氯沙坦抑制高胆固醇血症大鼠动脉核转录因子活化和细胞间粘附分子表达

目的 :观察氯沙坦 (Los)对高胆固醇血症大鼠血管壁核转录因子 (NFkB)活化和细胞间粘附分子 (ICAM 1)表达的影响。方法 :3 0只雄性成年SD大鼠随机分为正常对照组 (NC组 ) ,高脂组 (HL组 )和Los组。 10周后 ,酶法测血清总胆固醇 ,硝酸还原酶法测胸主动脉组织中一氧化氮 (NO)、硫代巴比妥酸法测丙二醛 (MDA)含量及用黄嘌呤氧化酶法测超氧化物歧化酶 (SOD)活性 ;用免疫组化法测主动脉壁NFkB活化程度及ICAM 1蛋白表达水平 ;HE染色高倍视野下计数浸润于内膜的单核细胞数。结果 :Los组胸主动脉NO含量和SOD活力显著高于HL组 ,而MDA水平低于HL组 ;血管壁NFkB活化和ICAM 1表达水平及血管内膜中单核细胞数明显低于HL组 ;这 2组血清胆固醇浓度显著高于NC组 ,但它们之间无差异。结论 :Los能缓解自由基损伤 ,抑制高胆固醇血症大鼠主动脉NFkB活化和ICAM 1表达 ,它可通过该机制防止动脉粥样硬化形成和发展     

氯沙坦对家兔早期动脉粥样硬化血管MCP-1表达及胆固醇含量的影响

目的 :氯沙坦对加速性家兔早期动脉粥样硬化 （AS）血管增生及 MCP- 1蛋白表达的影响。方法 :采用组织学、免疫组织化学和生物化学方法评价加速性家兔 AS早期血管组织学、增殖细胞核抗原 （PCNA）及单核白细胞趋化蛋白 - 1（MCP- 1）蛋白表达和组织胆固醇含量的改变。结果 :AS组血管内膜与中膜厚度比值、主动脉组织胆固醇含量、内膜 PCNA阳性细胞数及 MCP- 1蛋白表达平均光密度积分值均显著增加 ,氯沙坦干预组内膜与中膜厚度比值较 AS组显著下降 ,PCNA阳性细胞数明显减少 ,血管壁 MCP- 1蛋白表达水平明显下降 （均 P<0 .0 1）。氯沙坦干预组主动脉组织胆固醇含量明显低于 AS组 [1.47± 0 .0 3mg/ g（wet）与 1.35± 0 .0 2 mg/ g（wet） ,P<0 .0 5 ]。结论 :氯沙坦干预可抑制 MCP- 1蛋白表达 ,减轻加速性 AS病变血管内膜增生 ,减少胆固醇在血管壁沉积 ,从而可能在防治AS过程中起重要作用。     

氯沙坦对血管平滑肌细胞内单核细胞趋化因子-1表达的影响

为了探讨血管紧张素Ⅱ受体拮抗剂氯沙坦对血管平滑肌细胞内单核细胞趋化蛋白 1表达的影响 ,以培养幼兔主动脉平滑肌细胞为研究对象 ,分别给予不同浓度血管紧张素Ⅱ和 或血管紧张素Ⅱ受体拮抗剂氯沙坦 ,采用免疫组织化学、原位杂交与酶联免疫吸附技术检测不同处理组平滑肌细胞内单核细胞趋化蛋白 1蛋白及其mR NA表达和平滑肌细胞培养介质中单核细胞趋化蛋白 1含量的变化。结果发现 ,10 - 6 ～ 10 - 1 0 mol L血管紧张素Ⅱ呈剂量依赖性地增加平滑肌细胞内单核细胞趋化蛋白 1蛋白及其mRNA表达水平 ,增高培养介质中单核细胞趋化蛋白 1蛋白含量 (P均 <0 .0 0 1)。 10 - 5 ～ 10 - 7mol L氯沙坦预处理使血管紧张素Ⅱ刺激的平滑肌细胞内单核细胞趋化蛋白 1蛋白及其mRNA表达水平及培养介质中单核细胞趋化蛋白 1蛋白含量明显减低 (P均 <0 .0 0 1)。以上结果提示 ,氯沙坦可拮抗血管紧张素Ⅱ所致的平滑肌细胞内单核细胞趋化蛋白 1的表达与分泌 ,这可能有助于减轻或防治某些病理状态下血管平滑肌细胞的增殖与迁移。     

NF-κB和Jak-STAT信号途径参与血管紧张素Ⅱ介导的大鼠主动脉平滑肌细胞增殖效应

目的进一步探讨JakSTAT信号途径在血管紧张素Ⅱ介导的主动脉平滑肌细胞增殖效应的作用。方法培养大鼠主动脉平滑肌细胞并行免疫组织化学鉴定,用血管紧张素Ⅱ以不同时间梯度刺激传代培养的大鼠主动脉平滑肌细胞,采用BrdU法测定细胞增殖,裂解细胞后提取细胞总蛋白,分别经免疫共沉淀、Western印迹和细胞免疫荧光化学等方法分析胞内NFκB和Jak/STAT信号分子激活及表达的情况,行不同组间并与阴性对照组做对比。结果在6～30h的平滑肌细胞增殖实验中,6、12h时间段增值最明显;其中12h时AngⅡ组490nm处吸光度值(0.590±0.029)显著高于AG490 AngⅡ组(0.381±0.019),PDTC AngⅡ组(0.481±0.024),氯沙坦 AngⅡ组(0.519±0.026)和SerumFree组(0.30±0.02),P<0.01。Western印迹显示AngⅡ组中NFκB及磷酸化的Jak2、STAT1分别在5、15、60min表达明显达高峰;免疫荧光则表明NFκB及STAT1分子随时间梯度可逆的由胞浆转至胞核,在AngⅡ刺激15min组中STAT1表达水平比对照组(P<0.01)及刺激60min高(P<0.05)。结论AngⅡ能导致VSMC增殖,同时NFκB和磷酸化的Jak2,STAT1表达增加并随刺激时间梯度改变。因此可说明在NFκB和Jak/STAT信号通路激活参与了血管紧张素Ⅱ导致的主动脉平滑肌细胞增殖作用。     

实验性高脂血症兔MCP-1与CD11b的表达及氯沙坦的干预作用

目的观察氯沙坦对高胆固醇血症兔主动脉组织单核细胞趋动蛋白1（MCP-1）基因表达水平和静脉血单核细胞表面黏附分子CD11b活性（CD11b）的影响。方法40只日本长耳白兔随机分为4组,每组10只。正常对照组喂以普通饲料;高胆固醇组喂以高胆固醇饲料;氯沙坦高、低剂量组在喂高胆固醇饲料同时,分别以氯沙坦粉剂25 mg/（kg.d）和10 mg/（kg.d）溶于温水中灌胃,1次/d。观察12周。实验结束时,主动脉组织经HE染色行病理学检查。用流式细胞技术检测CD11b。用RT-PCR方法检测主动脉组织MCP-1的基因表达。结果高胆固醇组血管内膜下斑块形成明显,而氯沙坦高、低剂量组斑块明显减少;高胆固醇组较对照组MCP-1 mRNA表达及CD11b活性显著增高（P<0.01）;氯沙坦高、低剂量组MCP-1 mRNA表达及CD11b活性较高胆固醇组显著降低（P<0.05）。结论氯沙坦抑制组织MCP-1 mRNA的表达及降低CD11b活性,降低单核细胞内膜下浸润,其作用机制可能是氯沙坦从受体水平阻断肾素-血管紧张素-醛固酮系统。     

植物血凝素样受体在兔动脉粥样硬化斑块中的表达及氯沙坦对其影响

目的 :观察植物血凝素样受体 (Lox 1)在兔动脉粥样硬化 (AS)组织中的表达和氯沙坦对Lox 1及斑块的影响。方法 :将 2 2只雄性新西兰白兔按体重随机分为对照组、高胆固醇组和氯沙坦组。检测指标 :①于 0、6、10周取血测定血浆血管紧张素Ⅱ (AngⅡ )、血脂水平 ;②测量主动脉AS斑块面积和内膜、中膜厚度 ;③免疫组化和逆转录 聚合酶链反应 (RT PCR)法检测动脉Lox 1蛋白及mRNA的表达。结果 :高脂饮食明显增加血脂及AngⅡ的水平 ,动脉内膜增生程度、斑块面积也随之增加 ,Lox 1(mRNA、蛋白 )表达较对照者更明显 (均 P <0 .0 1)。应用氯沙坦后血脂较高胆固醇组无明显变化 (P >0 .0 5 ) ,AngⅡ水平却明显升高 (P <0 .0 1) ,而动脉内膜增生、Lox 1表达不及高胆固醇组明显 (P <0 .0 1)。结论 :Lox 1与AS的发生、发展有着紧密的联系 ,高脂饮食促进Lox 1表达。氯沙坦可抑制Lox 1表达 ,具有抗AS的作用     

血管紧张素Ⅱ受体拮抗剂对自发性高血压大鼠的血管炎症抑制作用

目的观察血管紧张素Ⅱ受体拮抗剂(ARB)对自发性高血压大鼠血管炎性反应以及单核细胞趋化蛋白1(MCP-1)/趋化因子CC亚家族受体2(CCR2)途径的影响,探讨MCP-1/CCR2途径在自发性高血压血管炎性反应中的作用。方法自发性高血压大鼠(SHR)24只,随机分为高血压对照组(HC)、低剂量氯沙坦组(LL)、高剂量氯沙坦组(HL)和替米沙坦组(T组),正常血压大鼠(WKY)6只为正常对照组(NC)。每天1次灌胃给药:LL组氯沙坦5 mg/kg,HL组氯沙坦30 mg/kg,T组替米沙坦30 mg/kg,HC组及NC组等量蒸馏水。4周后处死动物,免疫组化检测大鼠胸主动脉壁巨噬细胞浸润(ED-1标记),反转录-聚合酶链反应(RT-PCR)检测心、肾、胸主动脉、循环血单个核细胞MCP-1、CCR2的表达,对外周血单个核细胞进行MCP-1的趋化功能实验,酶联免疫吸附法(ELISA)观察大鼠血清MCP-1水平。结果SHR组与WKY组相比,胸主动脉壁ED-1阳性细胞明显增多(P<0.01),心、肾、胸主动脉、循环血单个核细胞MCP-1 mRNA、CCR2 mRNA明显升高(均为P<0.01),外周血单个核细胞对MCP-1的趋化性明显升高(P<0.01),血清MCP-1水平升高(P<0.01)。不同剂量氯沙坦及替米沙坦均能有效抑制各组织和循环中MCP-1及CCR2的表达、外周血单个核细胞对MCP-1的趋化性以及胸主动脉壁ED-1阳性细胞数(与HC组相比,LL组、HL组及T组均为P<0.01),上述作用不依赖ARB的降压作用。结论ARB通过调节MCP-1/CCR2途径抑制血管炎性反应,此作用独立于其降压作用。     

氯沙坦对球囊成形术后兔血管内膜增生及核因子κB活性的影响

为研究氯沙坦对球囊成形术后血管内膜增生及核因子κB活性的影响 ,采用内皮剥脱后高脂饮食 (含 1.5 %胆固醇 )喂养制作兔腹主动脉粥样硬化模型 ,然后对狭窄部位行球囊成形术。术后氯沙坦组给予氯沙坦 10mg/(kg·d)口服 ,对照组只喂生理盐水 ,4周后取腹主动脉行组织形态学观察及用免疫组织化学方法分析核因子κB、α 平滑肌肌动蛋白、巨噬细胞、细胞间粘附分子 1的表达。结果发现 ,与对照组相比 ,氯沙坦组内膜厚度 /中膜厚度比值、内膜面积 /中膜面积比值均显著减少 (P <0 .0 1)。氯沙坦组新生内膜中巨噬细胞、平滑肌细胞数均较对照组显著减少 (P <0 .0 1和P <0 .0 5 )。核因子κB及其靶基因产物细胞间粘附分子 1水平亦因氯沙坦的干预而明显减少(P <0 .0 1和P <0 .0 5 )。结果提示 ,氯沙坦明显抑制球囊成形术后内膜增生 ,其机制可能为阻断血管紧张素Ⅱ受体 ,进而抑制核因子κB ,从而抑制平滑肌细胞迁移、增殖和新生内膜形成。     

高浓度葡萄糖条件下血管紧张素Ⅱ对内皮细胞核因子κB和单核细胞趋化蛋白1的影响

观察高糖环境下血管紧张素Ⅱ对内皮细胞核因子κB活性、单核细胞趋化蛋白1表达及氯沙坦的干预作用。应用激光共聚焦显微镜观察核因子κB活性变化，逆转录．聚合酶链反应测定内皮细胞单核细胞趋化蛋白1mRNA表达，酶联免疫吸附法检测培养基中单核细胞趋化蛋白1含量变化。结果发现，高糖(25mmol/L)、10^-7mol/L血管紧张素Ⅱ均能显著刺激内皮细胞核因子κB活化、单核细胞趋化蛋白1mRNA表达，培养基中单核细胞趋化蛋白1含量及其对单核细胞趋化活性亦显著增强，高糖环境显著增加血管紧张素Ⅱ诱导的核因子κB活化、单核细胞趋化蛋白1表达，氯沙坦有显著抑制作用。提示高糖促进血管紧张素Ⅱ对内皮细胞的刺激作用，氯沙坦通过抑制内皮细胞核因子κB活化和单核细胞趋化蛋白1的表达．对动脉粥样硬化发展起到防治作用．     

阿托伐他汀对自发性高血压大鼠血压和心肌血管紧张素Ⅱ1型和2型受体表达的影响

目的　观察阿托伐他汀对自发性高血压大鼠血压和心肌血管紧张素Ⅱ受体 1和血管紧张素Ⅱ受体 2的调节作用。方法　采用免疫组织化学染色法检测心肌血管紧张素Ⅱ受体 1和血管紧张素Ⅱ受体 2蛋白表达 ,原位杂交法测定心肌血管紧张素Ⅱ受体 1和血管紧张素Ⅱ受体 2mRNA表达水平。于给药前和给药后每两周测量大鼠尾动脉收缩压 ,并测定血清总胆固醇、甘油三酯、高密度脂蛋白胆固醇及低密度脂蛋白胆固醇水平。结果　实验前自发性高血压大鼠各组收缩压均显著高于Wistar kyoto大鼠组 (P <0 .0 1)。给药后第 4周和第 6周 ,5 0mg阿托伐他汀组收缩压明显下降 (P <0 .0 1) ,总胆固醇、甘油三酯及低密度脂蛋白胆固醇水平明显降低 (P <0 .0 5 ,P <0 .0 1) ;自发性高血压大鼠对照组心肌血管紧张素Ⅱ受体 1和血管紧张素Ⅱ受体 2蛋白阳性表达及其mRNA表达均明显高于Wistar kyoto大鼠组 (P <0 .0 1) ,6周后 ,5 0mg阿托伐他汀组血管紧张素Ⅱ受体 1蛋白和其mRNA表达明显降低 (P <0 .0 1) ,而血管紧张素Ⅱ受体 2蛋白和其mRNA表达明显高于自发性高血压大鼠对照组 (P <0 .0 1)。结论　阿托伐他汀能降低自发性高血压大鼠的血压 ,并对心肌血管紧张素Ⅱ受体有双重调节作用 ,即使血管紧张素Ⅱ受体 1下调、血管紧张素Ⅱ受体 2上调     

The immunoneuroendocrine role of melatonin

Abstract: A tight, physiological link between the pineal gland and the immune system is emerging from a series of experimental studies. This link might reflect the evolutionary connection between self-recognition and reproduction. Pinealectomy or other experimental methods which inhibit melatonin synthesis and secretion induce a state of immunodepression which is counteracted by melatonin. In general, melatonin seems to have an immunoenhancing effect that is particularly apparent in immunodepressive states. The negative effect of acute stress or immunosuppressive pharmacological treatments on various immune parameters are counteracted by melatonin. It seems important to note that one of the main targets of melatonin is the thymus, i.e., the central organ of the immune system. The clinical use of melatonin as an immunotherapeutic agent seems promising in primary and secondary immunodeficiencies as well as in cancer immunotherapy. The immunoenhancing action of melatonin seems to be mediated by T-helper cell-derived opioid peptides as well as by lymphokines and, perhaps, by pituitary hormones. Melatonin-induced-immuno-opioids (MHO) and lymphokines imply the presence of specific binding sites or melatonin receptors on cells of the immune system. On the other hand, lymphokines such as -γ-interferon and interleukin-2 as well as thymic hormones can modulate the synthesis of melatonin in the pineal gland. The pineal gland might thus be viewed as the crux of a sophisticated immunoneuroendocrine network which functions as an unconscious, diffuse sensory organ.     

Response of rat pineal melatonin to calcium, magnesium, and lithium is circadian stage dependent

Abstract: Herein we documented the response of pineal melatonin production to electrolytes known to be effective on pineal function in view of a possible circadian stage dependence. We studied the release of melatonin by perifused rat pineal glands at 2 different circadian stages corresponding to the middle of the light and dark periods, i.e., respectively, 7 and 19 HALO (Hours After Light Onset, L:D = 12:12). The initial efflux rates were, as expected, much higher in the perifusates of glands removed from rats sacrificed during the dark phase than of those removed during the light phase. After 3 hr of perifusion, melatonin release reached similar levels which were found constant up to the 8th hr of perifusion, whatever the circadian stage. Perifusion of the glands with physiological concentrations for the rat of calcium (5.2 mmol/1) and magnesium (1.34 mmol/1) resulted in a stimulatory effect on the pineal glands removed from rats sacrificed in the middle of the dark period (19 HALO), whereas no effects were observed on the pineal glands removed from rats sacrificed during the light (7 HALO). Lithium (0.28 and 0.55 mmol/1) was ineffective on melatonin release in pineal glands removed 7 and 19 HALO. Our results show differences in the initial efflux rates of melatonin and in the response of perifused pineal glands to calcium and magnesium according to the circadian stage.     

Changes in connexin43, the gap junction protein of astrocytes, during development of the rat pineal gland

Abstract: The abundance of gap junctions between rat pineal astrocytes formed by connexin43 (Cx43) was studied during development. Levels and distribution of Cx43 were measured by immunoblotting and indirect immunofluorescence, respectively. The amount of Cx43 in cells located within the gland was low until about the 7th postnatal day and increased to adult values between the 14th and 21st days postpartum. Although astrocytes, recognized by their vimentin immunoreactivity, were scarce before birth, they were abundant by the 7th postnatal day suggesting that the low levels of Cx43 found at this age corresponded to a low expression of this protein. Localization of the immunoreactivity to Cx43 and vimentin showed a close correlation, indicating that mature or immature pineal astrocytes form gap junctions made of Cx43. Since Cx43 levels attained their adult values at about the time the innervation and the functional state of the gland reached maturity (2–3 weeks after birth), it is proposed that astrocyte gap junctions are involved in the function of the adult rat pineal gland.     

Conservative management of perforated duodenal diverticulum： A case report and review of the literature

Duodenal diverticula are a relatively common condition. They are asymptomatic, unless they become complicated, with perforation being the rarest but most severe complication. Surgical treatment is the most frequently performed approach. We report the case of a patient with a perforated duodenal diverticulum, which was diagnosed early and treated conservatively with antibiotics and percutaneous drainage of secondary retroperitoneal abscesses. We suggest this method could be an acceptable option for the management of similar cases, provided that the patient is in good general condition and without septic signs.     

Presence of immunoreactive growth hormone and prolactin in the ovine pineal gland

Abstract: The use of antisera raised against bovine growth hormone (GH) and ovine prolactin (PRL) enabled the detection of related immunoreactive (ir) sequences of proteins in ovine pineal tissue. The isolation of PRL-like ir-material was accomplished using a 0.25 M ammonium sulphate (pH 5.5) extraction followed by ethanol precipitation, whereas the resulting 2.0 M ammonium sulphate (pH 7.0) precipitate contained a GH-like immunoreactivity. Gel chromatography of the GH-like immunoreactivity (Sephadex G-100) indicated the presence of several GH-like fragments ranging in the M_r range of 7,000 to 55,000. Analyses of the PRL-like ir-material found in pineal tissue on HPLC using a TSK 545-DEAE column led to the resolution into a single peak of immunoreactivity. A single peak of activity was also observed following chromatofocusing and hydrophobic interaction chromatography of the ir-peak from the TSK 545-DEAE column. The PRL-like ir-material inhibited the binding of [¹²⁵I]ovine PRL-S14 to anti-ovine PRL antibodies without showing an affinity for binding to anti-rat PRL or anti-bovine GH antibodies. Scatchard analysis of the binding of pineal PRL-like ir-material and pituitary ovine PRL-S14 to liver membranes from day-20 pregnant rats revealed similar affinity constants (K_a of 4.7 ± 0.2 × 10⁹ M^-1). In addition, the replication of Nb 2 Node rat lymphoma cells was stimulated by pineal PRL-like ir-material, an effect known to be specific for lactogenic hormones. The pineal PRL-like immunoreactivity appeared on sodium dodecyl sulfate polyacrylamide gels as a single major band of M_r 24,000. The functional status of PRL-and GH-like ir-material in the ovine pineal remains to be determined, but evidence is presented that the overall protein synthesis rate of the rat pineal responded to circulating concentrations of PRL.     

Microbiology of human immunodeficiency virus anorectal disease

PURPOSE: Individuals who are seropositive for the human immunodeficiency virus are at high risk for opportunistic infection and anorectal disorders. Little prospective information is available regarding anorectal pathogens in these patients. METHODS: One hundred sixty-three HIV-seropositive patients presented to the colorectal clinic between 1989 and 1992. Forty-seven (29 percent) patients were thought to have an infectious process and were prospectively studied using a standardized multiculture protocol. RESULTS: Mean age was 33 (range, 19–59) years. All were male; high-risk behavior accounted for 87 percent of HIV transmissions. Presenting complaints included anorectal pain (79 percent), pus per anum (28 percent), and blood per anum (26 percent). Examination revealed perianal tenderness (60 percent), condyloma (38 percent), perianal ulcers (38 percent), and anal fissures (34 percent). Sixty-six sets of cultures were performed; 28 patients had one set, 15 had two sets, and 4 had three sets. Thirty-two of these 47 patients (68 percent) had positive cultures including herpes (50 percent), cytomegalovirus (25 percent),Neisseria gonorrhoeae (16 percent), chlamydia (16 percent), acidfast bacilli (2 percent), and others (9 percent). Six of 32 patients with positive cultures had more than one organism cultured. Sixteen (50 percent) patients with positive cultures were treated medically, 8 (25 percent) were treated surgically and 8 (25 percent) were treated with both modalities. Sixty-one procedures were performed on 17 patients for condylomata. Eighteen patients had 20 procedures for abscesses, 50 percent of whom had positive cultures for other than common bowel flora; all improved. Fourteen patients underwent 33 procedures for perianal fistulas.Mycobacterium fortuitum was cultured from one patient who required 13 procedures for abscesses and fistulas. Forty-five (96 percent) patients were followed for an average of 12.5 months ±2.9 SEM (range, 1–94 months). Symptoms were improved or resolved in 22 of 32 (69 percent) patients with positive cultures and in 11 of 13 (84 percent) with negative cultures. CONCLUSIONS: Specific pathogens may often be identified in human immunodeficiency virus-seropositive patients with anorectal disorders if aggressively sought. Although patients without specific pathogens identified may be expected to improve with planned empiric treatment, positive identification allows more directed therapy.