPS) is a key mediator inducing hypoalbuminemia in sepsis and septic shock. The present study was designed to identify if the reduction of albumin expression is directly induced by LPS and modulated by activated extracellular signal-regulated protein kinase (ERK) and p38 mitogen-activated protein kinase (MAPK) in rat hepatocytes.Methods Primary rat hepatocytes were divided into five groups. In two of them, hepatocytes were treated with normal saline or 1 μg/ml LPS, then albumin mRNA expression was observed at 0, 2, 8, 12 and 24 hours after treatment. In another group, hepatocytes were pretreated with 100, 40 or 20 μmol/L of cycloheximide (CHX, an inhibitor of protein synthesis) for 30 minutes followed by 1 μg/ml LPS for 24 hours. Then the RNA was extracted from the cells for RT-PCR to detect the expression of albumin. The other two groups were administered 1 μmol/L, 10 μmol/L and 50 μmol/L of SB203580 (p38 MAPK inhibitor) or PD98059 (ERK inhibitor) 30 minutes prior to 1 μg/ml LPS treatment. After 24 hours of LPS treatment, the supernatant was collected and assayed for albumin concentrations. Data were analyzed by one-way analysis of variance, followed by the Newman-Keul test; a P&lt;0.05 was considered significant.Results There was no marked change in albumin mRNA expression in the control group during 24-hours treatment with normal saline. The reduction did not occur until 24 hours after LPS treatment, and albumin mRNA decreased by 30% approximately compared to the control group at 24 hours (0.587 vs 0.832, P=0.007). CHX could inhibit the decline of albumin mRNA induced by LPS and the effect was correlated with the dose of CHX. The ERK inhibitor PD98059 caused a significant increase in LPS-induced albumin production at the three concentrations (119.7, 111.4 and 80.0 ng/ml vs 44.4 ng/ml, P=0.0013, 0.0025 and 0.009, respectively), whereas SB203580 obviously blocked albumin reduction in LPS-treated cells at the concentrations of 10 and 50 μmol/L (87.5 and 93.6 ng/ml vs 44.4 ng/ml, P=0.0076 and 0.0049, respectively).Conclusions LPS can induce the reduction of albumin expression by new synthesized proteins indirectly, and the process may be related to the signal proteins of ERK and p38 kinase. The ERK and p38 kinase are critical signaling pathways in LPS-induced hypoalbuminemia which is worthwhile to understand in studying the molecular mechanism of hypoalbuminemia in sepsis and septic shock.     

survivin反义寡核苷酸诱导人肝癌细胞凋亡的相关信号转导途径

目的 利用人肝癌细胞株SMMC-7721细胞,探讨survivin基因诱导肝癌细胞凋亡的相关信号转导途径。方法 采用脂质体介导survivin反义寡核苷酸转染人肝癌细胞株SMMC-7721细胞。采用Western印迹及原位杂交检测细胞内survivin蛋白及mRNA的表达。采用流式细胞仪及TUNEL染色检测细胞凋亡的变化。采用western印迹,免疫沉淀,逆转录-聚合酶链反应（RT-PCR）及激酶活性检测方法测定细胞内丝裂原活化蛋白激酶p38(p38MAPK)及半胱氨蛋白水解酶3表达及活性的变化,观察p38MAPK与半胱氨蛋白水解酶3活性变化之间的关系。结果 脂质体介导转染survivin反义寡核苷酸后,SMMC-7721肝癌细胞内survivin蛋白及mRNA表达可分别被下调84.6%及69.7%。凋亡细胞比率由转染前的0.70%增加至31.51%。转染后细胞内p38MAPK及半胱氨蛋白水解酶3表达无明显变化。但其活性显著升高,其中p38MAPK活性升高发生于半胱氨蛋白水解酶3活性升高之前。结论 转染survivin反义寡核苷酸可以有效诱导肝癌细胞发生凋亡,其诱导肝癌细胞凋亡的信号通过顺序激活p38MAPK-半胱氨蛋白水解酶3信号途径传导。     

p38MAPK通路在佛波酯诱导人绒癌JAR细胞体外侵袭中的作用

目的 研究细胞内p38MAPK信号传导通路在人绒癌JAR细胞体外侵袭中的作用。方法 用ELISA法测定JAR细胞中p38MAPK的活性变化；用Tramwell细胞侵入系统检测细胞的侵袭作用；用MTT法评价细胞生长状况。结果 佛波酯(PMA)呈浓度依赖性地激活JAR细胞中p38MAPK。PMA能促进人绒癌JAR细胞的体外侵袭作用，而p38特异性抑制剂SB203580抑制了JAR细胞的侵袭能力。结论 p38MAPK通路在人滋养细胞的侵袭行为以及人绒癌的形成中具有重要作用。p38MAPK抑制剂可能会为人绒癌的防治提供新的途径。     

己酮可可碱对p38丝裂原激活蛋白激酶激活的影响

目的:观察己酮可可碱(PTX)对p38丝裂原激活蛋白激酶(MAPK)激活的影响,探讨PTX保护内皮细胞单层通透性的机制.方法:建立内皮细胞单层损伤模型,随机分为脂多糖(LPS)-A组、LPS-B组、LPS PTX-A组及LPS PTX-B组,分别以不同终浓度LPS及PTX刺激不同时间,以Western印迹杂交技术检测内皮细胞p38MAPK活性.结果:LPS组,随着LPS刺激时间的延长及刺激浓度的增大,磷酸化的p38MAPK亮度逐渐增强;LPS PTX组,磷酸化的p38MAPK亮度随着PTX刺激时间的延长及刺激浓度的增大而逐渐减弱.结论:PTX可通过降低p38 MAPK的活性而起保护内皮细胞通透性的作用.