大鼠胰腺次全切除后导管细胞同源盒基因-1表达规律研究

目的 研究大鼠胰腺大部分切除术后,残余胰腺导管上皮细胞转录因子胰-十二指肠同源盒基因(PDX)-1表达规律,探讨PDX-1表达的意义。方法手术切除大鼠胰腺约90％组织,不同时间点处死动物后取材,解剖显微镜下游离胰腺导管,逆转录-聚合酶链反应(RT-PCR)与Western blot检测导管上皮中PDX-1 mRNA和蛋白表达。结果实验组手术后第2天与第3天导管上皮细胞PDX-1蛋白表达显著升高,高于对照组两倍以上,与对照组比较差异有显著性(P<0.05),第5天到第7天逐渐恢复到对照组水平;实验组在各时间点PDX-1 mRNA表达水平与对照组无明显差异(P>0.05)。结论残余胰腺再生过程中导管上皮细胞中出现PDX-1高表达,表明胰腺干细胞参与残余胰腺再生,且PDX-1表达受转录后调控。     

缺氧微环境对胰腺癌细胞发生上皮向间叶转化的诱导作用

Objective To investigate whether hypoxic environment can promote the metastasis of pancreatic cancer cells by inducing epithelial to mesenchymal transition (EMT). Methods The Panc-1 cells were cultured in hypoxia environment. After cultured for indicated periods, the in vitro invasive ability of Pane-1 cells was compared with normoxia group using Transwell invasion assay. The epithelial marker E-cadherin and the mesenchymal marker vimentin were assayed by Western blot. The expression of Snail, a strong activator of EMT, was detected by real-time PCR. The HIF-1 α encoding cDNA was transiently trans-fected into the Pane-1 cells, and the E-cadherin and vimentin were analyzed. Results The number of cells invading to the lower side of the membrane under hypoxia was ( 121±5 ), whereas only ( 84±3 ) in no-morxia group ( P < 0.05 ). The relative expression of E-cadherin in normoxia, hyoxia groups at 12,24,48 hwas (0.59±0.04 vs 54.00±0.05,0.45±0.10,0.36±0.03 ), and that of vimentin was (0.36±0.05, 0.41±0.04,0.48±0.06,0.58±0.05 ) ( P < 0.05 ). The relative expression of Snail mRNA was in-creased in hypoxia environment, and the difference was significant after 72 h ( P < 0.05 ). Before and after HIF-1α cDNA was transfected into the Panc-1 cells, the relative expression of E-cadherin was 0.63± 0.05, and 0.47±0.07, and that of vimentin was 0.47±0.07, and 0.32±0.04 respectively ( P < 0.05 ). Conclusion Hypoxia microenvironment can activate HIF-1α and Snail to trigger EMT of pancreatic cancer cells.     

大鼠急性胰腺炎严重程度与细胞因子表达水平的相关性研究

我们通过对大鼠急性胰腺炎时外周血中肿瘤坏死因子 α(ＴＮＦ α)、白细胞介素 (ＩＬ) 6、ＩＬ 10表达水平进行动态观察 ,探讨病情严重程度与以上细胞因子的相关性 ,现报道如下。一、材料和方法1.动物分组与模型制备 :ＳＤ大鼠 60只 ,体重 2 2 0～ 380 ｇ ,雌雄不限。随机分成 3、6、12、2 4ｈ组 (各 15只 ) ,每组内再分为实验组 (各 12只 ) ,正常对照组 (各 3只 )。实验前 12ｈ禁食不禁饮 ,按Ａｈｏ等[1] 的方法建立急性胰腺炎模型。自胰胆管按 0 .2ｍｌ/ｍｉｎ速度推注 5 %牛磺胆酸钠 1ｍｌ/ｋｇ鼠重。对照组术中仅刺激胰胆管肝门端而未…     

畸胎瘤细胞源性生长因子shRNA增强胰腺癌细胞株Panc-1对健择敏感性的研究

目的 通过转染针对畸胎瘤细胞源性生长因子(PCDGF)shRNA至胰腺癌细胞株Panc-1中,观察对健择敏感性的变化,分析PCDGF因子对化疗药物敏感性的影响.方法 免疫组织化学和定量聚合酶链反应(PCR)测定PCDGF在胰腺癌组织中的表达,构建含2条shRNA的RNAi质粒,脂质体转染PCDGF shRNA至Panc-1,测定转染前后细胞对健择敏感性的变化.结果 PCDGF表达于多数胰腺癌细胞的细胞质中(阳性率为84.5%),显示胰腺癌组织PCDGF的表达明显高于胰腺囊腺瘤和胰腺炎(t=11.41,P<0.05),构建PCDGF shRNA质粒经脂质体转染后PC-DGF的表达显著受到抑制(抑制率为82.3%),在较高浓度时,健择的细胞毒作用明显增强.结论 针对PCDGF的shRNA能显著增加Panc-1对健择的敏感性.     

重组组织因子途径抑制物-2对人胰腺癌细胞体外侵袭能力的影响

我们通过脂质体法将组织因子途径抑制物-2（TFPI-2）基因转染人胰腺癌细胞株Pane-1，观察其对胰腺癌细胞体外侵袭、转移能力的影响。     

乌司他丁对重症急性胰腺炎的疗效及其作用机制的研究

重症急性胰腺炎是一个有多种炎症因子如细胞因子、血小板活化因子(PAF)、一氧化氮(NO)、氧自由基(OFR)、磷脂酶A2等参与的复杂病理生理过程,炎症因子之间相互作用引发急性全身炎性反应综合征(SIRS),导致多器官功能衰竭(MOSF)[1]。本实验采用牛磺胆酸钠诱发大鼠重症急性胰腺炎合并MOSF,观察血清中TNF-α、NO及OFR的改变,同时观察乌司他丁对TNF-α、NO及OFR的影响及其治疗作用,并探讨其作用机理。材料与方法一、动物分组:Wistar大鼠70只(同济医科大学动物实验中心提供),250～300克,雌雄各半,随机分为4组:(1)正常组:10只,剖腹…     

一氧化氮对重症急性胰腺炎肺损伤的作用及其机制

目的研究一氧化氮(NO)减轻重症急性胰腺炎(SAP)肺损伤的机制。方法采用逆行胰胆管牛磺胆酸钠注射制造SAP大鼠模型。动物分为假手术组、胰腺炎组、L-精氨酸(L-Arg)治疗组和氯喹(CQ)治疗组。硝酸还原酶法检测肺组织NO的浓度变化,实时定量PCR(RT-PCR)检测肺组织TLR(Toll-like receptor)2／4 mRNA表达变化。结果与假手术组比较,SAP大鼠肺组织NO浓度降低,肺损伤加重;肺组织TLR2／4 mRNA表达增高,肺组织肿瘤坏死因子-α(TNF-α)表达升高(P<0．05)。给予不同剂量L-Arg治疗后,肺组织NO浓度明显升高,肺损伤程度减轻;TLR2／4 mRNA表达降低,肺组织TNF-α表达降低(P<0．05)。给予CQ抑制肺组织TLR2／4 mRNA表达后,肺组织内NO浓度升高,肺损伤减轻,TNF-α表达降低(P<0．05)。结论NO可以明显抑制SAP肺组织TLR2／4 mRNA的表达,减少细胞因子的合成及释放,从而减轻肺组织损伤。     

《急性胰腺炎诊治指南(2014版)》热点问题解读

近年来,急性胰腺炎(AP)的研究取得了巨大进展,其诊治的很多重要方面发生了明显的变化.2014年,中华医学会外科学分会胰腺外科学组对我国2007年发布的《重症急性胰腺炎诊治指南》进行了修订.修订后的指南,按严重度将AP分为轻症急性胰腺炎(MAP)、中重症急性胰腺炎(MSAP)和重症急性胰腺炎(SAP)三类,后两者主要区别是MSAP器官功能衰竭持续的时间≤48 h,而SAP＞ 48 h.影像学评估采用改良的CT严重指数(MSCTI)评分.局部并发症包括急性胰周液体积聚(APFC)、急性坏死物积聚(ANC)、包裹性坏死(WON)和假性囊肿.病程分期为早期(急性期)、中期(演进期)和晚期(感染期).感染性坏死是手术指征,无症状的无菌性坏死无需手术.手术须遵循延迟原则.手术方式包括经皮穿刺引流(PCD)、微创手术及开放手术,这些术式可单独或联合应用.     

缺氧微环境对胰腺癌细胞发生上皮向间叶转化的诱导作用

Objective To investigate whether hypoxic environment can promote the metastasis of pancreatic cancer cells by inducing epithelial to mesenchymal transition (EMT). Methods The Panc-1 cells were cultured in hypoxia environment. After cultured for indicated periods, the in vitro invasive ability of Pane-1 cells was compared with normoxia group using Transwell invasion assay. The epithelial marker E-cadherin and the mesenchymal marker vimentin were assayed by Western blot. The expression of Snail, a strong activator of EMT, was detected by real-time PCR. The HIF-1 α encoding cDNA was transiently trans-fected into the Pane-1 cells, and the E-cadherin and vimentin were analyzed. Results The number of cells invading to the lower side of the membrane under hypoxia was ( 121±5 ), whereas only ( 84±3 ) in no-morxia group ( P < 0.05 ). The relative expression of E-cadherin in normoxia, hyoxia groups at 12,24,48 hwas (0.59±0.04 vs 54.00±0.05,0.45±0.10,0.36±0.03 ), and that of vimentin was (0.36±0.05, 0.41±0.04,0.48±0.06,0.58±0.05 ) ( P < 0.05 ). The relative expression of Snail mRNA was in-creased in hypoxia environment, and the difference was significant after 72 h ( P < 0.05 ). Before and after HIF-1α cDNA was transfected into the Panc-1 cells, the relative expression of E-cadherin was 0.63± 0.05, and 0.47±0.07, and that of vimentin was 0.47±0.07, and 0.32±0.04 respectively ( P < 0.05 ). Conclusion Hypoxia microenvironment can activate HIF-1α and Snail to trigger EMT of pancreatic cancer cells.