Cyclin B1和p21在妊娠期高血压产妇胎盘组织中的表达及其临床意义

目的:观察CyclinB1和p21在妊娠期高血压产妇胎盘组织中的表达并探讨其临床意义.方法:采用免疫组化MaxVision法检测正常胎盘组织(20例)、妊娠期高血压胎盘组织(30例)、轻度子痫前期胎盘组织(30例)和重度子痫前期(30例)产妇胎盘组织中的cyclin B1和p21蛋白表达,并分析其与妊娠期高血压病情严重程度的相关性.结果:妊娠期高血压、轻度子痈前期、重度子痈前期胎盘组织中cyclinB1蛋白的表达均显著低于正常胎盘组织,差异均有统计学意义(P＜0.05),妊娠期高血压、轻度子痈前期、重度子痈前期胎盘组织中p21蛋白表达均显著高于正常胎盘组织,差异有统计学意义(P＜0.05).妊娠期高血压患者病情的严重程度与其胎盘组织中cyclinB1的蛋白表达呈显著负相关(r=0.641,P=0.000);而与其胎盘组织中p21蛋白的表达呈显著正相关(r=0.635,P=0.000).结论:Cyclin B1蛋白的表达下调和p21蛋白的表达上调可能参与了妊娠期高血压的发生发展,且二者在胎盘组织中的表达水平与妊娠期高血压病情的严重程度显著相关.     

慢性低氧高二氧化碳肺动脉高压小鼠肺组织Rho激酶的表达变化

慢性低氧高二氧化碳性肺动脉高压严重威胁着国民身体健康,但其发病机制尚未完全阐明。该研究通过检测正常对照组和慢性低氧高二氧化碳组小鼠右心室肥厚指数(right ven-tricular hypertrophy index,RVHI)、管壁厚度占血管外径的百分比(vessel wall thickness/total vasculardiameter,WT%)和管壁面积占血管总面积的百分比(vessel wall area/total vascular area,WA%),RT-PCR检测肺组织中Rho激酶(ROCK1,ROCK2)基因的表达,Western blot检测肺组织中ROCK1、p-MYPT1(phospho-myosin phosphatase target subunit 1)蛋白的表达,免疫组织化学法观察ROCK1的定位表达,探讨了Rho激酶在慢性低氧高二氧化碳性肺动脉高压形成中的作用。结果发现,慢性低氧高二氧化碳组小鼠RVHI、WT%、WA%值均显著升高(P<0.01),ROCK1、ROCK2基因表达明显增加(ROCK1 P<0.01,ROCK2 P<0.05),ROCK1、p-MYPT1蛋白表达显著增加(P<0.01),ROCK1蛋白表达于肺动脉、肺泡和支气管。以上结果提示,慢性低氧高二氧化碳条件下,小鼠肺组织中Rho激酶表达升高,可能参与了肺动脉高压的形成。     

RhoC和Ki67在非小细胞肺癌中的表达及意义

目的探讨RhoC、Ki67在非小细胞肺癌中的表达状况及其与临床病理学参数间的关系。方法利用免疫组织化学(ElivisionTMplus法),检测肺癌组织芯片中151例非小细胞肺癌患者RhoC、Ki67的表达情况。结果肺癌组织的RhoC蛋白阳性表达率为59.60%(90/151),相应癌旁组织RhoC阳性表达率为32.7%(16/49),差别有显著意义(P<0.05)。RhoC在TNMIII/IV期的NSCLC组织中阳性表达率为68.9%(51/74),明显高于在Ⅰ/II期NSCLC组织中阳性表达率50.6%(39/77,P<0.05);RhoC在出现淋巴结转移的表达率为49.2%(32/65),显著高于没有淋巴结转移的阳性表达率67.4%(58/86,P<0.05);非小细胞肺癌中RhoC高表达同Ki-67高表达呈显著正相关关系(χ2=21.634,r=0.377,P<0.01)。结论RhoC的高表达与非小细胞肺癌的分期,淋巴结转移及增殖有关。     

草鱼ROCK1基因全长克隆、生物信息学分析及组织差异表达

ROCK1是ROCK(即Rho激酶)家族的成员之一,ROCK家族是小G蛋白Rho的下游效应因子,被Rho蛋白激活后ROCK1作用于下游效应分子,对其介导的信号通路进行调节,参与细胞增殖、迁移、形态改变等。ROCK1与组织纤维化、癌症、心血管等疾病的发生密切相关。本实验采用c DNA末端快速扩增法克隆草鱼ROCK1基因全长序列,预测并分析其编码的蛋白,采用实时荧光定量PCR法检测草鱼ROCK1基因的组织差异表达。实验首次克隆获得了草鱼ROCK1基因全序列,长4 604 bp,编码1 361个氨基酸;草鱼ROCK1氨基酸序列与金线鲃、斑马鱼、鲤鱼ROCK1的氨基酸序列同源性分别达95%、94%、94%,同源性较高;草鱼ROCK1蛋白分子量158 035.73,理论等电点5.69,为亲水性非分泌蛋白,无信号肽;其二级结构包括α-螺旋(α-helix)、β-重叠(β-strand)和环(loop)。组织差异表达显示草鱼ROCK1基因在各组织中均有表达,其中在脑和血液中表达量最高(p0.05),其次为鳃、心脏、肾脏、脾脏和前肠组织,肌肉和肝脏组织表达量较低,皮肤组织表达量最低,所得结果将为后续研究草鱼组织纤维化奠定基础。     

七叶皂苷钠对细菌性脑膜炎大鼠RhoA/ROCK通路及血脑屏障通透性的影响CSCD

目的探讨七叶皂苷钠(SA)对细菌性脑膜炎(BM)大鼠肉瘤同源基因A(RhoA)/Rho相关螺旋卷曲蛋白激酶(ROCK)通路及血脑屏障通透性的影响。方法将SD新生大鼠随机分为正常对照组、模型组、SA低剂量(1.75 mg/kg)组、SA中剂量(3.50 mg/kg)组、SA高剂量(7.00 mg/kg)组和阳性对照(青霉素,0.18 g/kg)组,每组20只。除正常对照组外,其余各组大鼠均经小脑延髓池注射B族溶血性链球菌建立BM模型,建模成功后经腹腔注射给予相应剂量药物,连续给药3 d,2次/d。末次给药12 h后,处死大鼠,采用酶联免疫吸附试剂盒(ELISA)检测脑脊液中炎症因子白细胞介素-6(IL-6)、肿瘤坏死因子(TNF-α)水平;血细胞分析仪检测脑脊液中白细胞(WBC)数量;伊文思蓝(EB)染色法检测大鼠血脑屏障通透性;干湿比重法计算各组大鼠脑组织含水量;免疫组化法检测脑皮质组织RhoA、ROCK阳性表达水平;蛋白免疫印迹法(Western blotting)检测脑皮质组织血脑屏障相关分子闭锁连接蛋白-1(ZO-1)、紧密连接蛋白-5(Claudin-5)、水通道蛋白4(AQP4)及RhoA/ROCK通路相关蛋白肌球蛋白轻链(MLC)、丝切蛋白1(Cofilin1)相对表达水平。结果与正常对照组相比,模型组大鼠神经行为评分及脑皮质组织中ZO-1蛋白、Claudin-5蛋白、AQP4蛋白表达降低(均P<0.05),脑脊液WBC数量、IL-6水平、TNF-α水平、脑含水量、脑组织EB含量、脑皮质组织RhoA与ROCK阳性表达、p-MLC/MLC、p-Cofilin1/Cofilin1蛋白表达升高(均P<0.05)。与模型组相比,SA低、中、高剂量组与阳性对照组大鼠神经行为评分及脑皮质组织中ZO-1蛋白、Claudin-5蛋白、AQP4蛋白表达升高(均P<0.05),脑脊液WBC数量、IL-6水平、TNF-α水平、脑含水量、脑组织EB含量、脑皮质组织RhoA与ROCK阳性表达及p-MLC/MLC蛋白、p-Cofilin1/Cofilin1蛋白表达降低(均P<0.05);SA中、高剂量组上述指标变化优于SA低剂量组(P<0.05),阳性对照组上述指标变化弱于SA低剂量组(P<0.05);SA中剂量组与高剂量组相比差异无统计学意义(P>0.05)。结论SA可改善BM大鼠脑水肿及血脑屏障通透性,且其改善血脑屏障损伤的作用可能与抑制RhoA/ROCK通路激活有关。     

NF-kB 与Slug 在非小细胞肺癌及其上皮间质转化中的作用

目的:研究核因子NF-kB与slug在非小细胞肺癌(NSCLC)中的表达情况、及二者与非小细胞肺癌上皮间质转化(EMT)的关系,为非小细胞肺癌的诊断治疗提供理论依据。方法:(1)采用免疫组化PV9000二步法测定50例NSCLC组织及20例相应正常肺组织中NF-kBP65、slug、E-cadherin及Vimentin蛋白表达情况。(2)采用RT-PCR测定其中25例NSCLC组织及10例相应正常肺组织中NF-kBP65、slug的mRNA表达情况。结果:NSCLC中NF-kBP65蛋白表达量高于癌旁正常肺组织(Z=-2.370,P<0.05),NF-kBP65mRNA表达量明显高于癌旁正常肺组织(t=4.967,P<0.01);Slug蛋白表达量明显高于癌旁正常肺组织(Z=-4.443,P<0.01),SlugmRNA表达量明显高于癌旁正常肺组织(t=6.483,P<0.01)。在NF-kBP65阳性癌组织中,E-cadherin蛋白表达下降(x2=5.024,P<0.05),Vimentin蛋白表达上升(x2=4.723,P<0.05);Slug阳性癌组织中,E-cadherin蛋白表达下调(x2=5.984,P<0.05),Vimentin表达上调(x2=5.028,P<0.05)。另外,NF-kBP65与Slug在蛋白水平呈极显著正相关(r=0.443,P<0.01),在mRNA水平呈显著正相关(r=0.439,P<0.05)。NF-kB与分化程度(x2=5.024,P<0.05)、有无淋巴结转移(x2=7.933,P<0.01)及肿瘤的分期(x2=5.614,P<0.05)有关,与性别、年龄、组织类型无明显相关性(P>0.05);Slug与淋巴结转移(x2=6.174,P<0.05)及肿瘤的分期(x2=7.317,P<0.01)有关,与性别、年龄、组织类型、分化程度无明显相关性(P>0.05)。结论:NF-kB、Slug在NSCLC中表达增强,可能与NSCLC的发生、发展、转移有关;并且NF-kB与Slug可能协同抑制E-cadherin表达,促进Vimentin表达,诱使NSCLC的EMT发生,从而为进一步研究NSCLC的EMT提供理论依据。     

乳腺癌相关透明质酸变化对淋巴内皮细胞紧密连接分子ZO-1分布及细胞增殖的影响

该研究旨在探讨乳腺癌来源的透明质酸(hyaluronic acid,HA)分子大小变化对淋巴内皮细胞紧密连接分子ZO-1(zonula occludens-1)分布情况以及细胞增殖的影响。采用免疫组化实验检测8对良性乳腺疾病和乳腺癌患者组织HA的表达水平,利用酶联免疫吸附实验检测20对正常人与乳腺癌患者血清中透明质酸分解酶(hyaluronidase,Hyase)含量;通过免疫荧光法、荧光素钠渗透实验、MTT增殖实验和Western blot分别观察HA和寡分子透明质酸(oligosaccharides of Hyaluronic acid,o HA)作用淋巴内皮细胞后ZO-1分布、单层细胞渗透性、细胞增殖以及下游ROCK1/Rho A信号通路变化。结果表明,与良性乳腺疾病对照组比较,乳腺癌患者HA表达量明显增加(P0.01);与正常人相比,乳腺癌患者血清Hyase水平显著升高(P0.01),提示乳腺癌发生时,HA升高伴随其降解酶增多,HA代谢活性增加。HA增强细胞间ZO-1的连接,对ROCK1/Rho A蛋白表达水平无明显影响;相反,o HA促使淋巴内皮细胞ZO-1由胞膜向胞浆分布,导致细胞连接呈褶皱状,形成细胞间隙,淋巴管渗透性增加,上调ROCK1/Rho A蛋白表达水平,促进淋巴内皮细胞增殖(P0.01)。研究提示,o HA可能在乳腺癌淋巴管形态及增殖中发挥重要作用。     

血管内皮生长因子D及其受体flt-4和NO在妊娠高血压症中的表达及意义

目的:观察妊娠高血压综合征患者血液血管内皮生长因子(VEGF)、fms-样酪氨酸激-酶4fit-4)和胎盘一氧化氮(N0)的表达及其临床意义.方法:选择妊高征患者35例为妊高组;选择同期正常晚期妊娠妇女36例为对照组.采用酶联免疫吸附法测定各组孕妇血清VEGF及其受体fit-4的含量,采用硝酸还原法测定胎盘组织NO浓度.结果:妊高组患者血清VEGF及其受体flt-4水平、胎盘组织NO含量与对照组相比,明显降低(P<0.05);妊高组患者血清VEGF水平与胎盘组织NO含量呈正相关(r=0.723,P<0.05),flt-4的水平与胎盘组织NO含量呈正相关(r=0.61,P<0.05).结论:血清VEGF及其受体fit-4在妊娠高血压综合征患者发病中有重要作用,它们可能通过影响NO的合成进一步在妊高征发病中发挥作用.     

胃癌中RhoC和MMP-9的表达及其临床意义

目的探讨RhoC基因和基质金属蛋白酶9(matrixmetalloproteinases-9,MMP-9)基因在胃癌中的表达及其与临床病理因素之间的关系。方法采用免疫组化SP法检测120例胃癌组织中RhoC和MMP-9的表达。结果120例胃癌组织中RhoC和MMP-9阳性率分别为80.00%(96/120)、85.83%(103/120),两者表达存在相关性(P<0.05)。RhoC和MMP-9表达与胃癌的浸润深度、淋巴结转移和肿瘤临床分期有明显差异(P<0.05)。结论RhoC和MMP-9蛋白在胃癌中过量表达与胃癌的发生发展密切相关,检测RhoC和MMP-2蛋白表达对于评估胃癌的预后有一定意义。     

ROCK Ⅰ/Ⅱ基因下调对血管平滑肌细胞迁移及增殖的影响

目的:探讨ROCK亚型ROCK Ⅰ和ROCKⅡ对血管平滑肌细胞(A7r5)迁移及增殖的影响.方法:利用Western blot技术检测ROCK Ⅰ和ROCKⅡ蛋白在A7r5细胞中的表达水平;利用siRNA技术使ROCK Ⅰ和ROCKⅡ基因表达分别下调,并检测基因下调后蛋白表达水平;利用Boyden小室法,观察ROCK Ⅰ和ROCKⅡ基因下调后及RocK特异抑制剂Y-27632对PDGF诱导的A7r5细胞迁移的影响:使用MTT法检测ROCK Ⅰ和ROCKⅡ基因下调后对A7r5细胞生长曲线的影响.结果:ROCK Ⅰ和ROCKⅡ在A7r5细胞中的蛋白表达水平不同,ROCKⅡ较ROCK Ⅰ的表达水平高4倍;通过对A7r5细胞进行ROCK Ⅰ和ROCKⅡ siRNA转染,使二者蛋白表达水平分别下调83.4%和94.7%;基因表达下调后,ROCK Ⅰ明显抑制了PDGF诱导的A7r5细胞的迁移,而ROCKⅡ无明显影响,Y-27632也抑制了A7r5细胞的迁移;ROCK Ⅰ和ROCKⅡ基因下调后对A7r5细胞生长曲线的影响无明显差别.结论:ROCK Ⅰ在血管平滑肌细胞迁移过程中起主导作用,ROCK Ⅰ和ROCKⅡ对血管平滑肌细胞的增殖作用无明显差异.     

Ser1333 phosphorylation indicates ROCKI activation

Background

Two isoforms of Rho-associated protein kinase (ROCK), ROCKI and ROCKII, play a pivotal role in regulation of cytoskeleton and are involved in multiple cellular processes in mammalian cells. Knockout mice experiments have indicated that the functions of ROCKI and II are probably non-redundant in physiology. However, it is difficult to differentiate the activation status of ROCKI and ROCKII in biological samples. Previously, we have identified phosphorylation site of ROCKII at Ser1366 residue sensitive to ROCK inhibition. We further investigated the activity-dependent phosphorylation site in ROCKI to establish the reagents that can be used to detect their individual activation.

Results

The phosphorylation site of ROCKI sensitive to its inhibition was identified to be the Ser1333 residue. The ROCKI pSer1333-specific antibody does not cross-react with phosphorylated ROCKII. The extent of S1333 phosphorylation of ROCKI correlates with myosin II light chain phosphorylation in cells in response to RhoA stimulation.

Conclusions

Active ROCKI is phosphorylated at Ser1333 site. Antibodies that recognize phospho-Ser1333 of ROCKI and phospho-S1366 residues of ROCKII offer a means to discriminate their individual active status in cells and tissues.     

Determination of potential selective inhibitors for ROCKI and ROCKII isoforms with molecular modeling techniques: structure based docking,ADMET and molecular dynamics simulation

Rho-associated protein kinases (ROCKs) are a member of the serine/threonine protein kinase family and potential therapeutic target for various diseases. This enzyme has two isoforms, Rho-associated protein kinase I (ROCKI) and Rho-associated protein kinase II (ROCKII). They share an overall 65% homology in all amino acid sequence and 92% homology in kinase domains. Since, the kinase domains of ROCKI and ROCKII are highly conserved and similar, the discovery and design of isoform-selective inhibitors are more challenging. Thus, most currently available agents that is against ROCKs exhibit low selectivity and severe side effects. Therefore, this study aimed to elucidate the interaction of compounds that indicated high potential in experimental studies against ROCKI and ROCKII enzymes in the molecular level with molecular modeling techniques. Firstly, we determined the interaction property of catalytic sites of the ROCKs by analyzing with molecular docking. Based on these results, the best ligands (50 compounds) corresponding to experimental studies were selected, and then absorption, distribution, metabolism and excretion – toxicity (ADMET) analysis of these compounds were implemented. According to these study results, the compound 40 for ROCKI and the compound 50 for ROCKII were identified as selective and highly potent inhibitors. And finally, molecular dynamics (MD) simulations were performed for the stability of ROCKs with identified compounds. In the light of this study, it will be possible to treat diseases that ROCKs have a role by developing more effective and specific ROCK inhibitors.

Communicated by Ramaswamy H. Sarma     

miR-143 decreases prostate cancer cells proliferation and migration and enhances their sensitivity to docetaxel through suppression of KRAS

As there is increasing evidence that Rho-Rho kinase (ROCK) pathway plays an important role in the proliferation and contraction in many tissues, we investigated the contractile role of a ROCK inhibitor, fasudil, and the distribution of RhoA, RhoB, RhoC, ROCK1, and ROCK2 in the rat prostate. Twelve-week-old Sprague-Dawley rat prostate was used in this study. Rat prostatic contractile responses induced by carbachol and norepinephrine were investigated in organ bath studies without or with 10(-7), 10(-6), and 10(-5) M of a non-selective ROCK inhibitor, fasudil. Immunoblot analysis and immunohistochemical staining were performed to investigate the participation levels of RhoA, RhoB, RhoC, ROCK1, and ROCK2. The E(max) values induced by carbachol and norepinephrine were similar in the rat prostate. Fasudil significantly inhibited carbachol- or norepinephrine-induced prostatic contractions in a dose-dependent manner. Fasudil 10(-5) M reduced the initial prostatic contraction (without fasudil) to 56.7 ± 5.9% for carbachol and to 45.7 ± 12.3% for norepinephrine. Amounts of RhoA, RhoB, RhoC, ROCK1, and ROCK2 were detected by immunoblot analysis in the prostate. Immunohistochemical study revealed that RhoA, RhoB, RhoC, ROCK1, and ROCK2 were all positive in the prostatic smooth muscle, while there were some differences of distributions of Immunoreactivities between these enzymes in the prostatic glandula. Our data indicated that rat prostate contains RhoA, RhoB, RhoC, ROCK1, and ROCK2, which play an important role in the autonomic nerve-mediated contractile responses in the prostate.     

血清血管生成素相关生长因子水平与子痫前期发病的关系

目的:探讨血清血管生成素相关生长因子(AGF)水平与子痫前期发病的关系。方法:选择2013年1月至2014年12月间在我院进行产前检查并行剖宫产终止妊娠的子痫前期患者42例作为研究组,同期收住的正常妊娠孕产妇30例作为对照组。应用双抗体酶联免疫吸附法检测血清AGF水平,应用RT-PCR法测定胎盘组织AGF mRNA水平。结果:研究组收缩压、舒张压及血清AGF水平显著高于对照组,分娩孕周显著小于对照组,新生儿出生体重显著低于对照组,差异有统计学意义(P0.05)。研究组胎盘AGF mRNA水平显著高于对照组,差异有统计学意义(P0.05)。多因素Logistic分析显示,孕产妇胎盘AGF mRNA水平与血清AGF水平、孕产妇收缩压、舒张压呈正相关(r=0.605,0.428,0.403,P均0.05)。结论:子痫前期患者血清AGF水平异常,胎盘AGF表达异常,提示AGF可能与子痫前期发病有关。     

ROCK I-mediated activation of NF-kappaB by RhoB

RhoB is a short-lived protein whose expression is increased by a variety of extra-cellular stimuli including UV irradiation, epidermal growth factor (EGF) and transforming growth factor beta (TGF-beta). Whereas most Rho proteins are modified by the covalent attachment of a geranylgeranyl group, RhoB is unique in that it can exist in either a geranylgeranylated (RhoB-GG) or a farnesylated (RhoB-F) form. Although each form is proposed to have different cellular functions, the signaling events that underlie these differences are poorly understood. Here we show that RhoB can activate NF-kappaB signaling in multiple cell types. Whereas RhoB-F is a potent activator of NF-kappaB, much weaker activation is observed for RhoB-GG, RhoA, and RhoC. NF-kappaB activation by RhoB is not associated with increased nuclear translocation of RelA/p65, but rather, by modification of the RelA/p65 transactivation domain. Activation of NF-kappaB by RhoB is dependent upon ROCK I but not PRK I. Thus, ROCK I cooperates with RhoB to activate NF-kappaB, and suppression of ROCK I activity by genetic or pharmacological inhibitors blocks NF-kappaB activation. Suppression of RhoB activity by dominant-inhibitory mutants, or siRNA, blocks NF-kappaB activation by Bcr, and TSG101, but not by TNFalpha or oncogenic Ras. Collectively, these observations suggest the existence of an endosome-associated pathway for NF-kappaB activation that is preferentially regulated by the farnesylated form of RhoB.     

RhoC/ROCK2 promotes vasculogenic mimicry formation primarily through ERK/MMPs in hepatocellular carcinoma

Vasculogenic mimicry (VM) results in the formation of an alternative circulatory system that can improve the blood supply to multiple malignant tumors, including hepatocellular carcinoma (HCC). However, the potential mechanisms of RhoC/ROCK in VM have not yet been investigated in HCC. Here, RhoC expression was upregulated in HCC tissues, especially the VM-positive (VM+) group, compared to noncancerous tissues (P < 0.01), and patients with high expression of RhoC had shorter survival times (P < 0.001). The knockdown of RhoC via short hairpin RNA (shRNA) in SK-Hep-1 cells significantly decreased VM formation and cell motility. In contrast, cell motility and VM formation were remarkably enhanced when RhoC was overexpressed in HepG2 cells. To further assess the potential role of ROCK1 and ROCK2 on VM, we stably knocked down ROCK1 or ROCK2 in MHCC97H cells. Compared to ROCK1 shRNA, ROCK2 shRNA could largely affect VM formation, cell motility and the key VM factors, as well as the epithelial-mesenchymal transition (EMT) markers in vitro and in vivo. Moreover, p-ERK, p-MEK, p-FAK, p-paxillin, MT1-MMP and MMP2 levels were clearly altered following the overexpression of RhoC, but ROCK2 shRNA had little effect on the expression of p-FAK, which indicated that RhoC regulates FAK/paxillin signaling, but not through ROCK2. In conclusion, our results show that RhoC/ROCK2 may have a major effect on VM in HCC via ERK/MMPs signaling and might be a potential therapeutic target for the treatment of HCC.     

IL-4 和IFN-γ mRNA 在子痫前期患者胎盘组织中的表达及意义

目的:检测胎盘组织中IFN-γ和IL-4的表达,探讨IFN-γ和IL-4在子痫前期的发病中的作用.方法:采用原位杂交法检测了20例正常妊娠孕妇和34例子痫前期组(包括16例轻度和18例重度)中的IFN-γ、IL-4 mRNA的表达水平,并通过图像分析系统对染色结果进行定量分析.结果:(1)IL-4 mRNA的表达在正常妊娠组、子痫前期轻度组和子痫前期重度组的表达无差异(P＞0.05).(2)与正常妊娠组、子痫前期轻度组相比,子痫前期重度组IFN-γ mRNA的表达有差异性(P＜0.05);子痫前期轻度组与正常妊娠组相比无差异(P＞0.05).(3)与正常妊娠组相比,子痫前期轻度组、重度组IFN-γ mRNA/IL-4 mRNA的比值均有差异性(P＜0.05),且随病情的加重比值增大.结论:Th1/Th2细胞因子的平衡偏离可能是导致子痫前期发病的病因之一.     

miR-19a 在正常妊娠及重度子痫前期患者胎盘中的表达

目的:通过检测正常妊娠及重度子痫前期患者胎盘组织中miR-19a的表达,探讨其与子痫前期发病的关系。方法:收集10例重度子痫前期患者胎盘组织(实验组)和10例正常产妇胎盘组织(对照组),应用荧光实时定量PCR(Real Time PCR)的方法检测两组miR-19a的表达差异。结果:重度子痫前期患者胎盘组织中miR-19a表达升高(P<0.05)。结论:子痫前期患者胎盘组织中存在差异表达的miRNA,miR-19a在胎盘组织中的高表达可能与子痫前期的发病有关。     

PEDF蛋白在子痫前期患者胎盘上的表达及意义

目的:研究子痫前期患者胎盘组织中色素上皮衍生因子(pigment epithelium-derived factor,PEDF)的表达,探讨PEDF在子痫前期发病中的作用。方法:选取2012年3月至2013年3月在我院产科住院剖宫产的20例子痫前期孕妇作为研究对象,另选取同期正常分娩的孕妇20例作为对照组。采用Western blot、免疫荧光组织化学方法检测子痫前期患者和正常对照组妇女胎盘组织中PEDF和血管内皮生长因子(vascular endothelial growth factor,VEGF)的表达,并通过免疫荧光方法计数胎盘微血管密度(MVD)。结果:同正常对照组相比,子痫前期患者胎盘组织中PEDF表达升高,而VEGF表达则减少。PEDF与VEGF组织表达位置大致相同。子痫前期患者胎盘组织中PEDF表达与24h尿蛋白定量呈正相关,与VEGF表达及MVD计数呈负相关;VEGF表达与MVD计数呈正相关。结论:PEDF与子痫前期疾病发生发展及病情轻重程度有关;PEDF可能是通过影响胎盘血管的重铸,而参与子痫前期疾病的发生发展。