应用基因芯片研究代谢相关基因在先兆子痫胎盘中的表达

目的:研究部分代谢相关基因在先兆子痫胎盘组织中的表达变化。方法:采用包含220余种人类环境激素相关基因cDNA片段的基因芯片,对经过严格配伍的正常和先兆子痫胎盘组织中基因表达谱的差异进行检测。结果:先兆子痫胎盘中与氧化还原代谢相关的基因(GenBank:U78168、X16699、D32143、AL035079、AF069668、X53463、J03746、X02317、X91247等)表达增高。另外,与激素等其它代谢相关的一些基因(如GenBank:NM_-001718、Z22535、M38180、M76665、X75252、J03258、U56725)在先兆子痫胎盘中的表达也增强。结论:胎盘内基因表达的变化与先兆子痫患者体内多种代谢改变有关。     

妊娠高血压综合征胎盘中病理相关基因的表达改变

目的研究与细胞因子信号转导等相关的病理基因在妊娠高血压综合征(妊高征)胎盘组织中的表达变化.方法采用分别包含220余种人类细胞因子相关基因或人类环境激素相关基因cDNA片段的基因芯片,检测严格配伍的正常和妊高征胎盘组织中基因表达谱的差异.结果一些疾病相关基因(GenBank:U82828、X15183)在妊高征患者胎盘中的表达高于正常,另外一些与基因表达调控有关的转录因子基因(GenBank:AL022312、M15990、U15009、L49380等)也表达增强.此外,妊高征胎盘中与细胞因子信号转导有关的大多数受体/激酶表达增强,例如GenBank登录号为M76125、U73531、L76191、L06139、AF035121、M23379、U43195等基因.结论细胞因子信号转导相关基因等的表达增强提示细胞因子的功能活跃,进一步说明胎盘局部细胞因子水平的变化与妊高征的病理发生密切相关.     

滋养层细胞侵入相关基因在先兆子痫胎盘中的表达

探讨与滋养层侵入有关的细胞外基质分子相关基因在先兆子痫胎盘中的表达，采用分别点样有220余种人细胞因子相关基因和人类激素相关基因cDNA片段的两款基因芯片，检测经过严格配伍的先兆子痫和正常胎盘组织的基因表达谱差异。结果显示：钙粘蛋白、胶原、整合素、选择蛋白等18种细胞外基质分子基因的表达在先兆子痫和正常胎盘组织间相差2倍以上，且全部表现为在先兆子痫胎盘中的表达增强。先兆子痫患者的胎盘组织中基质金属蛋白酶(MMP)-10、-13、-15和金属蛋白酶组织抑制因子(TIMP)-2、TIMP-3、纤溶酶原、纤溶酶原激活物等的表达均较正常者高。提示胎盘中细胞外基质分子及其降解酶基因表达异常可能与先兆子痫的病理发生关系密切。     

扩张型心肌病细胞因子及糖皮质激素受体的研究

检测 30例扩张型心肌病 (DCM )外周血白细胞介素 1(IL 1)、白细胞介素 6 (IL 6 )及肿瘤坏死因子 (TNF α )的水平 ,并同时测定白细胞糖皮质激素受体 (GR )的水平 ,结果发现DCM患者的IL 1、IL 6及TNF α明显高于对照组 (P <0 0 1) ,而GR明显低于对照组 (P <0 0 1) ,表明DCM存在细胞因子及GR异常 ,并对其意义进行了分析。     

信号分子TRAF6的研究讲展

肿瘤坏死因子受体相关因子(tumor necrosis factor receptor-associated factors,TRAFs)是肿瘤坏死因子(tumor necrosis factor,TNF)超家族和Toll样/白细胞介素-1受体(TOIV IL-1 receptor,TIR)超家族重要的接头分子,在天然免疫和获得性免疫中发挥重要作用.     

先兆子痫胎盘的基因表达谱研究

目的： 利用cDNA芯片分析先兆子痫胎盘中基因表达的变化情况,寻找新的先兆子痫相关基因。方法: 建立含12 000个与代谢、凋亡、细胞黏附、信号转导、转录因子等有关基因的cDNA表达谱芯片,将先兆子痫及正常胎盘mRNA与cDNA芯片进行杂交,得到先兆子痫的基因表达谱;对部分差异表达基因进行Northern验证。结果: 在4例先兆子痫胎盘组织中发现均有差异表达的基因44个,其中表达上调有30个,表达下调有14个,Northern杂交鉴定的结果与芯片结果相符合。结论: 重度妊娠高血压疾病的基因表达谱存在明显差异,差异的基因可能涉及信号转导、细胞代谢、炎性细胞因子等方面,这些基因可能与妊娠高血压病的发病相关。     

利用抑制性减数杂交技术(SSH)研究IL-10抑制表达的新基因

目的　利用抑制性减数杂交 (SSH)技术筛选IL 10抑制表达的新基因。方法　以PHA刺激的U937细胞为tester,以PHA刺激同时IL 10抑制的U937细胞为driver,提取mRNA ,反转录构建cDNA文库 ,利用SSH技术筛选IL 10抑制表达的新基因。结果　通过SSH技术 ,发现了 15个可能的IL 10抑制表达的基因 ,其中 6个与GenBank中已公布的表达序列检签 (expressedsequencetag ,EST)片段没有明显同源性 ,为新基因 ,其中之一经EST拼接 ,得到全长cDNA ,为一种新的C C家族趋化因子 ,已被GenBank收录 ,收录号码为AF0 96 985。挑选 4个新基因进行Northernblot分析 ,发现IL 10可抑制其中 3个新基因表达。结论　SSH技术是一种高效的差异基因筛选方法 ,IL 10可抑制多种包括新细胞因子基因的表达     

代谢相关基因在先兆子痫胎盘中的表达情况分析

目的探讨细胞代谢相关基因在先兆子痫胎盘组织中的表达变化及其影响机制。方法使用含12000个与代谢、凋亡、细胞粘附、信号传导、转录因子等有关基因的cDNA表达谱芯片,检测4例重度妊娠高血压疾病子痫前期及正常的胎盘组织的基因表达谱差异,并差异表达的细胞代谢相关基因进行了Northern验证。结果4先兆子痫胎盘中同时有44种基因表达发生了变化,其中糖原磷酸化酶（GP—M）基因、瘦素（1eptin）基因、脂蛋白酯酶（LPL）基因及糖代谢相关基因（Glucose transporter）表达增强,核苷酸代谢基因（CD73）与能量代谢调节基因（Creatine kinase B）表达下降。结论多种基因表达异常与先兆子痫的病理发生有关,细胞代谢相关基因表达异常可能是血管内皮损伤的原因之一。     

川芎嗪对脂多糖诱导大鼠心肌细胞炎症及氧化应激的作用

背景:脓毒症心肌损伤与炎症反应及氧化应激损伤有关。既往研究表明川芎嗪具有抗氧化、维持钙稳态等多种功能。目的:探讨川芎嗪对脂多糖诱导的大鼠心肌细胞炎症反应及氧化应激作用的影响。方法:利用脂多糖(1μg/mL)诱导大鼠心肌细胞建立脓毒症心肌细胞炎性模型,通过不同浓度的川芎嗪(40,80,160μmol/L)干预治疗,24 h后进行检测。利用试剂盒检测川芎嗪对脂多糖诱导H9C2细胞乳酸脱氢酶以及活性氧释放的影响,荧光显微镜观察细胞凋亡情况,检测炎症因子白细胞介素1β、肿瘤坏死因子α和白细胞介素6及Toll样受体4、MyD88的表达变化。结果与结论:(1)与对照组比较,模型组细胞中乳酸脱氢酶、活性氧表达水平显著上升(P <0.05),荧光显微镜下观察发现大量细胞被PI染色,且白细胞介素1β、肿瘤坏死因子α和白细胞介素6的表达水平显著升高(P <0.05);Toll样受体4、MyD88基因表达均显著上升(P <0.05);(2)与模型组比较,川芎嗪治疗组能够有效降低乳酸脱氢酶释放,降低活性氧水平,抑制PI染色H9C2细胞数目,降低白细胞介素1β、肿瘤坏死因子α和白细胞介素6的表...     

不育症患者精浆IL-2,IL-6,IL-8,TNF-α测定的临床意义

目的 :观察精浆中白细胞介素 - 2 (IL - 2 )、白细胞介素 - 6 (IL - 6 )、白细胞介素 - 8(IL - 8)、肿瘤坏死因子 -α(TNF -α)等细胞因子对男性生殖的影响。方法 :应用放射免疫分析 (RIA)对 1 2 6例不育男性和 2 0例正常生育男性精浆IL - 2、IL - 6、IL - 8、TNF -α水平进行了检测。结果 :不育症组精浆IL - 2、IL - 6、IL -8、TNF -α含量均高于生育组 (p <0 0 5或 p <0 0 1 ) ;精浆中IL - 2、IL - 6、IL - 8、TNF -α含量在不育症组的WBC精液组与非WBC精液组之间均存在显著性差异 (p <0 0 5或 p <0 0 1 )。另外 ,精浆中TNF -α含量在精子活动力、活动率正常与减少之间 ,IL - 8含量在精子活动率正常与减少之间均存在显著性差异 (p <0 0 5或p <0 0 1 )。结论 :检测精浆中IL - 2、IL - 6、IL - 8、TNF -α等细胞因子的含量可以反映男性不育症患者的状态 ,帮助临床进行有价值的治疗     

Increased T-helper-1-type immunity and decreased T-helper-2-type immunity in patients with preeclampsia.

PROBLEM: To examine whether preeclampsia involves type-1 T-helper (TH1) immune hyperactivity. METHOD OF STUDY: Expression of HLA-DR, a cell-surface marker of activation, was analyzed on CD3+, CD4+, and CD8+ T cells in 15 preeclamptic patients and 15 normal pregnant women using flow cytometry. Additionally, peripheral blood mononuclear cells from preeclamptic patients and normal pregnant women were cultured with or without phytohemagglutinin (PHA) stimulation, and interleukin (IL)-2, IL-4, interferon (IFN)-gamma, and tumor necrosis factor (TNF)-alpha concentrations were determined in the supernatant by immunoassays. RESULTS: HLA-DR antigen was expressed more strongly on CD3+ T cells in preeclamptic patients than in normal subjects. In preeclampsia, HLA-DR was expressed more strongly in CD8+ T cells than in CD4- T cells. More TNF-alpha, IL-2, and IFN-gamma were produced by unstimulated and stimulated cultured peripheral blood mononuclear cells from preeclampsia patients than by those from normal subjects. TNF-alpha/IL-4, IL-2/IL-4, and IFN-gamma/IL-4 ratios were higher in preeclamptic patients than in the normal group. Significant positive correlations were observed between mean blood pressure and concentrations of the Th-1 type cytokines IL-2, IFN-gamma, and TNF-alpha. CONCLUSION: Up-regulation of Th1 responses and down-regulation of Th2 responses occur in preeclampsia.     

Spontaneous and induced release of prostaglandins, interleukin (IL)-1beta, IL-6, and tumor necrosis factor-alpha by placental tissue from normal and preeclamptic pregnancies

PROBLEM: The objective of this study was to clarify the role of the main proinflammatory cytokines (interleukin [IL]-1, IL-6, tumor necrosis factor [TNF]-alpha) in the pathogenesis of preeclampsia and how these cytokines affect one another and the production of prostaglandins (PGs). METHOD OF STUDY: The concentrations of cytokines and PGs in supernatants of placental tissue from preeclamptic and normal women were determined by enzyme-linked immunosorbent assay. RESULTS: The concentrations of the PGs from unstimulated preeclamptic placental tissue were significantly higher compared to the concentrations of PGs from normal unstimulated placental tissue. Significant levels of IL-1beta were observed only in the supernatants of preeclamptic placental tissue. Of interest, an increase in TNF-alpha production was detected in the supernatants of IL-1-stimulated preeclamptic placental tissue. CONCLUSIONS: The overproduction of TNF-alpha may be related not only to the effect of a stimulant like IL-1beta, but mainly to the lack of mechanisms down-regulating the production of TNF-alpha.     

Elevated IL-6 in Midtrimester Amniotic Fluid Is Involved with the Onset of Preeclampsia

PROBLEM: The primary defect of placental development in preeclampsia is speculated to occur at midtrimester gestation. Abnormal feto-maternal immune reactions have been considered as factors in such defective placentation. METHOD OF STUDY: Midtrimester amniotic fluid specimens were retrospectively identified as coming from gestations that later had severe preeclampsia develop, gestations with normal outcomes, and gestations measured for cytokines tumor necrosis factor-α (TNF-α), interleukin (IL-1β, IL-6, and IL-8). The effect of each cytokine on thrombomodulin levels was tested in cultured trophoblast cells. RESULTS: Among the measured cytokines, IL-6 and IL-8 were significantly elevated in the midtrimester amniotic fluid of the future preeclamptic group. Trophoblasts stimulated with TNF-α plus IL-6 had significantly decreased levels of cellular thrombomodulin compared to those without cytokine addition. CONCLUSIONS: Elevated cytokines in midtrimester amniotic fluid suggest an abnormal fetomaternal immune response occurring before the clinical manifestation of preeclampsia. Cytokine-induced suppression of thrombomodulin in trophoblasts may be directly involved in the pathogenesis of preeclampsia.     

Circulating Levels of Immunoreactive Cytokines in Women with Preeclampsia

PROBLEM: Circulating inflammatory cytokines have been implicated in the pathogenesis of preeclampsia. To test this hypothesis, we measured plasma levels of immunoreactive tumor necrosis factor (TNF)-α and -β, interleukin (IL)?1α and -β, and IL-6 and ?10 in women with preeclampsia, in women with transient gestational hypertension, and throughout normal pregnancy. METHOD OF STUDY: Enzyme-linked immunosorbent assays were used and subjected to extensive validation studies. RESULTS: The median concentration of plasma TNF-α was increased by twofold in women with preeclampsia compared with that in normal third-trimester pregnancy (P < 0.001) and in women with gestational hypertension (P < 0.04). The median concentration of plasma IL-6 was increased by threefold in women with preeclampsia compared with that in normal third-trimester pregnancy (P < 0.001) and increased twofold compared with that in women with gestational hypertension (P < 0.1). There were no significant differences observed in the levels of plasma IL-1β and IL-10 between the preeclamptic and other subject groups. The level of IL-1β, but not the levels of IL-10, TNF-α, or IL-6, was significantly changed during normal pregnancy compared with the nonpregnant condition manifesting an overall decline (P < 0.04). TNF-β and IL-1α were not detected in any samples, possibly because of the low sensitivity of these particular immunoassays. CONCLUSION: Elevated levels of TNF-α and IL-6 may contribute to the putative endothelial dysfunction of preeclampsia.     

Preeclampsia-related inflammatory cytokines regulate interleukin-6 expression in human decidual cells

Preeclampsia, a common pregnancy disorder associated with an increase in systemic inflammation, is the leading cause of maternal and fetal morbidity and mortality throughout the world. It is associated with shallow extravillous trophoblast invasion of the decidua, leading to uteroplacental blood flow that is inadequate for the developing fetal-placental unit. In preeclamptic women, interleukin-6 (IL-6) levels in plasma, but not placenta, are elevated, prompting evaluation of the decidua as a potential source of this excess, circulating IL-6. The current study found significantly higher immunohistochemical staining for IL-6 in decidual cells from preeclamptic versus preterm, gestational age-matched control placentas. Pro-inflammatory cytokines associated with the genesis of preeclampsia (i.e., tumor necrosis factor-alpha and interleukin-1beta) enhanced IL-6 mRNA levels and increased secreted IL-6 levels in first trimester leukocyte-free decidual cell incubations, as measured by real time quantitative RT-PCR, ELISA, and Western blotting. Therefore, decidual cell-derived IL-6 may contribute to excess circulating IL-6 levels that can promote both endothelial cell dysfunction (and subsequent vascular dysfunction) and the pathogenesis of preeclampsia whereas locally elevated IL-6 levels may contribute to an excess of decidual macrophages implicated in shallow extravillous trophoblast invasion of the decidua.     

Evidence for an Elevation in Serum Interleukin-2 and Tumor Necrosis Factor-α Levels Before the Clinical Manifestations of Preeclampsia

PROBLEM: The purpose of this study is to clarify whether the disruption of immune regulation occurs in early pregnancy before the clinical manifestations of preeclampsia. METHOD OF STUDY: The serum concentrations of interleukin-2 (IL-2) and tumor necrosis factor-α (TNF-α) were determined by using enzyme-linked immunoadsorbent assay (ELISA) in the first trimester of pregnancy in women who had preeclampsia develop after 28 weeks of pregnancy (preeclamptic group) and in women who completed pregnancy uneventfully (control group). RESULTS: Serum concentrations of both IL-2 and TNF-α in the first trimester of the preeclamptic group were significantly higher than those of the control group. CONCLUSIONS: That the perturbation of feto-maternal immune regulation may precede the clinical manifestations of preeclampsia, which may be of relevance in the development of preeclampsia, is suggested.     

Neutrophil NETs: a novel contributor to preeclampsia-associated placental hypoxia?

Recent studies have suggested that the innate immune system is involved in the pathogenesis of preeclampsia. Its pathogenesis involves neutrophil activation and increased levels of cell-free DNA in the maternal plasma. Activation of neutrophils has recently been shown to induce DNA containing neutrophil extracellular traps (NETs) which trap and kill bacteria. Massive NETs induction by the placentally derived factors (IL-8 and placental micro-debris) and their increased presence in preeclamptic placenta suggest that NETs might be involved in the pathogenesis of preeclampsia. Therefore, increased presence of NETs in preeclampsia may play a role in the deficient placental perfusion associated with this disorder.