82倍和2.10倍(P
目的了解CdCl2对腺垂体的损伤以及细胞凋亡发生与p38MAPK、ERK12表达的关系,为了解镉致腺垂体毒作用的分子机制提供科学依据。方法采用健康雄性SD大鼠进行整体试验(n=12),分别每天经口灌胃给予0、1.0、2.0、4.0mgkgbwCdCl2,6周后取腺垂体进行指标检测;离体试验采用酶解分离大鼠之原代腺垂体细胞,分别以0、1.56、3.12、6.25、12.50、25.00、50.00、100.00μmolLCdCl2处理,收获细胞进行检测;特异性阻断剂阻断凋亡效应的试验采用2.65μmolLp38MAPK的特异阻断剂SB203580或10μmolLERK12激酶的特异阻断剂U0126处理细胞,然后以3.12μmolL或100.00μmolL的CdCl2处理细胞后进行相应的检测。检测指标包括:TUNEL法和流式细胞术等检测凋亡。结果整体实验和离体实验结果均显示CdCl2以剂量依赖方式诱导腺垂体细胞发生凋亡(P<0.05);特异性阻断剂效应的研究结果显示2.65μmolLSB203580和10μmolLU0126对TUNEL阳性细胞的相对灰度和凋亡细胞率均有一定的影响。结论在一定的剂量条件下,CdCl2影响腺垂体激素分泌水平,并导致发腺垂体细胞凋亡,MAPKs家族成员p38MAPK和ERK12激酶通路在凋亡发生过程中可能发挥一定的作用。
目的:观察manumycin对人胰腺导管癌细胞Panc-1的抑制效应,并探讨其诱导细胞凋亡是否经
BGB324小白鼠 p38MAPK介导。方法:用MTT法检测manumycin对Panc-1细胞的抑癌作用。用caspase-3活性检测试剂盒定量检测 manumycin诱导细胞凋亡的水平及评估特异性的p38MAPK抑制剂SB203580对它的影响。结果:经manumycin Bortezomib制造商 (6／μmol／L、18／μmol／L、54 μmol／L)处理Panc-1细胞24 h,对Panc-1细胞生长具有明显的抑制作用,其抑制率分别为 8．9％、21．9％和67．0％,其中后二者的细胞活性与对照组相比有显著差异(P
目的:通过瞬时转染p38MAPK途径中上游激酶:组成激活型MAPK激酶3(MKK3b)和MAPK激酶6(MKK6b),进一步了解p38MAPK级联传导信号系统调节iNOS基因在胶质细胞中的转录激活机制。方法:MKK3b或MKK6b与接有荧光素酶(luciferase,Luc)的大鼠iNOS启动基因质粒(iNOS-Luc);cAMP反应元件(CRE-Luc)和核因子κB(NFκB-Luc)联合转染C6胶质细胞株。结果:MKK3b/MKK6b能引起iNOS-Luc的激活,并都能够被p38MAPK抑制剂SB203580所抑制。MKK可以诱导cAMP反应元件(CRE-Luc)介导的和核因子κB(NFκB-Luc)依赖的转录活性。显性抑制型(dominantnegative,dn)CRE结合蛋白(dnCREB)和CCAAT/增强子结合蛋白(C/EBP)都是p38MAPK的靶向作用目标。结论:转染这两种转录因子产生相反的影响:dnCRE增强了iNOS-Luc的表达,而dnC/EBP则引起抑制作用,对CRE-Luc也具有相同的影响。
Aim:
Interleukin-1 beta (IL-1β) has been implicated as an extracellular signal in the initiation of apoptosis in neurons and oligodendrocytes after spinal cord injury (SCI). To further characterize the apoptotic cascade initiated by IL- 1β after SCI, we examined the expression of IL-1β, p38 mitogen-activated protein kinase (p38 MAPK) and caspase-3 after SCI, and further investigated whether p38 MAPK was involved in neuron apoptosis induced by IL- 1β. Methods: Adult rats were given contusion SCI at the T-10 vertebrae level with CX-4945化学结构 a weight-drop impactor (10 g weight dropped 25.0 mm). The expression levels of IL-1β, p38 MAPK and caspase-3 after SCI were assessed with Western blots, immunohistochemistry staining, and real time reverse transcription polymerase chain reactions (RT-PCR). Neuron apoptosis was assessed with the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-biotin nick end labeling (TUNEL) method. Results: Increased levels of IL-1β and p38 MAPK were observed soon after injury, with a peak in expression levels within 6 h of injury. By 24 h after injury, caspase-3 expression was markedly increased in the injured spinal cord.