塞来昔布对急性肺损伤大鼠肺组织NF-κB和iNOS的影响

【摘要】目的探讨脂多糖（LPS）所致大鼠急性肺损伤时环氧化酶-2（COX-2）抑制剂塞来昔布对肺组织的核因子-κB（NF-κB）p65和诱导型一氧化氮合酶（iNOS）表达的影响。方法将60只大鼠随机分为对照组、LPS组、治疗组和塞来昔布组,每组15只。LPS组尾静脉注射LPS（5mg/kg）复制急性肺损伤模型;治疗组注射LPS复制急性肺损伤模型后30min用塞来昔布灌胃（20mg/kg）;塞来昔布组不造模,于相同时间点用相同剂量塞来昔布灌胃;对照组不造模、不给药,给等量的生理盐水;各组均于3h后放血处死动物。采用蛋白质印迹（Western blot）和逆转录-聚合酶链反应（RT-PCR）方法分别检测肺组织COX-2、NF-κB p65、iNOS蛋白及NF-κB p65、iNOS mRNA的表达。结果LPS组与对照组相比COX-2、NF-κB p65和iNOS蛋白及NF-κB p65、iNOS mRNA表达均显著升高（P<0.01）;治疗组NF-κB p65、iNOS mRNA和蛋白表达较LPS组明显降低（P<0.01）。结论选择性COX-2抑制剂塞来昔布对急性肺损伤大鼠有保护作用。     

阿嗪米特对豚鼠胆囊胆固醇结石的治疗作用

目的:研究阿嗪米特(azintamide)对豚鼠胆囊胆固醇结石的治疗作用。方法:用高胆固醇饮食诱发豚鼠胆囊胆固醇结石模型,进行胆囊结石计数;测定胆囊体积和胆囊胆汁量;测定血清总胆汁酸(TBA)、总胆红素(TBIL)、胆固醇(TCHO)、三酰甘油(TG)、高密度脂蛋白胆固醇(HDL-c)、低密度脂蛋白胆固醇(LDL-C)、碱性磷酸酶(ALP);测定胆汁TCHO,TBA和TBIL。观察阿嗪米特(20和80mg·kg~(-1))对胆囊胆固醇结石的治疗作用。为探讨阿嗪米特抗胆囊胆固醇结石的作用机制,对胆囊壁组织进行了黏液素(mucin)的免疫组化实验。结果:与模型组比较,阿嗪米特20和80mg·kg~(-1)组成石率明显下降;胆囊体积和胆囊胆汁量显著减少;血清TCHO及LDL-C明显降低(P＜0．05);胆汁TBA升高(P＜0．05),胆汁TCHO及TBIL降低(P＜0．05)。黏液素免疫组化实验中可观察到,模型组黏液素表达水平较高,而阿嗪米特组黏液素表达水平较低。已知利胆药熊去氧胆酸(UDCA)亦有显著效果。结论:阿嗪米特对高胆固醇饮食诱发的豚鼠胆囊胆固醇结石模型具有明显的治疗作用。     

痰热清注射液对AECOPD肺部感染合并T2DM疗效机制及巨噬细胞异质性途径干预作用探讨

目的：观察痰热清注射液对AECOPD（慢性阻塞性肺疾病急性加重期）合并T2DM（2型糖尿病）患者的疗效机制探讨其对肺泡巨噬细胞异质性途径的干预作用。方法：选取某院呼吸科2017年09月-2018年3月住院患者90例,随机分为对照组和观察组,治疗7 d后分别观察2组临床疗效,WBC计数、肺功能,并使用ELisa法检测2组患者外周血中的中的MSP（巨噬细胞刺激蛋白）、NF-κB（核因子κB）、iNOS（诱导型一氧化氮合酶）、COX-2（环氧化酶2）的含量。结果：2组患者治疗前肺部感染症状体征、白细胞计数、肺功能、MSP、NF-κB,iNOS、COX-2表达无明显差异,且MSP与NF-κB,iNOS、COX-2表达呈正相关（r=0.992、r=0.992、r=0.983）,证明NF-κB,iNOS、COX-2的表达与MSP存在显著相关性;治疗后症状体征积分比较,在喘息、咳嗽、咳痰、哮鸣音方面观察组优于对照组（P<0.05）,白细胞计数观察组明显优于对照组（P<0.01）,差异具有显著性。MSP、NF-κB,iNOS、COX-2观察组明显优于对照组（P<0.01）,差异具有显著性。结论：肺泡异质途径在肺部感染中发挥了一定作用,痰热清注射液能够有效治疗AECOPD合并T2DM患者肺部感染,干预肺泡异质途径,从而提高临床疗效。     

自由基对豚鼠胆色素结石形成的影响

复制林可霉素致豚鼠胆囊胆色素结石的模型（Ｂ组）;用维生素Ｅ（ＶＥ）抗氧化预防结石形成（Ｃ组）。在结石形成的早期,测定其胆囊胆汁中的丙二醛（ＭＤＡ）、粘蛋白（ＭＰ）、β－葡萄糖醛酸酶（β－Ｇ）和总胆红素（ＴＢ）的含量。结果表明：Ｂ、Ｃ两组胆囊胆汁中ＭＤＡ、ＭＰ、β－Ｇ和ＴＢ的含量比正常对照组（Ａ组）均显著增高。Ｂ、Ｃ两组间各成分无差异。Ｂ、Ｃ组ＭＤＡ含量的对数与ＭＰ或β－Ｇ含量的对数分别呈显著正相关。说明炎症等病理过程产生的自由基通过对胆道粘膜上皮的损伤,扰乱其功能,在林可霉素致豚鼠胆色素结石的过程中起一定的作用。     

特发性肺纤维化患者肺组织环氧合酶-2表达的研究

目的探讨特发性肺纤维化(IPF)患者肺组织环氧合酶-2(COX-2)表达情况。方法提取20例特发性肺纤维化患者肺组织及正常组织RNA,应用逆转录聚合酶链反应(RT-PCR)技术检测COX-2mRNA的表达;采用放射免疫法检测患者支气管肺泡灌洗液(BALF)及血清中前列腺素E2(PGE2)水平。结果 20例特发性肺纤维化患者肺组织COX-2mRNA表达的相对量(%)为(143.10±7.16),高于正常肺组织COX-2mRNA表达的相对量(32.90±1.65),统计学比较有差异有统计学意义(P<0.05);BALF中PGE2浓度(ng/L)为(639.50±96.27),高于血清中PGE2浓度(257.36±41.40),差异有统计学意义(P<0.05)。结论特发性肺纤维化患者肺组织存在COX-2的高表达。     

槲寄生凝集素抑制人大肠癌HCT116细胞环氧化酶-2表达的研究

目的：探讨槲寄生凝集素对人大肠癌HCT116细胞环氧化酶（COX-2）表达的影响。方法：采用RT-PCR、Western blotting和ELISA等方法检测不同浓度（0.1～1mg/L）槲寄生凝集素对细胞COX-2及其产物前列腺素E2（PGE2）表达的影响。结果：槲寄生凝集素可抑制HCT116细胞的COX-2mRNA、蛋白表达及PGE2的水平,其抑制COX-2蛋白的表达与PGE2的水平相一致,只在大剂量（1mg/L）时对COX-1mRNA有抑制作用。结论：槲寄生凝集素具有抑制HCT116细胞的COX-2基因转录、蛋白表达和功能活性等作用,在一定浓度（0.1～1mg/L）和时间范围（3～24h）内,表现出时效与量效关系。     

呼吸道合胞病毒感染巨噬细胞诱导COX-2表达的相关研究

目的探讨呼吸道合胞病毒（respiratory syncytial virus,RSV）感染肺巨噬细胞RAW264．7细胞时,环氧化酶-2（cyclooxygenase-2,COX-2）的表达及其相关炎症介质PGE2（Prostaglandin E2）的水平变化,初步阐明COX-2在RSV感染所致炎性反应中的作用,为研究RSV感染时的炎症产生机理,临床治疗RSV感染所致的炎症反应等提供一定的理论依据。方法RSV感染体外培养的RAW264．7巨噬细胞,分别于不同感染时间点（4、8、16、24h）收集细胞和细胞培养上清。用半定量逆转录聚合酶链反应（RT．PCR）和Western-blot法分别检测细胞中环氧化酶．2（COX-2）的mRNA和蛋白表达,酶联免疫法（ELISA）检测细胞培养上清PGE2含量变化。结果RSV感染8h后即上调RAW264．7巨噬细胞COX-2mRNA转录和蛋白水平的表达。细胞培养上清中PGE2的含量逐渐增加。各指标的变化与正常对照相比差异均有显著性,并且随着RSV感染时间的延长而表达量增加。结论RSV感染RAW264．7巨噬细胞早期即可诱导COX-2的表达,同时生成大量的PGE2,PGE2的合成主要受COX-2的调节,COX．2参与了RSV感染所致的炎症反应。     

腹腔镜胆囊切除术的术前术后健康教育

胆结石合并胆囊炎为常见急腹症。胆囊黏膜受胆囊内结石的刺激而发生慢性炎性反应,嵌顿在胆囊颈部或胆囊管的结石,因继发感染而导致胆囊发生炎症。其发生的主要因素有胆汁淤滞、细菌感染和胆汁成分的改变,这3种因素有着内在联系,可互相影响、互为因果。胆结石可分为3类:胆色素结     

胆囊胆固醇结石患者肝脏羧肽酶E表达

目的 分析胆囊胆固醇结石患者肝脏羧肽酶E(CPE)的表达,探讨胆囊胆固醇结石形成的可能机制.方法 肝胆手术留取肝活检标本30例.其中,胆囊胆固醇结石20例(结石组),胆囊无结石患者10例(对照组).利用qRT-PCR、蛋白免疫印迹法测定两组肝脏CPE表达,用ELISA法测定血清胆囊收缩素(CCK)表达;并测定血清总胆固醇及血清总三酰甘油含量,分析胆汁成分及胆固醇饱和指数(CSI).结果 结石组胆汁总胆固醇、胆汁总脂含量及CSI均高于对照组(P＜0.05),结石组血清CCK、肝脏CPE mRNA表达及CPE蛋白表达均低于对照组(P＜0.05).结石组患者血清CCK含量与肝脏CPE mRNA表达呈正相关(r2=0.710,P＜0.05).结论 肝脏CPE表达降低可能导致血清C CK分泌减少,进而诱发结石形成.     

小白菊内酯对结肠癌细胞凋亡的影响及其机制研究

目的观察小白菊内酯（PN）对结肠癌HT-29细胞凋亡的影响并分析其可能机制。方法不同浓度PN作用于结肠癌HT-29细胞,以流式细胞仪（FCM）检测细胞凋亡;Western Blot法检测核转录因子（NF）-B亚单位pso、环氧化酶（cox-2）蛋白表达的改变;ELISA法测定前列腺素E2（PGE2）浓度。结果PN能诱导结肠癌细胞的凋亡,且随PN浓度的增加,结肠癌HT-29细胞的凋亡率增加;而NF-κB亚单位p50、COX-2蛋白表达减少,同时PGE2浓度降低。结论PN可诱导结肠癌HT-29细胞凋亡,其机制可能与其能下调NF-κB、COX-2蛋白表达并减少PGE2生成有关。     

加味茵陈蒿汤治疗胆色素结石的实验研究

目的在基础实验层面研究加味茵陈蒿汤对胆色素结石的疗效,进而为临床上肝内胆管结石的治疗探讨一种比较安全有效的方法。方法利用随机分组对照法将实验动物分为5组(正常对照组、模型组、小剂量治疗组、中剂量治疗组、大剂量治疗组)给予不同的干预措施,包括皮下注射洁霉素制造豚鼠胆色素结石模型和加味茵陈蒿汤灌胃,实验结束,测定血清中总胆红素(TBIL)、直接胆红素(DBIL)、血清钙(Ca)、丙氨酸转氨酶(ALT)的水平,并用SPSS10.0统计软件进行处理。结果各所测指标在正常组与模型组之间及模型组与各治疗组间相比差异有统计学意义(P<0.05)。结论加味茵陈蒿汤能有效去除豚鼠胆色素结石,降低血清中胆红素水平,有效保护肝功能,尤以中剂量组最为明显。     

舒胆通颗粒防治胆石症的实验研究

目的：观察舒胆通颗粒对豚鼠实验性胆石症的防治作用。方法：62只DHA系豚鼠随机分为正常组、模型对照组、阳性对照组（复方胆通胶囊,0.6 g/kg）以及舒胆通颗粒5、10、20 g（按生药量折算,下同）/kg剂量组,除正常组外均采用高胆固醇低蛋白致石饲料喂养复制胆石症模型,同时灌胃给药连续70 d,观察豚鼠一般状况、结石形成、胆汁分泌及其成分、血液生化等方面的变化。结果：舒胆通颗粒5、10、20 g/kg剂量能明显改善胆石症豚鼠一般状况,降低结石形成比率,提高胆汁中总胆汁酸（TBA）含量,10、20 g/kg并能明显提高胆汁分泌量以及血浆胆囊收缩素（CCK）含量,降低血清TC水平。结论：舒胆通颗粒对胆石症豚鼠模型有较好的防治作用。     

肝胆结石片的体外溶石、抑菌、解痉作用

试验表明：5％、5．5％的肝胆结石片药液溶石10天、对人体胆色素结石、胆固醇结石、混合型结石的溶石率分别为12．4％、6．2％；9．0％、6．0％；19．5％、10．0％。45天的溶石率分别为91．0％、57．4％；15．1％、10．1％；90．5％、60．0％。对胆色素、混合型两种结石的溶石作用显著。抑菌试验表明：对金黄色葡萄球菌、肺炎球菌、（甲、乙）型溶血性链球菌及沙门氏菌的MIC分别为0、0078、0．0156、0.0156、0．0156及0．03259／ml。1％或1‰浓度的药液对1‰BaCl、1‰。000Ach、1‰。005-HT所致豚鼠肠管痉挛均有明显的解控作用，提示本品对平滑肌有一定的松弛作用。     

Significant differences in the disposition of cyclic prodrugs of opioid peptides in rats and guinea pigs following IV administration

The stabilities of DADLE ([D-Ala2,D-Leu5]-Enk, H-Tyr-D-Ala-Gly-Phe-D-Leu-OH), the capped derivative Ac-DADLE-NH2, and the oxymethyl-coumarinic acid (OMCA)-based cyclic prodrug of DADLE and [D-Ala2,Leu5]-Enk (H-Tyr-D-Ala-Gly-Phe-Leu-OH) were determined at 37 degrees C in rat and guinea pig liver microsomes in the presence and absence of paraoxon, an esterase B inhibitor, and ketoconazole, a CYP3A4 inhibitor. These studies showed that the order of stability in microsomes was: DADLE > Ac-DADLE-NH2 > OMCA-DADLE = OMCA-[D-Ala2,Leu5]-Enk. While paraoxon produced no significant effect on the stability of the studied compounds in liver microsomes, ketoconazole inhibited the metabolism, suggesting that the capped peptide and the cyclic prodrugs are substrates for cytochrome P450 enzymes. For pharmacokinetic studies, the cyclic prodrugs of DADLE and [D-Ala2,Leu5]-Enk were administered i.v. to rats and guinea pigs. Various biological fluids and tissue (brain, bile, and blood) were collected and analyzed for the free peptide and the prodrugs by high performance liquid chromatography with tandem mass spectrometric detection (LC-MS-MS). These studies showed that the conversion of the cyclic prodrugs to the respective linear peptides (i.e., DADLE and [D-Ala2,Leu5]-Enk) was rapid in rat and guinea pig. In terms of drug elimination, only trace amounts of OMCA-DADLE and OMCA-[D-Ala2,Leu5]-Enk were recovered in guinea pig bile (3.3% and 0.82%, respectively), while significant amounts were recovered in rat bile (38.1% and 51.7%, respectively). Brain uptake of the cyclic prodrugs in guinea pigs compared to previously determined brain uptake of OMCA-DADLE in rats was also significantly different. For OMCA-DADLE, the brain levels of the cyclic prodrug and DADLE in guinea pigs were approximately 80 and 8.5 times greater, respectively, than the levels observed in rat brain. The brain-to-plasma prodrug concentration ratios in guinea pigs (>or= 0.6) were significantly higher than the ratio observed in rats (0.01). These species differences are most likely due to the different substrate specificities of the efflux transporters that facilitate liver clearance of these prodrugs and limit their permeation into the brain.     

鼻敏片对寒证变应性鼻炎豚鼠辅助性T细胞17/调节性T细胞平衡的影响

目的:探讨鼻敏片对寒证变应性鼻炎(allergic rhinitis,AR)豚鼠辅助性T细胞17(Th17)/调节性T细胞(Treg)表达水平的影响及其作用机制.方法:将40只雄性豚鼠随机分为空白组、模型组、鼻敏片组和氯雷他定片组,以复方凉药+卵清蛋白致敏法建立寒证AR模型后给予相应的药物干预,治疗15 d后,取血清和...     

雷公藤对豚鼠哮喘模型核因子-κB表达的作用

目的　探讨雷公藤对核因子 κB(NF κB)在豚鼠哮喘模型中表达的影响。方法　豚鼠用 10 %卵白蛋白 (OA)溶液1ml腹腔注射致敏 ,2wk后用 1%OA溶液超声雾化吸入致其哮喘发作 ,每日 1次 ,共 1wk ;测定肺功能并观察气道壁嗜酸性粒细胞 (EOS)浸润情况 ;用免疫组织化学染色方法观察NF κB在豚鼠肺组织中的表达变化。结果　NF κB主要表达在气道壁内的细胞 ,对照组、模型组、雷公藤内酯醇 ( 10 0mg·kg-1ip)组的胞核阳性的细胞数占气道上皮细胞数百分比分别为 14 3 %± 2 6%、3 2 5 %± 5 8%和 19 7%±2 1%。结论　雷公藤内酯醇能显著抑制哮喘豚鼠气道壁内细胞NF κB的表达 ,可能是雷公藤治疗哮喘的作用机制之一。     

乳舒片对乳腺增生模型豚鼠的实验研究

目的:观察乳舒片对乳腺增生模型豚鼠的治疗作用。方法:采用苯甲酸雌二醇联合黄体酮肌注的方法.复制豚鼠乳腺增生模型,用乳舒片治疗30 d,测定豚鼠乳头直径及高度;测定豚鼠血清雌二醇(E_2)和孕酮(P)的含量。结果:乳舒片组豚鼠乳腺高度及乳腺直径均较模型对照组明显缩小,显著降低豚鼠血清E_2和P水平。结论:乳舒片对豚鼠乳腺增生痛有较好的疗效。     

Species differences in disposition of benzo[a]pyrene

Comparison of disposition of benzo[a]pyrene (B[a]P) among Sprague-Dawley rats, Gunn rats, hamsters, and guinea pigs was performed. [3H]B[a]P was administered intratracheally to animals, and the rate of excretion of radioactivity into bile, types of metabolites of B[a]P in bile, and distribution of radioactivity among tissues were determined. In Sprague-Dawley rats, Gunn rats, and guinea pigs, the rate of excretion of radioactivity was dependent upon the administered dose. Excretion and tissue distribution of radioactivity were qualitatively similar among these species although quantitative differences were observed. In hamsters, the rate of excretion was essentially independent of dose at the concentrations examined (0.16 and 350 micrograms). The major difference between hamsters and the other species was that increased amounts of radioactivity were retained in lungs of hamsters at the lower dose with a proportional decrease in the amount of radioactivity excreted into bile. The types and relative amounts of conjugated and nonconjugated metabolites of B[a]P were similar in bile of Sprague-Dawley rats and hamsters. Smaller amounts of glucuronides and larger amounts of sulfate conjugates were detected in bile of Gunn rats than in bile of Sprague-Dawley rats or hamsters. Metabolites in bile of guinea pigs were markedly different from those in the other species in that approximately 90% of the metabolites were thioether conjugates. Buthionine sulfoxime was used to reduce tissue levels of glutathione in Sprague-Dawley rats. When liver and lung glutathione levels were reduced to 30% and 82% of control levels, respectively, the amount of radioactivity excreted into bile was not significantly different from controls.(ABSTRACT TRUNCATED AT 250 WORDS)     

Species difference in biliary excretion of methylmercury

Species difference in the biliary excretion of methylmercury was studied in male rats, mice, rabbits and guinea pigs. The rates of mercury excretion (% dose/2 hr) into the bile of the rats, mice, rabbits and guinea pigs during the 2 hr from 2 to 4 hr after the administration of methylmercury were 0.61, 0.091, 0.036 and 0.019, respectively. These results suggest that biliary excretion and enterohepatic circulation of methylmercury in the latter three species may not influence the fate of this compound as significantly as in rats. Most of the methylmercury excreted into the bile of rats was bound to glutathione (GSH). In the mouse bile, 40% of the methylmercury was bound to GSH and the rest was found in a fraction eluted at the void volume of the Sephadex G-15 column. However, in the case of the rabbits and guinea pigs, methylmercury-GSH was scarcely detectable in the bile and almost all of the methylmercury was eluted at the void volume of the column.     

热喘平口服液对豚鼠哮喘模型血清炎性介质影响的实验研究

目的:探讨热喘平口服液对豚鼠哮喘模型的平喘作用机制.方法:将50只清洁级封闭群幼龄雄性豚鼠随机分成5组,即哮喘模型组、中药小剂量组、中药大剂量组、地塞米松组、正常对照组,每组10只.除正常对照组外,其余4组采用腹腔注射10%卵白蛋白(OVA)造模,造模成功后分别按要求给药,用药7 d后取血清,检测血清中神经源性气道炎性介质P物质(SP)、血栓素B2(TXB2)、白三烯B4(LTB4)、6-酮-前列腺素F1α (6 keto PGF1α)的含量.结果:与哮喘模型组比较,中药大、小剂量组均能降低哮喘豚鼠血清中SP,TXB2,LTB4含量(P<0.05或P<0.01),升高6 keto PGF1α水平(P<0.05或P<0.01),并且中药大、小剂量组之间存在量效关系(P<0.05).中药大剂量组SP,TXB2,LTB4及6 keto PGF1α与地塞米松组比较,差异均无统计意义(P>0.05),说明大剂量热喘平口服液与地塞米松疗效相当.结论:热喘平口服液能降低血清中SP,TXB2,LTB4含量,升高6 keto PGF1α水平,提示该药的作用机制与调节血清炎性介质有关,从而发挥解痉、平喘的药理效应.