替米沙坦、瑞舒伐他汀对胰岛素抵抗大鼠骨骼肌小窝蛋白1、葡萄糖转运蛋白4表达的影响

目的测定不同饮食喂养大鼠和替米沙坦、瑞舒伐他汀干预后大鼠产生胰岛素抵抗(IR)的情况,观察大鼠骨骼肌中小窝蛋白(Caveolin)1、葡萄糖转运蛋白(GLUT)4表达的变化。方法 4周龄Wistar雄性大鼠48只,以高糖高脂饮食诱导建立IR模型,以高糖高脂给药(替米沙坦、瑞舒伐他汀)诱导建立实验组,通过RT-PCR方法检测IR大鼠骨骼肌组织中Caveolin 1、GLUT4的表达水平。结果高糖高脂喂养组产生IR,给予替米沙坦、瑞舒伐他汀干预后IR明显改善,高糖高脂组Caveolin1 mRNA在骨骼肌中的表达明显高于对照组(P<0.05),高糖高脂组GLUT4 mR-NA在骨骼肌中的表达明显低于对照组(P<0.05),替米沙坦、瑞舒伐他汀可降低Caveolin1 mRNA在IR大鼠骨骼肌中的表达,升高GLUT4 mRNA在IR大鼠骨骼肌中的表达。结论①高糖高脂饲料喂养8 w可诱导大鼠IR。②IR大鼠骨骼肌组织Caveolin 1表达增加,GLUT4表达减少。③通过替米沙坦类、他汀类药物干预,可改善IR,降低Caveolin1表达水平,升高GLUT4表达水平。     

水通道蛋白1和4在大鼠挫伤肺组织中的表达

目的探讨水通道蛋白(AQP)1和4在老年大鼠挫伤后肺组织中的表达。方法通过自由落体模型制作老年鼠肺挫伤模型,分别于伤后1,3和6 h处死,取右侧肺上叶组织测水含量,RT-PCR法测量AQP1和4的mRNA表达,Western印迹法检测蛋白AQP1和4的表达水平。结果伤后老年大鼠的肺组织水含量比明显增加,与假手术组相比,AQP1和4的mRNA和蛋白表达在术后逐渐升高,术后6 h有显著差异(P<0.05),AQP-1的蛋白表达量亦明显升高(P<0.05)。结论 AQP1和4在挫伤后的肺组织中表达升高,靶向调节二者的表达可能对于肺挫伤后肺水肿的治疗具有重要意义。     

急性肺损伤大鼠肺水通道蛋白1和5的表达及功能的实验研究

目的观察脂多糖(LPS)、肿瘤坏死因子α(TNFα)、白细胞介素1β(IL1β)对大鼠肺微血管内皮细胞(LMECs)水通道蛋白1(AQP1)表达和功能的影响;同时在LPS诱发的大鼠急性肺损伤(ALI)模型观察AQP1、AQP5表达的变化。方法(1)体外实验:将第3代LMECs随机分为LPS组、TNFα组、IL1β组和DMEM对照组,实验组分别给予LPS、TNFα、IL1β刺激,应用逆转录聚合酶链反应(RTPCR)测定AQP1mRNA的表达以及免疫组化测定AQP1蛋白表达,同时采用放射性核素示踪法测定LMECs内氚水(3H2O)的放射强度。(2)体内实验:40只雄性Wistar大鼠随机分为LPS2h组、LPS4h组、LPS6h组、LPS8h组和对照组,LPS各组制成ALI模型,采用RTPCR测定ALI大鼠AQP1、AQP5mRNA表达以及免疫组化观察AQP1、AQP5蛋白表达。结果(1)体外实验:LPS、TNFα、IL1β组LMECsAQP1mRNA和蛋白表达显著低于DMEM对照组(mRNA表达分别为0.428±0.026、0.446±0.029、0.454±0.023和0.793±0.035,蛋白表达分别为0.366±0.009、0.374±0.014、0.377±0.007和0.660±0.013,P均<0.01);且LMECs内3H2O掺入量[(726±58)、(738±45)、(774±44)脉冲数/min]也显著低于DMEM对照组[(1148±70)脉冲数/min,P均<0.01]。(2)体内实验:ALI大鼠肺组织AQP1、AQP5mRNA表达(LPS2h组0.409±0.018、0.421±0.020,LPS4h组0.421±0.023、0.412±0.023,LPS6h组0.435±0.020、0.388±0.031,LPS8h组0.438±0.016、0.386±0.019)也显著低于对照组(0.794±0.015、0.787±0.022,P均<0.01)。结论AQP1、AQP5可能参与ALI/ARDS液体的异常转运,可能与肺水肿的发病机制有关。     

HIF-1αmRNA和GLUT1 mRNA在非小细胞肺癌组织中的表达及意义

目的探讨低氧诱导因子-1（HIF-1α）和葡萄糖转运蛋白子-1（GLUT1）的mRNA在非小细胞肺癌（NSCLC）组织中的表达以及与肺癌生物学特性的关系。方法采用PT—PCR、凝胶电泳半定量方法检测36例肺癌组织和12例癌旁组织中HIF-1αmRNA和GLUT1mRNA的表达，并分析它们在TNM分期中的表达情况。结果HIF-1αmRNA和GLUT1mRNA在癌症组织中的表达明显高于癌旁组织（P〈0.01），在有淋巴结转移组和有远端转移组中的表达均明显高于无淋巴结转移组（P〈0．05）和无远处转移组（P〈0．01）；HIF-1αmRNA与GLUT1mRNA的表达呈线性正相关（P〈0．01）。在T3、T4中的表达明显高于在T1、T2中的表达（P〈0．01或P〈0、05）。结论NSCLC组织中HIF-1αmRNA和GLUT1mRNA的表达与TNM分期、淋巴结转移和远端转移有关；HIF-1αmRNA和GLUT1mRNA表达的增高可能是NSCLC恶化的基因标志。     

糖尿病大鼠VFN水平与GLUT1、GLUT4基因表达的相关性

目的探讨内脏脂肪素(VFN)与葡萄糖转运蛋白(GLUT)1、GLUT4基因表达的相关性。方法健康雄性Wistar大鼠33只随机分为正常对照组(NC组),糖尿病模型组(DM组),高脂饮食组(HF组),分别为10只、13只、10只。检测3组大鼠体重、空腹血糖(FBG)和空腹血清胰岛素(FINS),用稳态模型评估法(HOMA)评价胰岛素抵抗(IR)指数,采尾血以酶联免疫吸附试验(ELISA)测定VFN的含量。处死,分离附睾、肠系膜、折返腹膜脂肪及肾脏脂肪垫,称重,计为内脏脂肪重量,利用RT-PCR法分别检测3组大鼠内脏脂肪中VFN的表达。RT-PCR方法检测各组大鼠心肌组织GLUT1、GLUT4的基因表达水平。结果成功建立大鼠糖尿病模型。DM组VFN表达明显增加,GLUT1、GLUT4的相对表达水平明显降低(P0.05)。结论糖尿病大鼠体内VFN水平可影响GLUT1、GLUT4 mRNA的表达,且具有负相关性。     

大承气汤对急性坏死性胰腺炎大鼠胰腺水通道蛋白1的影响

目的 观察急性坏死性胰腺炎( ANP)大鼠胰腺组织水通道蛋白1(AQP1)的表达以及大承气汤对其的影响.方法 160只雄性SD大鼠按随机数字法分为对照组、ANP组、地塞米松组、乙酰唑胺组及大承气汤组,每组32只.采用胆胰管逆行注射5％牛磺胆酸钠方法制备ANP模型.地塞米松组于造模后即刻静脉给予地塞米松4 mg/kg体重;乙酰唑胺组于造模前2h用含乙酰唑胺的生理盐水1ml灌胃;大承气汤组于造模前48、24、2h分别用大承气汤2ml/次灌胃;对照组仅开腹触摸胰腺数次后关腹.制模后3、6、12、18 h分批处死8只大鼠.记录腹水量;检测血清淀粉酶;胰腺组织病理检查及电镜观察;伊文思兰(EB)血管外渗法检测毛细血管通透性;实时PCR和蛋白质印迹法检测AQP1 mRNA和蛋白表达.结果 ANP组血清淀粉酶水平显著升高,胰腺损伤明显;地塞米松组和大承气汤组淀粉酶水平较ANP组降低,胰腺损伤减轻;乙酰唑胺组淀粉酶水平高于ANP组,胰腺病理损伤较ANP组加重.造模后6h,对照组、ANP组、地塞米松组、乙酰唑胺组、大承气汤组胰腺组织EB含量分别为(13.44±2.56)、( 126.35±14.80)、(86.31±14.46)、(108.99±15.07)、(78.29±16.85) mg/L;AQP1 mRNA表达量为(170.07±22.48)％、(83.93±8.98)％、(117.09±10.70)％、(69.00±8.98)％、(112.82±11.79)％;AQP1蛋白表达量为0.23±0.06、0.10±0.02、0.32±0.03、0.13±0.02、0.45±0.04.ANP组的EB量显著高于对照组,而AQP1mRNA及蛋白的表达显著低于对照组(P值均＜0.05);地塞米松组及大承气汤组EB含量显著低于ANP组,而AQP1 mRNA及蛋白的表达显著高于ANP组(P值均＜0.05).结论 AQP1在ANP大鼠胰腺组织毛细血管渗漏的发生中起重要作用.大承气汤通过调控AQP1的表达可减轻ANP大鼠的胰腺损伤.     

水通道蛋白1在去窦弓神经大鼠主动脉中的表达及意义

目的探讨水通道蛋白1（AQP1）蛋白在去窦弓神经（SAD）大鼠主动脉中的表达变化及其意义。方法选用SD大鼠制作SAD大鼠模型,取主动脉观察其变化,并通过RT-PCR及Western blot技术检测其中AQP1 mRNA及蛋白的表达变化。结果与假手术组比较,SAD组主动脉的单位长度质量及内壁厚度均增加（P均〈0.05）;主动脉中AQP1 mRNA及蛋白水平减少（P〈0.05或〈0.01）;主动脉AQP1 mRNA水平与主动脉的单位长度质量及内壁厚度呈负相关（P均〈0.01）。结论 AQP1在SAD大鼠主动脉平滑肌细胞增生的过程中可能起着重要作用。     

大鼠肾移植急性排斥模型中移植肾水通道蛋白-2 mRNA表达的变化

目的 研究水通道蛋白-2(AQP2)mRNA在大鼠肾移植急性排斥模型中移植肾表达的变化及意义.方法 正常大鼠肾脏为正常对照组;Wistar大鼠作为供、受者建立同系基因对照组;SD大鼠为供者、Wistar大鼠为受者建立同种异体移植急性排斥组;SD大鼠为供者、Wistar大鼠为受者,术后给予免疫抑制剂环孢素(CsA)建立同种异体移植免疫抑制组;均为雄性大鼠.RT-PCR半定量检测移植肾AQP2 mRNA在各组中表达的变化.结果 同种异体移植急性排斥组移植肾AQP2 mRNA表达下调,与正常对照组比较有统计学差异(P＜0.05),与同系基因对照组和同种异体移植免疫抑制组比较有统计学差异(P＜0.05).同系基因对照组和同种异体移植免疫抑制组中移植肾AQP2 mRNA表达的变化与正常对照组比较无统计学差异(P＞0.05).结论 AQP2 mRNA的表达在急性排斥时表达下调,可以减少对水的重吸收,同时可以减少集合系统能量消耗,有利于移植肾功能的恢复.     

脾胃湿热证与水通道蛋白4基因表达的关系

目的:从水液代谢来研究脾胃湿热证与水通道蛋白4(AQP4)基因的关系.方法:慢性浅表性胃炎患者25例,其中脾胃湿热证15例,脾气虚证10例,另10例健康志愿者为对照.荧光定量PCR法检测AQP4在各组胃粘膜组织中的mRNA表达量.结果:脾胃湿热证组胃粘膜组织中AQP4 mRNA表达量为(10.00±8.83)×106 cps/ml,显著高于脾气虚证组(1.28±2.93)×106 cps/ml(P＜0.01);而脾气虚证组则明显低于对照组[(2.99±3.33)×106 cps/ml,P<0.05].结论:AQP4基因表达的异常可能是脾胃湿热证的发生机制之一.     

洛哌丁胺对腹泻模型大鼠结肠水通道蛋白4表达的影响

目的检测腹泻模型大鼠在给予洛哌丁胺处理后结肠黏膜水通道蛋白4（AQP4）mRNA表达的变化,探讨洛哌丁胺在结肠水代谢巾的分子作用机制。方法采用逆转录聚合酶链反应（RT—PCR）分别对洛哌丁胺治疗组、模型对照组及正常对照组大鼠升、降结肠黏膜细胞AQP4mRNA表达进行半定量分析。结果①洛哌丁胺治疗组升、降结肠AQP4mRNA表达量均高于模型对照组及正常对照组（P〈0．01）。②模型对照组升结肠AQP4mRNA表达量较正常对照组大鼠减少（P〈0．05）,而降结肠表达差异无统计学意义（P〉0．05）。结论洛哌丁胺可以在转录水平上调腹泻大鼠结肠黏膜AQP4表达,使结肠对肠腔内水分的吸收增加。     

Cytochrome 1A1 and 1B1 gene diversity in the Zanzibar islands

Amodiaquine (AQ) is a 4‐aminoquinoline widely used in the treatment of malaria as part of the artemisinin combination therapy (ACT). AQ is metabolised towards its main metabolite desethylamodiaquine mainly by cytochrome P450 2C8 (CYP2C8). CYP1A1 and CYP1B1 play a minor role in the metabolism but they seem to be significantly involved in the formation of the short‐lived quinine‐imine. To complete the genetic variation picture of the main genes involved in AQ metabolism in the Zanzibar population, previously characterised for CYP2C8, we analysed in this study CYP1A1 and CYP1B1 main genetic polymorphisms. The results obtained show a low frequency of the CYP1A1*2B/C allele (2.4%) and a high frequency of CYP1B1*6 (approximately 42%) followed by CYP1B1*2 (approximately 27%) in Zanzibar islands. Genotype data for CYP1A1 and CYP1B1 show a low incidence of fast metabolisers, revealing a relatively safe genetic background in Zanzibar’s population regarding the appearance of adverse effects.     

CYP1A1 , GSTM1 , GSTT1 and NQO1 polymorphisms and colorectal adenomas in Japanese men

AIM: To investigate the role of functional genetic poly-morphisms of metabolic enzymes of tobacco carcinogens in the development of colorectal adenomas. METHODS: The study subjects were 455 patients with colorectal adenomas and 1052 controls with no polyps who underwent total colonoscopy in a preretirement health examination at two Self Defense Forces hospitals. The genetic polymorphisms studied wereCYP1A1*2A (rs 4646903), CYP1A1*2C (rs 1048943), GSTM1 (null or non-null genotype), GSTT1 (null or non-null genotype) and NQO1 C609T (rs 1800566). Genotypes were determined by the polymerase chain reaction (PCR)-restriction fragment length polymorphism or PCR method using genomic DNA extracted from the buffy coat. Cigarette smoking and other life-style factors were ascertained by a self-administered questionnaire. The associations of the polymorphisms with colorectal adenomas were examined by means of OR and 95%CI, which were derived from logistic regression analysis. Statistical adjustment was made for smoking, alcohol use, body mass index and other factors. The gene-gene interaction and effect modification of smoking were evaluated by the likelihood ratio test. RESULTS: None of the five polymorphisms showed a significant association with colorectal adenomas, nor was the combination of GSTM1 and GSTT1 . A borderline significant interaction was observed for the combination of CYP1A1*2C and NQO1 (P = 0.051). The OR associated with CYP1A1*2C was significantly lower than unity among individuals with the NQO1 609CC genotype. The adjusted OR for the combination of the CYP1A1*2C allele and NQO1 609CC genotype was 0.61 (95%CI: 0.42-0.91). Although the interaction was not statistically significant (P = 0.24), the OR for individuals carrying the CYP1A1*2C allele and GSTT1 null genotype decreased significantly compared with those who had neither CYP1A1*2C allele nor GSTT1 null genotype (adjusted OR: 0.69, 95%CI: 0.49-0.97). Smoking did not modify the associations of the individual polymorphisms with colorectal adenomas. There w     

Inhibition of procarcinogen-bioactivating human CYP1A1, CYP1A2 and CYP1B1 enzymes by melatonin

Abstract:  Administration of melatonin to rodents decreases the incidence of tumorigenesis initiated by benzo[ a ]pyrene or 7,12-dimethylbenz[ a ]anthracene, which requires bioactivation by cytochrome P450 enzymes, such as CYP1A1, CYP1A2 and CYP1B1, to produce carcinogenic metabolites. The present study tested the hypothesis that melatonin is a modulator of human CYP1 catalytic activity and gene expression. As a comparison, we also investigated the effect of melatonin on the catalytic activity of CYP2A6, which is also a procarcinogen-bioactivating enzyme. Melatonin (3–300 μ m ) decreased 7-ethoxyresorufin O -dealkylation catalyzed by human hepatic microsomes and recombinant CYP1A1, CYP1A2 and CYP1B1, whereas it did not affect coumarin 7-hydroxylation catalyzed by hepatic microsomes or recombinant CYP2A6. Melatonin inhibited CYP1 enzymes by mixed inhibition, with apparent K _i values (mean ± S.E.M.) of 59 ± 1 (CYP1A1), 12 ± 1 (CYP1A2), 14 ± 2 (CYP1B1) and 46 ± 8 μ m (hepatic microsomes). Additional experiments indicated that melatonin decreased benzo[ a ]pyrene hydroxylation catalyzed by hepatic microsomes and CYP1A2 but not by CYP1A1 or CYP1B1. Treatment of MCF-10A human mammary epithelial cells with melatonin (up to 300 μ m ) did not affect basal or benzo[ a ]pyrene-inducible CYP1A1 or CYP1B1 gene expression. Consistent with this finding, melatonin did not influence reporter activity in aryl hydrocarbon receptor-dependent pGudluc6.1-transfected MCF-10A cells treated with or without benzo[ a ]pyrene, as assessed in an in vitro cell-based luciferase reporter gene assay. Overall, melatonin is an in vitro inhibitor of human CYP1 catalytic activity, and it may be useful to develop potent analogues of melatonin as potential cancer chemopreventive agents that block CYP1-mediated chemical carcinogenesis.     

The target of ezetimibe is Niemann-Pick C1-Like 1 (NPC1L1)

Ezetimibe is a potent inhibitor of cholesterol absorption that has been approved for the treatment of hypercholesterolemia, but its molecular target has been elusive. Using a genetic approach, we recently identified Niemann-Pick C1-Like 1 (NPC1L1) as a critical mediator of cholesterol absorption and an essential component of the ezetimibe-sensitive pathway. To determine whether NPC1L1 is the direct molecular target of ezetimibe, we have developed a binding assay and shown that labeled ezetimibe glucuronide binds specifically to a single site in brush border membranes and to human embryonic kidney 293 cells expressing NPC1L1. Moreover, the binding affinities of ezetimibe and several key analogs to recombinant NPC1L1 are virtually identical to those observed for native enterocyte membranes. KD values of ezetimibe glucuronide for mouse, rat, rhesus monkey, and human NPC1L1 are 12,000, 540, 40, and 220 nM, respectively. Last, ezetimibe no longer binds to membranes from NPC1L1 knockout mice. These results unequivocally establish NPC1L1 as the direct target of ezetimibe and should facilitate efforts to identify the molecular mechanism of cholesterol transport.     

Interleukin 1 (IL 1)

Interleukin 1 is an essential factor of macrophage dependent T cell activation and has a large quantity of other biological activities. This paper gives a review of present knowledge of Interleukin 1. In addition to biochemical properties, the IL 1 production and IL 1 activities, methods for determining of IL 1 and inhibitory factors of IL 1 induced T cell proliferation are described.     

Pandemic H1N1 influenza

The 2009 H1N1 influenza A virus that has targeted not only those with chronic medical illness, the very young and old, but also a large segment of the patient population that has previously been afforded relative protection - those who are young, generally healthy, and immune naive. The illness is mild in most, but results in hospitalization and severe ARDS in an important minority. Among those who become critically ill, 20-40% will die, predominantly of severe hypoxic respiratory failure. However, and potentially in part due to the young age of those affected, intensive care with aggressive oxygenation support will allow most people to recover. The volume of patients infected and with critical illness placed substantial strain on the capacity of the health care system and critical care most specifically. Despite this, the 2009 pandemic has engaged our specialty and highlighted its importance like no other. Thus far, the national and global critical care response has been brisk, collaborative and helpful - not only for this pandemic, but for subsequent challenges in years ahead.