Sunitinib对哮喘气道重塑小鼠PDGFR-β磷酸化和MMP-9表达的影响

目的:探讨sunitinib对支气管哮喘气道重塑的干预作用及可能的作用机制.方法:BALB/c小鼠随机分为正常对照组、哮喘气道重塑组、sunitinib干预组.鸡卵清蛋白致敏和激发建立哮喘小鼠气道重塑模型;收集支气管肺泡灌洗液(BALF)做细胞计数;取右肺组织行苏木精-伊红(HE)染色和Masson三色染色;免疫沉淀(IP法)测定小鼠肺组织PDGFR-β受体磷酸化及westernblot(WB)法检测MMP-9蛋白质的表达.结果:HE和Masson三色染色提示哮喘组黏膜下层和平滑肌增厚、气道管腔狭窄、胶原纤维增生、大量炎症细胞浸润,sunitinib干预组上述改变较哮喘组为轻;哮喘组BALF中炎症细胞总数和嗜酸性粒细胞(EOS)计数在哮喘组表达较对照组为高(P＜0.05),而sunitinib组低于哮喘组(P＜0.05);PDGFR-β受体的磷酸化和MMP-9的表达在哮喘组表达较对照组为高(P＜0.01),而sunitinib干预组表达量低于哮喘组(P＜0.01).结论:sunitinib能够抑制哮喘小鼠PDGFR-β的磷酸化和MMP-9表达,减轻气道炎症反应、延缓气道重塑进程.     

核心蛋白聚糖对慢性哮喘小鼠气道和肺血管胶原沉积的影响

目的:研究核心蛋白聚糖(decorin)对哮喘小鼠气道及肺血管胶原沉积的影响.方法:30只雌性昆明系小鼠随机分为对照组、慢性哮喘组及decorin干预组,每组10只.慢性哮喘组和decorin干预组小鼠第0、7、14d腹腔注射卵蛋白(OVA)致敏;第22d始雾化吸入2.5％OVA溶液激发哮喘,每次30min,每周3次,连续8周;decorin干预组小鼠每次OVA激发前2h腹腔注射decorin(0.25mg·kg-1)干预;对照组全部以生理盐水代替.末次激发24h后处死小鼠.酶联免疫吸附测定(ELISA)法测定小鼠血清和肺泡灌洗液(BALF)中转化生长因子-β1 (TGF-β1)水平;苏木精-伊红(HE)染色及Masson三色染色观察肺组织切片病理改变及胶原沉积;图像分析软件测定气道血管胶原沉积.结果:慢性哮喘组血清及BALF中TGF-β1水平显著高于对照组(P均＜0.01),而decorin 干预组较慢性哮喘组明显降低(P均＜0.01);慢性哮喘组气道壁胶原沉积厚度及肺血管壁胶原沉积厚度显著高于对照组(P均＜0.01),而decorin干预组上述改变较慢性哮喘组明显减轻(P均＜0.01).结论:Decorin干预可减轻哮喘小鼠气道及肺血管胶原沉积.     

婴儿型双歧杆菌对哮喘小鼠气道炎症的作用研究

目的探讨婴儿型双歧杆菌对气道过敏小鼠的气道炎症作用及其机制。方法 6周龄BALB/c雄性小鼠40只,随机分为4组:阴性对照组、哮喘组、婴儿型双歧杆菌预防组和婴儿型双歧杆菌治疗组。哮喘组、预防组和治疗组用卵清蛋白(Ovalbvmin,OVA)进行致敏和激发诱发哮喘,阴性对照组用生理盐水进行致敏和激发。预防组第0至14天灌胃菌液;治疗组第15至28天灌胃菌液;阴性对照组和哮喘组全程灌胃生理盐水作对照。观察小鼠气道过敏表现;计数肺泡灌洗液(Bronchoalveolar lavage fluid,BALF)中细胞总数评估气道炎症程度;肺组织行HE染色;ELISA法测定血清中IL-10、OVA特异性IgE和OVA特异性IgG1的浓度以及肺泡灌洗液中细胞因子IL-4、IL-5、IL-10、IL-13和IFN-γ的浓度。结果 (1)哮喘组小鼠在激发后出现搔抓口鼻、竖毛、打喷嚏、腹肌扇动、弓背直立、二便失禁以及体重下降等表现,预防组和治疗组的症状较轻。(2)预防组和治疗组BALF细胞数量明显低于哮喘组(P0.05)。(3)肺组织HE染色后,哮喘组小鼠肺组织出现明显的炎症细胞浸润,预防组和治疗组肺组织炎症明显减轻。(4)预防组和治疗组血清OVA特异性IgE浓度明显低于哮喘组(P0.05),并且预防组低于治疗组(P0.05);治疗组血清OVA特异性IgG1浓度明显低于哮喘组和预防组(P0.05);预防组和治疗组血清IL-10浓度明显高于哮喘组(P0.05),且治疗组高于预防组(P0.05);(5)预防组和治疗组肺泡灌洗液中IL-4、IL-13含量明显低于哮喘组(P0.05);IL-5、IL-10、IFN-γ各组浓度都较低,未测出有效浓度。结论口服婴儿型双歧杆菌可以减轻过敏症状,降低肺组织炎症浸润程度,抑制Th2免疫反应。     

通光散汤对小鼠哮喘模型气道高反应性和气道炎症的影响

目的观察通光散对小鼠哮喘模型气道反应和气道炎症的影响。方法35只6周龄BALB／c小鼠随机分为哮喘模型组、正常对照组和药物实验组。模型组和药物组以鸡卵白蛋白（OVA）致敏、激发;药物组在最后一次致敏后每天灌胃给予通光散汤0．72mL（相当于0．04g生药）;对照组以等体积的Ns代替OVA致敏、激发。末次激发48h后处理小鼠：无创法测定小鼠的气道高反应性,观察气道阻力变化;支气管肺泡灌洗液（BALF）行细胞学分类;观察肺组织的病理变化。结果①药物组小鼠气道阻力的变化与模型组相比明显下降,差异显著（P＜0．05）;②药物组BALF白细胞总数和Eos（％）与模型组相比明显降低（P＜0．05）。③模型组小鼠肺脏组织支气管、血管黏膜下和周围肺组织有明显的炎症细胞浸润,大量炎症细胞向支气管和血管迁移,上皮细胞部分有脱落,部分可见黏液栓,血管壁明显水肿;治疗组小鼠肺组织炎性细胞浸润和管腔黏液分泌情况较模型组明显减轻,气道粘液的分泌量得到明显的控制。结论通光散汤对小鼠哮喘模型气道高反应性和气道炎症有显著抑制作用。     

口服柔嫩梭菌对OVA致敏小鼠气道炎症的影响

目的 在动物水平探索口服柔嫩梭菌(Clostridium leptum)对哮喘小鼠气道炎症的影响.方法 建立OVA致敏的BALB/c小鼠哮喘模型,依据检测气道炎症情况和气道反应性确定哮喘模型构建成功.30只BALB/c小鼠随机分为3组:正常对照组,安慰剂组和口服柔嫩梭菌治疗组,每组10只.通过HE染色检测小鼠肺组织病理变化,细胞计数检测肺泡灌洗液中炎性细胞(嗜酸性粒细胞、中性粒细胞、淋巴细胞、巨噬细胞)的数目,ELISA方法检测肺泡灌洗液中炎性因子(IL4、IL-5、IL-13)的表达情况,并检测各组小鼠的气道反应性.结果 验证OVA致敏哮喘小鼠模型构建成功;口服柔嫩梭菌可显著减轻OVA致敏小鼠的气道高反应性,气道炎症细胞浸润和炎性细胞因子分泌(P＜0.05).结论 口服柔嫩梭菌可显著减轻OVA致敏小鼠的气道炎症和气道高反应性,这可能为哮喘治疗提供新思路.     

木犀草素对哮喘小鼠转录因子GATA-3表达的影响

目的:观察木犀草素对哮喘小鼠转录因子GATA-3表达的影响.方法:将30只BALB/c小鼠随机分为正常对照组、哮喘气道重塑组、木犀草素干预组,每组10只;鸡卵清蛋白致敏和激发建立哮喘小鼠气道重塑模型;HE染色观察各组气道炎症发生及气道结构改变:采用Western blot及Real time PCR技术检测治疗前后哮喘小鼠肺组织中GATA-3蛋白和GATA.3 mRNA的表达.结果:HE染色提示哮喘组出现黏膜下层和平滑肌增厚,气道管腔狭窄,大量炎细胞浸润的表现,木犀草素组上述改变较哮喘组为轻:Westernblot结果显示哮喘组小鼠肺组织GATA-3蛋白表达较对照组为高(P＜0.01),木犀草素治疗组表达量低于哮喘组(P＜0.01).Real timePCR结果与Westernblot结果一致.结论:木犀草素干预可下调哮喘引起的GATA一3的表达;木犀草素抑制GATA-3表达可能是其缓解气道炎症的机制之一.     

咪喹莫特对慢性哮喘模型小鼠肺组织中基质金属蛋白酶-9-表达的影响

目的：观察Toll样受体7配体咪喹莫特对慢性哮喘小鼠模型气道重塑及肺组织中基质金属蛋白酶MMP-9表达的影响。方法：36只BALB／c小鼠按随机原则分成正常对照组、哮喘模型组、咪喹莫特组,每组12只。通过卵蛋白致敏,气道激发8周,末次激发24h后,检测各组小鼠气道反应性,HE染色观察气道炎症变化;Masson三色染色观察气道纤维化的改变;real-timePCR和western—blot分别检测肺组织中MMP-9的mRNA和蛋白表达。结果：慢性哮喘组小鼠气道炎症、气道高反应性和气道重塑较正常对照小鼠明显加重,而咪喹莫特组小鼠模型的气道炎症和气道反应性及气道重塑均较哮喘模型组小鼠减少或降低。慢性哮喘组小鼠肺组织MMP-9的mRNA和蛋白水平均较正常对照小鼠明显增加（P〈0．05）,而咪喹莫特治疗可显著降低哮喘小鼠肺组织MMP-9的mRNA和蛋白水平（P〈0．05）。结论：咪喹莫特能够显著抑制慢性哮喘小鼠模型的气道炎症、降低气道高反应性并减轻气道重塑,这可能与其抑制MMP-9的表达有关。     

小青龙汤对小鼠支气管哮喘模型气道炎症及细胞因子的影响

目的:探讨中药方剂小青龙汤对小鼠哮喘模型气道炎症及细胞因子的影响。方法:40只BALB/c小鼠随机分为正常对照组(A组)、哮喘模型组(B组)、小青龙汤低剂量组(C组)和小青龙汤高剂量组(D组)。B、C、D组采用卵蛋白(OVA)腹腔注射致敏与雾化吸入激发制作哮喘模型。于OVA激发结束后24h收集支气管肺泡灌洗液(BALF)计数炎性细胞总数及嗜酸性粒细胞(EOS)数目,并测定BALF上清液中白细胞介素-4(IL-4)和干扰素-γ(IFN-γ)水平变化。结果:小青龙汤的干预治疗能显著降低小鼠BALF中炎性细胞总数及EOS数量;BALF上清液中IFN-γ水平明显升高,IL-4水平显著下降。D、C组与A组、B组有显著性差异(P<0.05),C组与D组结果亦存在显著性差异(P<0.05)。结论:小青龙汤能明显降低哮喘小鼠BALF中炎性细胞数量,影响细胞因子水平变化,从而改善哮喘气道炎症。     

STAT3小分子抑制剂FLLL31对卵白蛋白致敏小鼠气道炎症治疗活性的初步研究

目的:研究新结构化合物FLLL31抑制卵白蛋白(OVA)致敏引发的小鼠气道炎症的活性,探讨FLLL31治疗哮喘的初步疗效。方法:雄性BALB/c小鼠(18~20 g)随机分为4组,每组10只,包括正常对照组(Control组)、模型对照组(OVA组)、地塞米松组(1 mg/kg,腹腔注射)和FLLL31组(15 mg/kg,口服灌胃)。各组小鼠分别于第0 d和第14 d致敏,每只鼠腹腔注射20μg OVA和2.25 mg Al(OH)3凝胶,FLLL31自致敏第24 d开始给药,给药周期为7 d;于第28、29、30 d以气管滴入OVA进行攻击,攻击前1 h给予受试化合物FLLL31及阳性药地塞米松。结果:小鼠肺灌流液炎症细胞分类计数、肺组织病理分析等实验结果证明,FLLL31以15 mg/kg连续给药7 d后,与模型组相比,抑制了小鼠肺部气道中炎症细胞的浸润,明显改善OVA致敏引发的小鼠气道炎症症状;ELISA结果证明FLLL31降低小鼠肺灌流液中炎症因子白细胞介素6的含量;免疫组化结果显示FLLL31降低OVA致敏小鼠肺部白细胞介素6受体浓度,减少中性粒细胞标志物淋巴细胞抗原6G(Ly6G)的表达(P0.05)。结论:特异抑制STAT3的新结构化合物FLLL31在体内对OVA致敏小鼠模型气道炎症有较好的改善和免疫调节活性。     

精氨酸甲基转移酶1和核因子кB在哮喘豚鼠中的表达变化

目的:探讨共激活因子相关的精氨酸甲基转移酶1和核因子-кB在豚鼠支气管哮喘模型气道和肺组织的表达变化.方法:24只白色雄性豚鼠随机分为:①正常对照组;②哮喘组.卵清蛋白致敏并激发后采用间接免疫荧光法检测气道及肺组织精氨酸甲基转移酶1和核因子NF-кB(P65)的表达水平,探讨其在哮喘中可能的作用机制.结果:CARM1和NF-кB(P65)在对照组和哮喘组均有阳性表达,主要在支气管-终末细支气管上皮细胞和肺组织成纤维细胞胞核表达.正常对照组和哮喘组CARM1和NF-кB(P65)的表达差异均有统计学意义.结论:在哮喘豚鼠气道上皮及肺组织CARM1和NF-кB(P65)在细胞胞核高表达,提示CARM1可能通过增强募集NF-кB到相关位点激活NF-кB信号转导通路,并启动了多种前炎性基因和免疫调节基因的转录激活从而诱发哮喘炎症反应.     

六种藓类植物茎的比较解剖学研究

通过对6种藓类植物,即褶叶青藓(Brachythecium salebrosum(Web.et Mohr.)B.S.G.)、湿地匐灯藓(Plagiomnium acutum(Lindb.)Kop.)、侧枝匐灯藓(Plagiomnium maximoviczii(Lindb.)Kop.)、大凤尾藓(Fissidensnobilis Griff.)、大羽藓(Thuidium cymbifolium(Doz.et Molk.)B.S.G.)和大灰藓(Hypnum plumaeforme Wils.)嫩茎和老茎的石蜡切片和显微观察发现,同一藓类植株的嫩茎和老茎,茎结构稳定,不同种藓类植物茎横切面具有不同特征.植物体茎横切面形状、表层细胞的层数、细胞大小和细胞壁厚薄、皮层细胞大小和形状、中轴的有无以及比例等特征可以作为藓类植物的分科分类依据之一.     

Photoregulation of seed germination of wild-type and of an aurea-mutant of tomato

Seed germination of an aurea mutant of tomato ( Lycopersicon esculentum Mill.) is promoted by continuous irradiation with red, far-red or long-wavelength far-red (758 nm) light as well as by cyclic irradiations (5 min red or 5 min far-red/25 min darkness). Far-red light applied immediately after each red does not change the germination behaviour. Seed germination of the isogenic wild-type, cv. UC-105, is promoted by continuous and cyclic red light while it is inhibited by continuous and cyclic far-red light and by continious 758 nm irradiation. Far-red irradiation reverses almost completely the promoting effect of red light. The promoting effect (in the aurea mutant) and the inhibitory effect (in the wild-type) of continuous far-red light do not show photon fluence rate dependency above 20 nmol m⁻² s⁻¹. It is concluded that phytochrome controls tomato seed germination throgh low energy responses in both the wild type and the au mutant. The promoting effect of continuous and cyclic far-red light in the au mutant can be attributed to a greater sensitivity to P_fr.     

Cause of Senescence of Nine Sorts of Flowers

The levels of endogenous phytohormones and respiratory rate in nine sorts of flowers such as Cymbidium faberi Rolfe, Nopalxochia ackermannii Kunth and others were investigated both at full bloom and senescence and meanwhile the effect of exogenous phytohormones on prolonging the blossoms and promoting ethylene production were tested. There is a high content of endogenous ethylene in all the long-lived flowere, about 3–16 folds higer than the short-lived ones. There is a high level of ABA at full blooming flowers of short-lived flowers, in which there is no or only some cytokinins in it, but the ratio of CTK (6BA+zeatin)/ABA is smaller(l.7). The endogenous ABA reached a much higher level at senescence in all nine sorts of flowers, so it is reasonable to consider that it is ABA which plays an important role of regulation in controlling flower's senescence. There is a much higher level of GA3 and zeatin in the long-lived flowers which is not demonstrated in the shortlived ones. The respiratory rate is one of the factors controtling the longevity of flowers, but it does not play a decided role. Application of 6BA and zeatin prolongs distinctly orchid’s longevity, however exogenous IAA through the promotive action on ethylene production, evidently extends the longevity of the flowers of the Nopalxochia ackermannii Kunth.     

龙胆科药用植物化学成分的研究现状

龙胆科植物在我国的分布范围很广,且多数为药用植物,其多数种属的药用植物,至今其化学成分尚未被系统研究。综述了目前龙胆科药用植物的化学成分的研究现状及一般提取方法,对近年来发现的环烯醚萜及裂环烯醚萜类化合物进行了总结,为本科药用植物的更深入研究提供了参考。     

Interconnection of parameters of regulation system of lipid peroxidation and of morphophysiological parameters of mouse liver

A complex analysis of seasonal fluctuations of the mean group parameters of the system of regulation of lipid peroxidation has been performed in liver of Balb/c mice. Association of lipid characteristics and morphophysiological parameters is studied in the Balb/c mouse liver. An inter-connection is revealed between the liver index and the amount of lysoforms of phospholipids, the scale and character of the interconnection differing essentially depending on proportion of phos-phatidylcholine in mouse liver phospholipids.     

The effects of glutamine of the maintenance of embryogenic cultures of Cryptomeria japonica

Summary Embryogenic tissues of sugi (Cryptomeria japonica) were induced on a modified Campbell and Durzan (CD) medium containing 1 μM 2,4-dichlorophenoxyacetic acid (2,4-D) and 600 mg l⁻¹ glutamine, and subcultured in the medium of the same composition for over 1 yr. This resulted in a mixed culture of embryogenic and non-embryogenic cells. When embryogenic cells were isolated and cultured independently, their capacity to form embryogenic aggregates was lost. Thus, the non-embryogenic cells present within a mixed culture system were essential to the formation of embryogenic aggregates. When embryogenic tissues were isolated and cultured independently on a high glutamine-containing (2400 mg l⁻¹) medium, dry weights and endogenous levels of glutamine increased, and the tissue could generate a large number of embryogenic aggregates. Amino acid analysis of embryogenic and non-embryogenic cells from the maintenance culture indicated a higher level of glutamine was present in the latter. The high endogenous level of glutamine in the non-embryogenic portion of mixed cell masses may be the supplier of glutamine for maintaining the embryogenic property of the tissues.     

Specificity of action of piperidylcholine derivatives as substrates of cholinesterases of various origin

The review deals with study of enzymologic properties of a novel highly specific acetylcholinesterase substrate, N-(β-acetoxyethyl) piperidinium iodomethylate (“piperidylcholine”), and its 30 derivatives that were tested as effectors of cholinesterases of mammals and various species of Pacific squids. It was proven for the first time that responsible for specificity of action was structure of cyclic ammonium grouping of the alcohol part of molecule of the ester substrate. Analysis of specificity is performed based on enzymatic hydrolysis parameters—activity of catalytic center of cholinesterases and bimolecular constant of the reaction rate that are determined at optimal and low substrate concentrations. Among the specially synthesized group of thioester compounds there is revealed one more highly specific acetylcholinesterase substrate—N-(β-acetoxyethyl) piperidinium.     

Physiological characteristics of various species of strains of carboxydobacteria

Seven strains of aerobic carbon monoxide-oxidizing bacteria (carboxydebacteria) when growing on CO as sole source of carbon and energy had doubling times which ranged from 12–42 h. The activity profiles obtained after discontinuous sucrose density gradient centrifugation indicated that the CO-oxidizing enzymes are soluble and the hydrogenases are membrane-bound in all strains examined. The CO-oxidizing enzymes of Pseudomonas carboxydohydrogena, Pseudomonas carboxydoflava, Comamonas compransoris, and the so far unidentified strains OM2, OM3, and OM4 had a molecular weight of 230,000; that of Achromobacter carboxydus amounted to 170,000. The molecular weights of the CO-oxidizing and H₂-oxidizing enzymes turned out to be identical. The cell sonicates were shown to catalyze the oxidation of both CO and H₂ with methylene blue, thionine, phenazine methosulfate, toluylene blue, dichlorophenolindophenol, cytochrome c or ferricyanide as electron acceptors. Methyl viologen, benzyl viologen, FAD⁺, FMN⁺, and NAD(P)⁺ were not reduced. The spectrum of electron acceptors was identical for all strains tested. Neither free formate, hydrogen nor oxygen gas were involved in the CO-oxidation reaction. Methylene blue was reduced by CO at a 1:1 molar ratio. The results indicate that CO-oxidation by carboxydobacteria is catalyzed by identical or similar enzymes and that the reaction obeys the equation CO+H₂OCO₂+2H⁺+2e^- as previously shown for Pseudomonas carboxydovorans.Dedicated to Otto Kandler remembering almost three decades of enjoyable cooperation     

Modulation of allostery of pyruvate kinase by shifting of an ensemble of microstates

Since the introduction of the concepts of allostery about four decades ago, much advancement has been made in elucidating the structure-function correlation in allostery. However, there are still a number of issues that remain unresolved. In this review we used mammalian pyruvate kinase (PK) as a model system to understand the role of protein dynamics in modulating cooperativity. PK has a triosephosphate isomerase (TIM)(α/β)₈ barrel structural motif. PK is an ideal system to address basic questions regarding regulatory mechanisms about this common (α/β)₈ structural motif. The simplest model accounting for all of the solution thermodynamic and kinetic data on ligand-enzyme interactions involves two conformational states, inactive E^T and active E^R. These conformational states are represented by domain movements. Further studies provide the first evidence for a differential effect of ligand binding on the dynamics of the structural elements, not major secondary structural changes. These data are consistent with our model that allosteric regulation of PK is the consequence of perturbation of the distribution of an ensemble of states in which the inactive E^T and active E^R represent the two extreme end states. Sequence differences and ligands can modulate the distribution of states leading to alterations of functions. The future work includes: defining the network of functionally connected residues; elucidating the chemical principles governing the sequence differences which affect functions; and probing the nature of mutations on the stability of the secondary structural elements, which in turn modulate allostery.