电离辐射诱导HIF-1α-Survivin通路在人淋巴瘤细胞中的活化

目的 研究电离辐射对人非霍奇金淋巴瘤细胞中HIF-1α-Survivin通路活化状态的影响,探讨恶性淋巴瘤放射抗性的机制.方法 采用Western blot方法检测辐射后3种淋巴瘤细胞中HIF-1α和Survivin蛋白的表达水平,观察应用HIF-1α抑制剂Echinomycin及转染反义HIF-1a siRNA对Survivin蛋白及mRNA表达的影响.结果 在人非霍奇金淋巴瘤细胞中存在HIF-1α和Survivin蛋白的表达,X射线照射后10～20 h出现HIF～1α蛋白表达增加,照射后24 h Survivin蛋白表达增加,与对照组比较差异有统计学意义(t =7.53 ～31.31,P＜0.01).与单纯照射组比较,采用HIF-1α抑制剂预处理肿瘤细胞后,Survivin蛋白表达下降,且具有药物浓度依赖性(t=7.21 ～32.81,P＜0.01).而转染反义HIF-1α siRNA后,照射未诱导产生survivin mRNA和蛋白表达增加.结论 电离辐射能够活化人非霍奇金淋巴瘤细胞中的HIF-1α-Survivin通路,可能与肿瘤的放射抗性有关.     

MEBT/ MEBO对慢性难愈合创面 NF-κB p65、IκBα、IKK及其磷酸化蛋白表达水平的影响

【摘要】 目的 探讨创疡再生医疗技术 (MEBT/MEBO) 对慢性难愈合创面组织中核因子 κB (NF-κB) p65?NF-κB 抑制蛋白(IκB)、IκB激酶(IKK) 及其磷酸化蛋白表达水平的影响。 方法 选取 SPF 级雄性 Wistar 大鼠90 只适应性饲养1周后,按照随机数表法将其随机分为空白组、对照组、模型组、MEBO组和贝复新组,每组18 只。空白组大鼠仅做备皮处理,对照组大鼠建立急性创面模型,模型组、MEBO组、贝复新组大鼠建立慢性难愈合创面模型?空白组大鼠备皮处皮肤及对照组、模型组大鼠创面采用生理盐水纱布换药处理,MEBO 组大鼠创面采用湿润烧伤膏 (MEBO) 药纱换药处理,贝复新组大鼠创面采用重组牛碱性成纤维细胞生长因子换药处理,对比各组大鼠干预第3、7、14天创面愈合率、组织病理学变化,以及皮肤/ 创面组织中NF-κB p65、IκBα、IKK及 p-NF-κB p65、p-IκBα、p-IKK 蛋白表达水平。结果 (1)干预第 3、7 天,各组大鼠创面愈合率均无明显差异(P均＞0. 05);干预第 14 天,模型组大鼠创面愈合率明显低于对照组、MEBO组(P均＜0.05),其余各组间创面愈合率均无明显差异 (P均＞0.05)。(2)干预第 7 天,MEBO 组、贝复新组大鼠创面组织中胶原纤维量均明显增加,可见大量成纤维细胞和新生毛细血管及少量毛囊结构生成; 干预第14天,MEBO 组、贝复新组大鼠创面组织中胶原纤维量明显多于模型组,可见整齐排列的毛细血管及成熟的毛囊、皮脂腺等组织,而模型组大鼠创面组织中仍可见少量炎症细胞浸润,但已有大量成纤维细胞生成。(3) 干预第7、14天,空白组、对照组、MEBO组大鼠创面组织中 NF-κB p65、IKK、p-NF-κB p65、p-IKK、p-IκBα 蛋白表达水平均明显低于模型组(P均＜0.05);干预第 7 天,MEBO 组大鼠创面组织中 NF-κB p65、IκBα 蛋白表达水平均明显高于贝复新组 (P均＜0.05),IKK、p-NF-κB p65、p-IKK、p-IκBα 蛋白表达水平均明显低于贝复新组 (P 均＜0.05); 干预第 14天,MEBO 组大鼠创面组织中 IKK、IκBα蛋白表达水平均明显低于贝复新组 (P 均＜0.05)。结论 MEBT/ MEBO促进慢性难愈合创面愈合的机制可能与创面组织中 NF-κB p65、IκBα、IKK 及其磷酸化蛋白的表达水平有关。     

电离辐射对小鼠脾细胞中调节性T细胞及其相关分子表达的影响

Objective To observe the changes in expressions of spleen regulatory T cells (Tregs)and the related factor forkhead box protein-3 (Foxp3) after irradiation with different doses of X-ray in mice at different times,and to elaborate the effects of X-rays on regulatory T cells and Foxp3.Methods 112male ICR mice were randomly divided into 2 groups and irradiated by X-rays at the doses of 0.075 and 2 Gy,respectively.The mice were killed at 0,4,8,16,24,48,and 72 h post-irradiation and the spleens removed.Flow cytometry was used to detect the percentage of CD4 + CD25 + Treg and protein expression of (Foxp3),and RT-PCR was used to exmiamine the mRNA expression of Fox3.Results Compared with those before irradiation,the CD4 + CD25 + Treg positive rates began to increase and peaked at 8 h post-irradiation with 0.075 Gy at 8,16,24,72 h(t = 8.73,10.55,4.21,4.65 ,P ＜ 0.05) and 2 Gy at 8,16,48,72 h(t = 4.65,4.28,3.71,2.88,P ＜ 0.05),and then slightly decreased,but still remained at high levels.The mRNA protein levels of Fox3 did not change significantly after exposure to the dose of 0.075 Gy,but began to significantly increase at 8 h after exposure to the dose of 2 Gy.However,the Foxp3 protein level began to increase 4 h post-irradiation,peaked at 16 h,and then slightly decreased,but still ramained at high levels (t =2.59,3.37,3.70,3.20,P＜0.05).Conclusions The changes in expressions of Tregs and Foxp3 after high- and low-dose X-ray irradiation may be used to explain the differences in immune effects induced by ionizing radiation at different doses.     

X射线对人肺癌A549细胞CCR7表达影响的研究

目的 研究不同剂量X射线照射及照射后不同时间点对人肺癌A549细胞CC-趋化因子受体7(CCR7)表达的影响.方法 体外培养A549细胞,实验组采用直线加速器X射线一次性照射,细胞吸收剂量分别为2、4、6和8 Gy(源皮距100 cm;剂量率442.89 cGy/min),照射后4、12、24、48和72 h分别采用实时荧光定量PCR技术及Western blot方法分别进行CCR7 mRNA及蛋白质表达水平检测;对照组A549细胞除不接受x射线照射外,余处理同实验组.结果 A549细胞经2、4、6和8 Gy的X射线照射后,CCR7 mRNA及蛋白质在照射4 h后开始表达升高,达到高峰后相继出现下降;72 h后6和8 Gy组mRNA表达量仍高于对照组水平(t=6.75～7.26,P＜0.01),2和6 Gy组蛋白质表达量高于对照组(t=11.13～14.17,P＜0.01),而4和8 Gy组蛋白质表达量在48和72 h已降至对照组水平.结论 2、4、6和8 Gy的X射线照射A549细胞后,A549细胞CCR7mRNA及蛋白质的表达量明显增加,可能与一定剂量X射线辐射促进A549细胞增殖和转移有关.

Abstract：

Objective To study the effects of X-ray radiation on CC-chemokine receptor 7(CCR7) expression in human non-small cell lung cancer (NSCLC) cells.Methods Humanadenocarcinoma cells of the line A549 were cultured and irradiated by X-ray at the absorbed doses of 2,4,6,and 8 Gy respectively by linear accelerator (with the source skin distance of 100 cm and dose rate of 442.89 cGy/min).The relative levels of CCR7 mRNA and protein expression in the A549 cells were respectively detected by real time-PCR and Western blotting 4,12,24,48,and 72 h after radiation.Untreated A549 cells were used as control group.Results The expression levels of CCR7 mRNA and protein in the A549 cells began to increase since 4 h after radiation and then decreased gradually after they reached the peak.The CCR7 mRNA expression levels 72 h after radiation of the 6 and 8 Gy groups were still significantly higher than those of the control group (t = 6.75-7.26,both P < 0.01),and the CCR7 protein expression levels of the 2 and 6 Gy group were still significantly higher than those of the control group(t=11.13-14.17,both P <0.01).Then the CCR7 protein expression levels of the 4 and 8 Gy groups decreased to the control group level 48 and 72 h after radiation respectively.Conclusions The CCR7 mRNA and protein expression levels in the NSCLC cells increase after X-ray irradiation,which may be correlated with the promotion of proliferation and metastasis of NSCLC cells by X-ray irradiation at a certain dose.     

慢病毒载体介导HIF-1α RNAi对人胰腺癌细胞Patu8988相关基因表达的影响

目的 探讨慢病毒表达载体介导HIF-1α RNA干扰(RNAi)对人胰腺癌细胞Patu8988HIF-1α和Glut-1表达的影响.方法 构建针对HIF-1α基因的RNAi慢病毒表达载体LV-RNAi-HIF-1α.乏氧条件下体外培养4h,转染LV-RNAi-HIF-1α的Patu8988细胞为实验组;转染空病毒载体和未转染任何病毒载体Patu8988细胞分别为阴性对照组和空白对照组.采用荧光定量RT-PCR和Western印迹法检测Patu8988细胞HIF-1α表达情况,采用RT-PCR法检测转染LV-RNAi-HIF-1α后Patu8988细胞Glut-1的表达情况.采用SPSS 17.0软件行单因素方差分析和两样本t检验分析各组之间的差异.结果 构建的慢病毒表达载体LV-RNAi-HIF-1α,在常氧和乏氧状态下致HIF-1α mRNA表达分别下降65.1％(0.209/0.321)和80.6％ (0.791/0.982)(t=10.52和15.24,均P＜0.05),阴性对照组分别为0.6％(0.002/0.321)和7.2％(0.071/0.982)(t=5.26和7.38,均P＜0.05);乏氧状态下实验组、阴性对照组和空白对照组HIF-1α蛋白的表达分别为0.159±0.010、0.745±0.012和0.711±0.023,差异有统计学意义(F=35.52,t=6.72和10.56,均P＜0.05),实验组较其他2组表达下降;乏氧条件下实验组Glut-1 mRNA表达(0.040±0.003)较阴性对照组(0.054±0.003)和空白对照组(0.062±0.004)均有明显下降(F=35.28,t=5.94和8.55,均P＜0.01).结论 通过构建LV-RNAi-HIF-1α慢病毒表达载体沉默HIF-1α基因表达,可降低Patu8988胰腺癌细胞Glut-1 mRNA表达.     

辐射对Jurkat细胞P21蛋白及ICR小鼠胸腺细胞p21基因表达的影响

目的 探讨电离辐射对Jurkat细胞P21蛋白和ICR小鼠胸腺细胞p21基因表达的影响.方法 采用流式细胞术(FCM),检测0、0.5、1.0、2.0、4.0及6.0 Gy X射线照射后Jurkat细胞中P21蛋白表达的变化.采用实时定量PCR技术,分别检测0、0.5、1.0、2.0、4.0及6.0 Gy X射线照射后4和24 hd'鼠胸腺及脾细胞中p21基因表达的变化.结果 不同剂量X射线照射后12和24 h,Jurkat细胞中P21蛋白表达在0.5～4.0 Gy范围内均随剂量的增大而升高(t=-24.23～-3.96,P<0.05),6 Gy时均出现表达下降(t=-11.19、-14.50,P<0.05);与假照射组相比,在0～6.0 Gy照射后4和24 h,小鼠胸腺及脾细胞中p21基因的相对表达量均随剂量增大逐渐增加(t=-29.96～8.80,P<0.05);并于6.0 Gy时达最高(t=-11.84～-3.42,P<0.05),仅胸腺细胞1 Gy照射后4 h除外(t=-3.42,P>0.05).结论 x射线能诱导P21蛋白及基因表达增加,并在一定剂量范围内存在良好的剂量-效应关系.

Abstract：

Objective To investigate the effects of ionizing radiation on the expression of P21 protein in Jurkat cell line and p21 gene in thymocytes and splenocytes of mice.Methods Flow cytometry (FCM)was used to analyze the expression of P21 protein in Jurkat cells at 12 and 24 h after irradiation to 0,0.5,1.0,2.0,4.0,and 6.0 Gy.Real-time PCR was used to detect the expression of p21 gene in thymocytes and splenocytes of mice at4 and 24 h after irradiation to 0,0.5,1.0,2.0,4.0,and 6.0 Gy.Multi-staining was used to analyze the micronucleus rates of Rct in bone marrow.Results The expressions of P21 protein were increased in a dose-dependent manner during 0.5-4.0 Gy(t=-24.23--3.96,P<0.05),but decreased at 6.0 Gy at 12 and 24 h post-irradiation(t=-11.19,-14.50,P<0.05).The expressions of p2 1 gene in both thymocytes and splenocytes of mice were increased in dose-dependent manner in the range of 0-6.0 Gy(including 6.0 Gy)(t=-29.96-8.80,P<0.05),and reached to the peak at 6.0 Gy at 4 and 24 h post-irradiation(t=-11.84--3.42,P<0.05),except thymocytes at 4 h and 1.0 Gy post-irradiation(t=-3.42,P>0.05).Conclusions The expressions of P21 protein and p21 gene could be increased by X-ray irradiation.which shows good dosedependent manners in certain range of dose.     

NF-kB抑制与X射线诱导的人非霍奇金淋巴瘤细胞凋亡的研究

目的 探讨NF-kB p65对X射线诱导人非霍奇金淋巴瘤(non-Hodgkin lymphoma,NHL)细胞凋亡的作用及其调控机制.方法 以NF-kB p65抑制剂quinazoline (QNZ)处理人NHL细胞.将NHL细胞株Namalwa、Ramos和Raji细胞分为空白对照组、单纯照射组(IR)和X射线+QNZ实验组( IR+ QNZ),采用Annexin-V染色方法检测肿瘤细胞凋亡水平的变化,采用Western blot方法检测各细胞株中Survivin及凋亡相关蛋白Bax、Bcl-2和Cleaved Caspase-3的表达水平;应用实时定量PCR方法检测Survivin mRNA水平.结果 应用流式细胞仪检测凋亡细胞百分比,结果显示,QNZ处理后进行照射与单纯照射组相比凋亡细胞明显增加,且具有药物浓度依赖性(t=2.93～12.52,P＜0.05),Western blot法检测结果显示,电离辐射可显著增加人NHL细胞中Survivin蛋白的表达.而应用1、10和50 nmol/L QNZ处理人NHL细胞24 h后进行照射,可显著下调电离辐射所诱导的NHL细胞中的Survivin蛋白表达.随药物浓度增加其作用更为明显(t=3.29～ 16.72,P＜0.05).同时凋亡相关蛋白Bcl-2表达减低,而Bax和Cleaved caspase-3蛋白表达增加,Bcl-2/Bax比值明显降低,与单纯照射组相比,差异有统计学意义(t =6.20 ～9.91,P＜0.05).预处理QNZ后射线诱导的3种NHL细胞Survivin mRNA均不同程度下降.结论 抑制NF-kB能够增加X射线诱导的NHL细胞凋亡,其机制可能与下调Survivin蛋白表达水平及对凋亡相关蛋白Bcl-2家族的调节有关.     

NF-κB抑制剂对烫伤脓毒症大鼠致炎/抗炎细胞因子表达的影响

目的观察NF-κB抑制剂对动物主要器官致炎/抗炎细胞因子基因及蛋白表达的调节效应,并对其作用机制进行初步探讨.方法34只动物随机分为正常对照组(n=6)、烫伤(20%TBSAⅢ度)对照组(n=6)、烫伤后金葡菌感染组(n=12)和NF-κB抑制剂二硫氨基甲酸酞吡咯烷(PDTC)拮抗组(n=10),检测动物肝、肾、肺组织中TNF-α、IL-10基因及蛋白表达的改变.结果烫伤脓毒症组0.5～2h肝、肺、肾组织中TNF-α mRNA表达迅速增强,同时各组织TNF-α蛋白水平亦显著升高.PDTC早期干预对肺脏TNF-α mRNA及蛋白水平影响不明显,但肝、肾组织其表达量在2h均被显著抑制(P＜0.05或0.01).烫伤合并金葡菌感染后2h大鼠肝、肺、肾组织IL-10 mRNA与蛋白水平均显著升高(P＜0.05或0.01),早期给予PDTC对肺、肾组织各时间点IL-10 mRNA与蛋白表达均无明显影响,肝组织仅2h呈现一定程度地下降.结论NF-κB抑制剂在有效拮抗G+菌脓毒症TNF-α等致炎介质的同时,对机体并存的抗炎细胞因子反应机制具有保护作用.     

缺氧诱导因子1α在异丙肾上腺素诱导的大鼠心房纤维化中的表达及意义

目的 探讨缺氧诱导因子1α(HIF-1α)在盐酸异丙肾上腺素(ISO)诱导的大鼠心房纤维化中的表达及其可能机制.方法 健康雄性Wistar大鼠30只,随机均分为空白对照组、ISO组、ISO+西罗莫司(Rapa)干预组(Rapa组).于实验15d时处死大鼠取心肌组织,放射免疫法检测血管紧张素Ⅱ(AngⅡ)的含量,HE和Masson染色法观察纤维化程度即胶原容积分数(CVF),免疫组织化学法、Western blotting检测HIF-1α、转化生长因子β1(TGF-β1)和基质金属蛋白酶9(MMP-9)在大鼠心房纤维化组织中的表达,并分析HIF-1α、TGF-β1、MMP-9蛋白表达量与心房纤维化指标CVF的相关性,以及HIF-1α、TGF-β1、MMP-9蛋白表达量之间的相关性.结果 ISO组、Rapa组AngⅡ含量较空白对照组明显升高(P＜0.01).空白对照组CVF为15.482％±0.837％,无心房纤维化,而Rapa组、ISO组CVF分别为16.730％±1.052％、86.704％±1.928％,Rapa组较ISO组的心房纤维化程度明显减弱(P＜0.01).免疫组化及Western blotting检测显示,与空白对照组相比,ISO组HIF-1α、TGF-β1和MMP-9蛋白表达量均明显增加(P＜0.01),与ISO组比较,Rapa组中HIF-1α、TGF-β1和MMP-9蛋白表达量明显降低(P＜0.01). HIF-1α、TGF-β1和MMP-9蛋白表达量均与心房纤维化程度(以CVF为评价指标)呈正相关(r=0.987,r=0.988,r=0.917,P＜0.01); HIF-1α与TGF-β1、MMP-9蛋白表达量呈正相关(r=0.976,r=0.901,P＜0.01),MMP-9与TGF-β1蛋白表达量亦呈正相关(r=0.912,P＜0.01).结论 在异丙肾上腺素诱导的大鼠心房纤维化过程中,AngⅡ、HIF-1α、TGF-β1、MMP-9发挥着重要作用.     

HIF-1α与NF-κB在妊娠期高血压疾病患者胎盘组织中的表达及意义

目的检测胎盘组织中HIF-1α和NF-κB的表达,探讨其与妊娠期高血压疾病发生的关系。方法选择2006年12月—2007年12月在我院住院分娩的孕妇50例,分为妊娠期高血压组、轻度子痫前期组、重度子痫前期组、子痫组、正常妊娠组,采用免疫组化法检测50例胎盘标本绒毛上皮组织中HIF-1α和NF-κB的表达。结果(1)妊娠期高血压疾病组HIF-1α与NF-κB表达较正常妊娠组明显升高,且差异有统计学意义(P<0.05);(2)妊娠期高血压疾病组胎盘HIF-1α与NF-κB的表达呈正相关(P<0.01),而正常妊娠组相关性不明显(P>0.05)。结论胎盘组织中HIF-1α与NF-κB的表达升高与妊娠期高血压疾病的发生有关。     

缺氧诱导体外绒毛组织转化生长因子3β表达及调控

 目的探讨低氧对体外培养的绒毛组织转化生长因子3β(TGF-3β)表达的诱导作用,以及缺氧诱导因子1(HIF-1)对TGF-3β表达的调控作用.方法取孕早期绒毛组织分4组培养:正常对照组、缺氧组、HIF-1α反义组和HIF-1α正义组.HIF-1α反义组和HIF-1α正义组先加入HIF-1α寡核甘酸,低氧条件培养48h.48h后,收集培养的绒毛组织,实时定量PCR方法检测TGF-3β和HIF-1α mRNA表达水平;Westem blot方法检测TGF-3β和HIF-1α蛋白表达水平.结果与正常对照组相比,缺氧组的HIF-1α mRNA和蛋白表达增加,TGF-3β mRNA和蛋白表达也升高;与HIF-1α正义组比较,HIF-1α反义组HIF-1α mRNA和蛋白表达降低,TGF-3βmRNA和蛋白表达也下降.结论缺氧可以诱导体外培养的绒毛组织TGF-3β表达,且缺氧对TGF-3β表达的诱导作用可能通过HIF-1调控.     

低氧诱导肾细胞癌786-O细胞HIF-1α的表达变化及意义

目的探讨缺氧条件下肾透明细胞癌786-O细胞缺氧诱导因子1α（HIF-1α）的表达变化及意义。方法786-O细胞在缺氧条件下培养不同时间（0、8、16、24h）后,RT-PCR法检测HIF-1α mRNA的转录情况;Western blot法检测HIF-1α蛋白表达变化。结果常氧状态下HIF-1α mRNA和蛋白均有一定表达。缺氧处理一定时间（8、16、24h）后786-O细胞HIF-1α mRNA表达没有明显改变（P〉0．05）,而HIF-1α蛋白表达水平显著升高（P〈0．01）。结论缺氧条件下786-O细胞HIF-1α表达变化主要在翻译水平而非转录水平。在蛋白水平对HIF-1α表达进行调控可作为肾透明细胞癌治疗的新靶点。     

三氧化二砷通过抑制IKK/NF-κB信号通路活化发挥促乳腺癌细胞凋亡效应

目的探讨IKK/NF-κB信号通路在三氧化二砷（As2O3）诱导乳腺癌细胞凋亡反应中的作用。方法以乳腺癌细胞MCF7为靶细胞,以As2O3为刺激源,锥虫蓝（台盼蓝）拒染方法检测死亡细胞比率;双荧光素酶报告基因法检测MCF7细胞中NF-κB的活化状态;Western印迹方法检测IKK/NF-κB途径各信号分子的表达水平和活化状态;RT-PCR方法检测IKK/NF-κB途径下游靶基因的表达水平。结果 As2O3可显著诱导MCF7细胞凋亡;同时NF-κB的转录活化水平及其下游凋亡反应相关靶基因的表达水平也明显下降。在此过程中,I-κB的表达水平和NF-κB关键组成亚基（p65、p50）的核浆分布状态没有改变,但IKK激酶的两个催化亚基IKKα和IKKβ的表达水平明显下调。一过性高表达IKKα和IKKβ不仅能够恢复NF-κB的活化状态,而且能够拮抗As2O3诱导的乳腺癌细胞凋亡反应。结论 As2O3可通过在蛋白激酶水平抑制IKK/NF-κB信号通路活化从而发挥促乳腺癌细胞凋亡效应。     

三氧化二砷通过抑制IKK/NF-κB信号通路活化发挥促乳腺癌细胞凋亡效应

目的探讨IKK/NF-κB信号通路在三氧化二砷(As2O3)诱导乳腺癌细胞凋亡反应中的作用。方法以乳腺癌细胞MCF7为靶细胞,以As2O3为刺激源,锥虫蓝(台盼蓝)拒染方法检测死亡细胞比率;双荧光素酶报告基因法检测MCF7细胞中NF-κB的活化状态;Western印迹方法检测IKK/NF-κB途径各信号分子的表达水平和活化状态;RT-PCR方法检测IKK/NF-κB途径下游靶基因的表达水平。结果 As2O3可显著诱导MCF7细胞凋亡;同时NF-κB的转录活化水平及其下游凋亡反应相关靶基因的表达水平也明显下降。在此过程中,I-κB的表达水平和NF-κB关键组成亚基(p65、p50)的核浆分布状态没有改变,但IKK激酶的两个催化亚基IKKα和IKKβ的表达水平明显下调。一过性高表达IKKα和IKKβ不仅能够恢复NF-κB的活化状态,而且能够拮抗As2O3诱导的乳腺癌细胞凋亡反应。结论 As2O3可通过在蛋白激酶水平抑制IKK/NF-κB信号通路活化从而发挥促乳腺癌细胞凋亡效应。     

NF-κB抑制与X射线诱导的人非霍奇金淋巴瘤细胞凋亡的研究

目的 探讨NF-κB p65对X射线诱导人非霍奇金淋巴瘤(non-Hodgkin lymphoma, NHL)细胞凋亡的作用及其调控机制。方法 以NF-κB p65抑制剂quinazoline(QNZ)处理人NHL细胞。将NHL细胞株Namalwa、Ramos和 Raji细胞分为空白对照组、单纯照射组(IR)和X射线+QNZ实验组(IR+QNZ),采用Annexin-Ⅴ染色方法检测肿瘤细胞凋亡水平的变化,采用Western blot方法检测各细胞株中Survivin及凋亡相关蛋白Bax、Bcl-2和Cleaved Caspase-3的表达水平;应用实时定量PCR方法检测Survivin mRNA水平。结果 应用流式细胞仪检测凋亡细胞百分比,结果显示,QNZ处理后进行照射与单纯照射组相比凋亡细胞明显增加,且具有药物浓度依赖性(t=2.93~12.52, P<0.05),Western blot法检测结果显示,电离辐射可显著增加人NHL细胞中Survivin蛋白的表达。而应用1、10和50 nmol/L QNZ处理人NHL细胞24 h后进行照射,可显著下调电离辐射所诱导的NHL细胞中的Survivin蛋白表达。随药物浓度增加其作用更为明显(t=3.29~16.72,P<0.05)。同时凋亡相关蛋白Bcl-2表达减低,而Bax和Cleaved caspase-3蛋白表达增加,Bcl-2/Bax比值明显降低,与单纯照射组相比,差异有统计学意义(t=6.20~9.91, P<0.05)。预处理QNZ后射线诱导的3种NHL 细胞Survivin mRNA均不同程度下降。结论 抑制NF-κB能够增加X射线诱导的NHL细胞凋亡,其机制可能与下调Survivin蛋白表达水平及对凋亡相关蛋白Bcl-2家族的调节有关。     

Effects of seawater immersion combined with open abdominal injury on the expression of NF-κB,and IκBαof small intestine in rats

Objective To observe effects of seawater immersion combined with open abdominal injury on the expression of NF-κB,and IκBαas well as the change pattern in rats. Methods Ninety-one Wistar rats were randomly divided into 3 groups: the control group (n =7), the open abdominal injury group(n =42) and open abdominal injury combined with 1-hour seawater immersion group ( n =42). The expression of NF-κB,andIκBαin small intestine tissues was measured by Western blot and statistical analyses were also made in the study. Results The expression of NF-κB,in the seawater immersion combined with open abdominal injury group increased significantly 3 hours after injury, when compared with that of the open abdominal injury group(P<0. 05), whereas the expression of NF-κB, of the pure injury group was slightly lower than that of the control group, but no statistical differences could be seen between them(P>0.05). The change pattern in the expression of IκBαwas quite the opposite to that of NF-κB. Conclusions NF-κB seemed to be rapidly and persistently involved in the whole inflammatory response to trauma induced by opened abdominal injury and seawater immersion, when a comparison was made with the pure open abdominal injury group. Injuries for the rats in the open abdominal injury combined with seawater immersion group were serious, and the feedback mechanism for NF-κB was not established for quite a long time.     

Effects of seawater immersion combined with open abdominal injury on the expression of NF-κB,and IκBαof small intestine in rats

Objective To observe effects of seawater immersion combined with open abdominal injury on the expression of NF-κB,and IκBαas well as the change pattern in rats. Methods Ninety-one Wistar rats were randomly divided into 3 groups: the control group (n =7), the open abdominal injury group(n =42) and open abdominal injury combined with 1-hour seawater immersion group ( n =42). The expression of NF-κB,andIκBαin small intestine tissues was measured by Western blot and statistical analyses were also made in the study. Results The expression of NF-κB,in the seawater immersion combined with open abdominal injury group increased significantly 3 hours after injury, when compared with that of the open abdominal injury group(P<0. 05), whereas the expression of NF-κB, of the pure injury group was slightly lower than that of the control group, but no statistical differences could be seen between them(P>0.05). The change pattern in the expression of IκBαwas quite the opposite to that of NF-κB. Conclusions NF-κB seemed to be rapidly and persistently involved in the whole inflammatory response to trauma induced by opened abdominal injury and seawater immersion, when a comparison was made with the pure open abdominal injury group. Injuries for the rats in the open abdominal injury combined with seawater immersion group were serious, and the feedback mechanism for NF-κB was not established for quite a long time.     

Effects of seawater immersion combined with open abdominal injury on the expression of NF-κB,and IκBαof small intestine in rats

Objective To observe effects of seawater immersion combined with open abdominal injury on the expression of NF-κB,and IκBαas well as the change pattern in rats. Methods Ninety-one Wistar rats were randomly divided into 3 groups: the control group (n =7), the open abdominal injury group(n =42) and open abdominal injury combined with 1-hour seawater immersion group ( n =42). The expression of NF-κB,andIκBαin small intestine tissues was measured by Western blot and statistical analyses were also made in the study. Results The expression of NF-κB,in the seawater immersion combined with open abdominal injury group increased significantly 3 hours after injury, when compared with that of the open abdominal injury group(P<0. 05), whereas the expression of NF-κB, of the pure injury group was slightly lower than that of the control group, but no statistical differences could be seen between them(P>0.05). The change pattern in the expression of IκBαwas quite the opposite to that of NF-κB. Conclusions NF-κB seemed to be rapidly and persistently involved in the whole inflammatory response to trauma induced by opened abdominal injury and seawater immersion, when a comparison was made with the pure open abdominal injury group. Injuries for the rats in the open abdominal injury combined with seawater immersion group were serious, and the feedback mechanism for NF-κB was not established for quite a long time.