沙利度胺对实验性矽肺血管生成的干预研究

目的观察实验性矽肺形成过程中肺组织局部血管生成的动态变化,以及沙利度胺对矽肺肺纤维化血管生成的干预作用。方法SD大鼠54只,随机分为3组,矽肺组和沙利度胺组气管内注入二氧化硅混旋液复制实验性大鼠矽肺模型,对照组在相同条件下给予生理盐水。第二天起沙利度胺组给予沙利度胺饲料喂养,其余各组在相同条件下给予普通饲料喂养。采用HE染色、羟脯氨酸含量测定、免疫组织化学染色等方法,观察实验性大鼠矽肺的发病过程,肺组织中p-Akt蛋白和局部血管生成的动态变化及其与肺胶原蛋白含量的关系。结果矽肺组第7天新生血管明显增多,第30天较第7天有所减少,到第60天肺组织正常结构基本消失,取代为广泛结节性纤维化,少见血管;沙利度胺组血管生成在第7天轻于矽肺组,但仍高于对照组。免疫组化显示p-Akt蛋白在矽肺组第7天明显达到高峰,第30天时较第7天有所减弱,第60天时明显减弱。沙利度胺组p-Akt蛋白和羟脯氨酸含量均低于矽肺组,但高于对照组。结论矽肺早期血管生成显著,血管生成在矽肺肺纤维化发生早期可能起重要作用;沙利度胺能在一定程度上抑制过度的血管生成,对实验性矽肺具有一定的阻抑作用。     

沙利度胺配合化疗应用于急性白血病患者的疗效评价

目的:探究急性白血病患者给予沙利度胺配合化疗在抗血管生长方面的临床成效。方法:选取我院2009年3月-2014年1月收治的86例急性白血病患者,随机分为研究组和对照组,每组43例。对照组患者给予常规化疗方案,研究组在对照组基础上给予沙利度胺配合化疗。观察两组患者治疗前后血浆VEGF,VEGFR,b FGF及MVD的水平变化。比较两组患者的临床疗效及不良反应发生率。结果:治疗前,两组患者VEGF、VEGFR、b FGF及MVD水平无显著差异(P0.05);治疗后,研究组患者VEGF、VEGFR、b FGF及MVD水平均低于对照组,差异有统计学意义(P0.05)。研究组患者治疗的有效率为88.4%,对照组为76.7%,研究组显著优于对照组,差异具有统计学意义(P0.05)。研究组不良反应发生率为79.1%,对照组为81.4%,差异无统计学意义(P0.05)。结论:沙利度胺配合化疗治疗急性白血病能调控促血管生长因子水平,提高疗效,不良反应可耐受。     

白芨中萜类化合物通过诱导血管内皮细胞凋亡抑制血管生成

本文研究了白芨中的萜类化合物对血管生成的抑制作用．及其抑制血管生成的可能机制。采用萃取和色谱法从白芨中分离和纯化了该萜类化合物。通过鸡胚绒毛囊膜（CAM）和人脐静脉内皮细胞（HUVEC）研究了白芨中萜类化合物及其粗提物对血管及血管内皮细胞的抑制作用。结果表明,含该萜类的粗提物显著抑制鸡胚绒毛尿囊膜血管生成;该萜类纯品能明显抑制HUVEC增殖,且可诱导HUVEC凋亡,包括细胞体积缩小,细胞膜起泡,细胞核裂解,染色质浓缩和边集,出现凋亡小体,DNA降解。因此．白芨萜类化合物的抗血管生成作用与诱导血管内皮细胞凋亡有关。     

沙利度胺治疗中晚期非小细胞肺癌的临床观察

目的:观察沙利度胺治疗中晚期非小细胞肺癌患者的预后情况.方法:沙利度胺治疗组选取30例中晚期非小细胞肺癌患者入组研究,给予沙利度胺联合PCDE方案治疗;同时选取37例中晚期非小细胞肺癌患者作为对照组,仅使用PCDE方案治疗,分析两组患者生存率的差异.结果:沙利度胺联合PCDE方案治疗组非小细胞肺癌患者的生存率明显高于PCDE方案治疗组患者(P<0.01).结论:采用沙利度胺联合PCDE方案治疗中晚期非小细胞肺癌,可以提高非小细胞肺癌患者的生存率,值得临床推广.     

鼠源休眠蛋白具有抑制内皮细胞增殖和诱导细胞凋亡的活性

内皮抑素 (endostatin)是最近发现的一种具有抑制血管生长的蛋白质。休眠蛋白 (restin)是内皮抑素的同源蛋白质 ,最早由Ramchandran等人发现 ,它来源于胶原蛋白XV的羧端非胶原蛋白结构域 (NC1)。为了研究鼠源休眠蛋白对内皮细胞生长的影响 ,利用RT PCR从鼠肌肉中扩增出它的基因 ,克隆入原核表达载体pQE32。诱导后 ,重组蛋白质以包涵体形式高效表达 ,表达量约占菌体总蛋白质的 6 0 %～ 70 %。重组蛋白质经纯化复性后 ,可以特异地抑制bFGF刺激的牛主动脉内皮细胞的增殖 ,但是休眠蛋白的抑制活性比内皮抑素活性稍低。流式细胞仪检测发现 ,休眠蛋白可以引起内皮细胞的细胞周期的改变 ,并且引起细胞凋亡     

沙利度胺抑制肿瘤细胞分泌VEGF/bFGF机制的探讨*

目的: 探讨Cereblon(CRBN)对沙利度胺抑制人肺癌A549细胞及人肝癌HepG2细胞分泌VEGF/bFGF的影响。方法: 采用慢病毒介导的短发夹RNA(shRNA)干扰技术建立稳定敲低CRBN的A549细胞系(A549_CRBN)及HepG2细胞系(HepG2_CRBN)并通过实时定量PCR(Real-time PCR)和蛋白质印记(Western blot)实验验证。将A549细胞分为阴性对照组(A549_luciferase)、CRBN低表达组(A549_CRBN);HepG2细胞分为阴性对照组(HepG2_luciferase)、CRBN低表达组(HepG2_CRBN),以上细胞按照 3×10⁵ cells/well接种到6孔板中,放入37℃,5%CO2的培养箱中培养24 h,分别加入1 ml含100 μmol/L沙利度胺(thalidomide组)和1 ml 1‰ DMSO(control组)的培养液,继续培养24 h再行后续实验,每组设计3个复孔。MTS法检测沙利度胺对细胞增殖的影响;Real-time PCR检测VEGF、bFGF、c-jun mRNA表达,ELISA法检测VEGF、bFGF蛋白表达。结果: 与对照组比较,沙利度胺在浓度为1、10、50、100 μmol/L 时对A549 及HepG2细胞的增殖能力无显著影响(P＞0.05)。与A549_CRBN或HepG2_CRBN组比较,A549_luciferase及HepG2_luciferase组分泌的VEGF及bFGF均显著降低(P＜0.05)。与A549_luciferase或HepG2_luciferase细胞的对照组比较,沙利度胺可抑制A549_luciferase和HepG2_luciferase细胞的VEGF和bFGF的表达(P＜0.05),而对A549_CRBN和HepG2_CRBN细胞中VEGF和bFGF的表达无显著抑制作用;与HepG2_luciferase细胞的对照组比较,沙利度胺可抑制HepG2_luciferase细胞的c-Jun表达(P＜0.01),而对HepG2_CRBN细胞的c-Jun表达无显著抑制作用。结论: 沙利度胺对A549和HepG2细胞VEGF和bFGF表达的抑制作用可能是通过CRBN介导的,而c-Jun可能是抑制作用的关键转录因子之一。     

出血性蛇毒诱导血管内皮细胞凋亡

蛇毒是自然界中活性蛋白质含量最为丰富的生物资源之一。出血性蛇毒含有丰富的作用于血液和血管系统的活性物质,其中包括多种具有诱导内皮细胞凋亡的组分,如金属蛋白酶和L-氨基酸氧化酶。该文主要介绍诱导内皮细胞凋亡的蛇毒组分,以及与之相关的血管内皮细胞凋亡的机制。对调节内皮细胞生存和凋亡的分子机制的了解,将有助于开发新的治疗肿瘤的药物或者方法。     

索拉非尼和沙利度胺对肝癌患者血清中VEGF-C、VEGF-D及微血管密度的影响

目的:探讨索拉非尼和沙利度胺这两种不同的化疗药物,对肝癌患者血清中VEGF-C、VEGF-D及微血管密度的影响。方法:将患者分成3组,每组16例。对照组采用常规治疗并服用安慰剂;索拉非尼和沙利度胺这两个组患者中,前者服用索拉非尼400 mg/次,2次/d,治疗6个月;后者服用沙利度胺每日服200mg,每周增加200mg/d,直至最大剂量每日600mg,至少服用4月。ELISA检测患者血清中VEGF-C、VEGF-D;免疫组织化学检测肝癌组织中微血管密度。结果:对照组患者血清中VEGF-C的水平为210ng/ml,索拉非尼组患者血清中VEGF-C的水平为132ng/ml,而沙利度胺组患者血清中VEGF-C的水平为186ng/ml。与对照组相比,索拉非尼组和沙利度胺组患者血清中VEGF-C的水平均降低。对照组患者血清中VEGF-D的水平为322ng/m1,索拉非尼组患者血清中VEGF-D的水平为217ng/ml,而沙利度胺组患者血清中VEGF-D的水平为256ng/ml。与对照组相比,索拉非尼组和沙利度胺组患者血清中VEGF-D的水平均降低。索拉非尼组患者血清中VEGF-D的水平明显低于沙利度胺高(P<0.05)。对照组肝癌组织MVD为(44.32±5.16)个,索拉非尼组患者肝癌组织MVD为(21.75±1.49)个,而沙利度胺组患者肝癌组织MVD为(34.78±2.31)个。结论:多靶点化疗药物索拉非尼对肝癌患者血清中VEGF-C、VEGF-D及微血管密度的影响最大,深入探讨其作用机制,可为其肝癌患者提供新的化疗方案。     

索拉非尼和沙利度胺对肝癌患者血清中VEGF-C、VEGF-D及微血管密度的影响

目的：探讨索拉非尼和沙利度胺这两种不同的化疗药物,对肝癌患者血清中VEGF-C、VEGF—D及微血管密度的影响。方法：将患者分成3组,每纽16例。对照组采用常规治疗并服用安慰剂;索拉非尼和沙利度胺这两个组患者中,前者服用索拉非尼400mg／次,2次／d,治疗6个月;后者服用沙利度胺每日服200mg,每周增加200mg／d,直至最大剂量每日600mg,至少服用4月。ELISA检测患者血清中VEGF-C、VEGF-D;免疫组织化学检测肝癌组织中微血管密度。结果：对照组患者血清中VEGF-C的水平为210ng／ml,索拉非尼组患者血清中VEGF—C的水平为132ng／ml,而沙利度胺组患者血清中VEGF—C的水平为186ng／ml。与对照组相比,索拉非尼组和沙利度胺组患者血清中VEGF—C的水平均降低。对照组患者血清中VEGF—D的水平为322ng／ml,索拉非尼组患者血清中VEGF—D的水平为217ng／ml,而沙利度胺组患者血清中VEGF—D的水平为256ng／ml。与对照组相比,索拉非尼组和沙利度胺组患者血清中VEGF—D的水平均降低。索拉非尼组患者血清中VEGF—D的水平明显低于沙利度胺高（P〈0．05）。对照组肝癌组织MVD为（44．32±5．16）个,索拉非尼组患者肝癌组织MVD为（21．75±1．49）个,而沙利度胺组患者肝癌组织MVD为（34．78±2．31）个。结论：多靶点化疗药物索拉非尼对肝癌患者血清中VEGF—C、VEGF—D及微血管密度的影响最大,深入探讨其作用机制．可为其肝癌患者提供新的化疗方案。     

沙利度胺对原发性肝癌介入后血管内皮生长因子及生存质量的影响

目的:探讨沙利度胺对原发性肝癌(HCC)介入后血管内皮生长因子(VEGF)及生存质量的影响。方法:将2014年1月至2016年1月期间我院收治的拟行肝动脉栓塞化疗(TACE)的HCC患者60例按照随机数字表法分为A、B两组,各30例。A组患者采用TACE+沙利度胺治疗,每晚200 mg,口服一个月;B组患者单行TACE治疗。分别于治疗前、治疗后3周检测两组患者的VEGF水平,于治疗前、治疗后1个月应用生存质量评分量表(QOL)评价患者生存质量,并比较两组患者治疗前后的体重及治疗过程中的不良反应。结果:治疗前两组患者血清VEGF水平差异无统计学意义(P0.05)。治疗后两组患者VEGF水平均较治疗前升高,其中A组患者治疗后VEGF水平显著低于B组患者(P0.01)。治疗前两组患者QOL分数和体重比较差异无统计学意义(P0.05)。而治疗后两组患者的QOL分数较治疗前均有所降低,但A组患者的QOL分数显著高于B组患者(P0.05),而两组患者治疗后的体重无统计学差异(P0.05)。A组30例患者中有1例(3.33%)患者因严重嗜睡、2例(6.67%)患者因严重口腔干燥而终止口服沙利度胺,其余27例患者均能耐受。结论:沙利度胺联合TACE方法治疗HCC可降低VEGF水平和改善癌症患者的生存质量,值得在临床上推广使用。     

Propolis Suppresses Tumor Angiogenesis by Inducing Apoptosis in Tube-Forming Endothelial Cells

We have reported that propolis suppresses tumor-induced angiogenesis in vivo and in vitro, but antiangiogenic mechanism of propolis at cellular level remains unclear. In this study, we observed that propolis not only inhibited tube formation but also induced apoptosis of endothelial cells. These results suggest that propolis exerts its antiangiogenic effects at least in part through induction of apoptosis.     

Vascular Permeability Factor/Vascular Endothelial Growth Factor Inhibits Anchorage-Disruption-Induced Apoptosis in Microvessel Endothelial Cells by Inducing Scaffold Formation

Survival and proliferation of endothelial cells requires both growth factors and an appropriate extracellular matrix to which cells can attach. In the absence of either, endothelial cells rapidly undergo apoptosis. Thus, when human microvascular endothelial cells (HDMEC) are plated on a hydrophobic surface such as untreated polystyrene, they rapidly undergo apoptosis and die. The present study demonstrates that vascular permeability factor/vascular endothelial growth factor (VPF/VEGF), an endothelial cell-selective cytokine, inhibits apoptosis of HDMEC cultured on untreated polystyrene and induces these cells to adhere, spread, and proliferate. VPF/VEGF-induced HDMEC adhesion was time-dependent, requiredde novoprotein synthesis, and was inhibited by a soluble RGD peptide but not by an inhibitor of collagen synthesis. Under the conditions of these experiments, VPF/VEGF downregulated expression of collagen IV and fibronectin but did not change collagen I mRNA levels. VPF/VEGF-induced HDMEC adhesion was inhibited by antibodies to αvβ5 and vitronectin but not by antibodies to αvβ3. Other endothelial growth factors and cytokines such as bFGF, HGF, and TGFβ did not reproduce the VPF/VEGF effect. We suggest that VPF/VEGF induces endothelial cells to deposit a scaffolding (likely involving vitronectin) that allows them to attach to and proliferate on an otherwise nonsupportive surface (hydrophobic polystyrene) and in this manner serves as both a survival factor and a growth factor.     

TAK1 Prevents Endothelial Apoptosis and Maintains Vascular Integrity

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沉默ABCE1对神经胶质瘤细胞U251增殖、迁移和凋亡的影响

ABCE1是ATP结合盒蛋白亚家族成员之一,在病毒感染,细胞增殖,抗凋亡,翻译起始和核糖体生物发生等过程中有重要的作用。为了探讨ABCE1对神经胶质瘤细胞U251增殖、迁移和凋亡的作用,本研究通过实时荧光定量PCR和免疫印迹实验检测ABCE1在神经胶质瘤细胞和正常胶质细胞中的mRNA和蛋白质表达水平,结果发现ABCE1在神经胶质瘤细胞U251中的表达高于在正常胶质细胞中的表达。利用siRNA靶向沉默ABCE1后,神经胶质瘤细胞U251中ABCE1 mRNA和蛋白的表达水平均显著减少,细胞的凋亡率显著提高,细胞增殖和迁移明显受到抑制,而且细胞对化疗药物替莫唑胺的敏感性增强。此外,沉默ABCE1后,Bcl-2的mRNA和蛋白质表达水平显著下调,而Bax的mRNA和蛋白质表达水平显著上调。以上研究结果表明,ABCE1与神经胶质瘤细胞的增殖和迁移密切相关,通过siRNA靶向沉默ABCE1基因可显著降低U251细胞的增殖和迁移能力。     

Knockdown of VEGFR2 Inhibits Proliferation and Induces Apoptosis in Hemangioma-Derived Endothelial Cells

Angiogenesis is a process of development and growth of new capillary blood vessels from pre-existing vessels. Angiogenic growth factors play important roles in the development and maintenance of some malignancies, of which vascular endothelial growth factor (VEGF)/VEGFR2 interactions are involved in proliferation, migration, and survival of many cancer cells. The aim of this study was to investigate the function of VEGFR2 in human hemangiomas (HAs). Using immunohistochemistry assay, we examined the expression levels of VEGF, VEGFR2, Ki-67, glucose transporter-1 (Glut-1), phosphorylated protein kinase B (p-AKT) and p-ERK in different phases of human HAs. Positive expression of VEGF, VEGFR2, Ki-67, Glut-1, p-AKT and p-ERK was significantly increased in proliferating phase HAs, while decreased in involuting phase HAs (P=0.001; P=0.003). In contrast, cell apoptotic indexes were decreased in proliferating phase HAs, but increased in involuting phase HAs (P<0.01). Furthermore, we used small hairpin RNA (shRNA)-mediated VEGFR2 knockdown in primary HA-derived endothelial cells (HemECs) to understand the role of VEGF/VEGFR2 signaling. Knockdown of VEGFR2 by Lv-shVEGFR2 inhibited cell viability and induced apoptosis in primary HemECs companied with decreased expression of p-AKT, p-ERK, p-p38MAPK and Ki-67 and increased expression of caspase-3 (CAS-3); Overexpression of VEGFR2 promoted cell viability and blocked apoptosis in Lv-VEGFR2-transfected HemECs. Taken together, our findings demonstrate that, increased expression of VEGFR2 is involved in the development of primary HemECs possibly through regulation of the AKT and ERK pathways, suggesting that VEGFR2 may be a potential therapeutic target for HAs.Key words: vascular endothelial growth factor receptor 2, hemangioma, proliferation, apoptosis     

Expression of 1N3R-Tau Isoform Inhibits Cell Proliferation by Inducing S Phase Arrest in N2a Cells

Tau is a microtubule-associated protein implicated in neurodegenerative tauopathies. Six tau isoforms are generated from a single gene through alternative splicing of exons 2, 3 and 10 in human brain. Differential expression of tau isoforms has been detected in different brain areas, during neurodevelopment and in neurodegenerative disorders. However, the biological significance of different tau isoforms is not clear. Here, we investigated the individual effect of six different isoforms of tau on cell proliferation and the possible mechanisms by transient expression of eGFP-labeled tau isoform plasmid in N2a cells. Our study showed the transfection efficiency was comparable between different isoforms of tau by examining GFP expression. Compared with other isoforms, we found expression of 1N3R-tau significantly inhibited cell proliferation by Cell Counting Kit-8 assay and BrdU incorporation. Flow cytometry analysis further showed expression of 1N3R-tau induced S phase arrest. Compared with the longest isoform of tau, expression of 1N3R-tau induced cyclin E translocation from the nuclei to cytoplasm, while it did not change the level of cell cycle checkpoint proteins. These data indicate that 1N3R-tau inhibits cell proliferation through inducing S phase arrest.     

Schlafen 1 Inhibits the Proliferation and Tube Formation of Endothelial Progenitor Cells

Endothelial progenitor cells (EPCs) are the major source of cells that restore the endothelium during reendothelialization. This study was designed to investigate whether Schlafen 1 (Slfn1) has an effect on the proliferation and tube formation of EPCs in vivo. Slfn1 was expressed in rat EPCs. The overexpression of Slfn1 suppressed the proliferation and tube formation of EPCs; conversely, the knockdown of Slfn1 by shRNA promoted the proliferation and tube formation of EPCs. Furthermore, when Slfn1 was overexpressed, the EPCs were arrested in the G1 phase of the cell cycle. In contrast, when Slfn1 was knocked down, the EPCs progressed into the S phase of the cell cycle. Additionally, the overexpression of Slfn1 decreased the expression of Cyclin D1, whereas the knockdown of Slfn1 increased the expression of Cyclin D1; these findings suggest that Cyclin D1 is downstream of Slfn1 in Slfn1-mediated EPC proliferation. Taken together, these results indicate a key role for Slfn1 in the regulation of EPC biological behavior, which may provide a new target for the use of EPCs during reendothelialization.     

蛇床子素通过促进胃癌细胞N87凋亡和细胞周期阻滞而抑制细胞增殖

蛇床子素是从伞形科植物蛇床中提取的一类具有生物活性的化合物。研究显示,蛇床子素对多种肿瘤细胞具有抑制作用,然而尚未有研究揭示其对胃癌N87细胞的抗肿瘤活性。本文研究了蛇床子素在体外和荷瘤小鼠体内对胃癌N87细胞的抗肿瘤效应,并进一步利用流式细胞术、TUNEL试验及Western印迹检测分析其对细胞周期及细胞凋亡的影响,以探索其作用机制。研究结果表明,蛇床子素有效地抑制了体外培养的N87细胞生长,并呈浓度依赖效应。本文还建立了N87的荷瘤小鼠模型。结果显示,无论是在低剂量(50 mg/kg)或高剂量(100 mg/kg)情况下,蛇床子素均显示了有效的肿瘤生长抑制效果。流式细胞术及Western印迹的结果表明,蛇床子素诱导N87细胞阻滞在G_2/M期。通过流式细胞术、TUNEL测试及Western印迹结果证明,蛇床子素通过激活胱天蛋白酶-3依赖的凋亡通路,最终导致了N87细胞凋亡的发生。综上所述,本研究显示,蛇床子素在胃癌N87细胞中通过促进细胞凋亡而发挥其抗肿瘤活性,这将为其应用于胃癌的临床治疗提供理论参考。     

Baohuoside I Inhibits the Proliferation of Hepatocellular Carcinoma Cells via Apoptosis Signaling and NF-kB Pathway

Baohuoside I is a flavonoid isolated from Epimedium koreanum Nakai and has many pharmacological activities. However, its role in liver cancer remains unclear. This study aimed to investigate the inhibitory effect of Baohuoside I on the Human Hepatocellular Carcinoma (HCC) cell lines QGY7703, and underlying mechanisms. QGY7703 cells were used as the model to assess the function of Baohuoside I in vitro. The effects of Baohuoside I on QGY7703 cells’ growth, proliferation, and invasiveness were confirmed by CCK-8, lactate dehydrogenase release, and invasion assays. Cell apoptosis was analyzed by flow cytometry, and the levels of cleaved Caspase-3, Bax, and Bcl-2 were quantified by western blot. Western blot analysis, nuclear translocation of NF-κB, and Q-PCR were used to measure the expression of affected molecules. In QGY7703 cells, Baohuoside I induced the expression of molecules related to NF-κB pathway. The toxicity of Baohuoside I on QGY7703 cells was also confirmed in vivo, in a tumor model. Baohuoside I had a significant toxic effect on QGY7703 cells from a concentration of 10 μM. This compound significantly inhibited the proliferation of QGY7703 cells by inducing apoptosis and downregulating NF-κB signaling pathway. Thus, Baohuoside I is a novel candidate drug and opens new possibilities of clinical strategies for HCC treatment.