罗格列酮对低氧性肺动脉高压大鼠PPARγ和PTEN表达的影响

目的:研究罗格列酮(rosiglitazone,RSG)对低氧性肺动脉高压大鼠过氧化物酶体增殖物激活受体γ(Peroxisome proliferator activated receptor gamma,PPARγ)和10号染色体缺失张力蛋白同源磷酸酶基因(Phosphatase and tensin homolog deleted on chromosome 10,PTEN)表达的影响。方法:SD大鼠随机分为正常对照组、低氧组、低氧+罗格利酮组,建立低氧性肺动脉高压大鼠模型,4周后测定各组大鼠右心室压力、右心肥厚指标,同时检测各实验组PPARγ、PTEN的表达和组织病理学变化。培养原代大鼠肺动脉平滑肌细胞,分别给与低氧、低氧+罗格列酮、低氧+GW9662处理后观察细胞增殖及PPARγ、PTEN的表达变化。结果:1与正常组相比,低氧组大鼠右心室压力、右心肥厚指标明显增加,肺小动脉管壁增厚,PPARγ、PTEN的表达明显减少。与低氧组相比,低氧+罗格列酮组大鼠右心室压力下降,右心室及肺小动脉管壁的肥厚减轻,PTEN的表达增加。2低氧下,PASMCs中PPARγ、PTEN表达明显减低,细胞增殖较常氧明显增加,给与罗格列酮后,PTEN表达增加,给与GW9662,PTEN表达减少。3罗格列酮可以抑制PASMCs低氧下的增殖,而给与GW9662后,这一抑制作用减轻。结论:早期应用罗格列酮可激活低氧性肺动脉高压大鼠PPARγ的活性,进而上调PTEN表达,改善低氧性肺动脉高压。     

CXCR4 抑制剂AMD3100 对大鼠低氧性肺动脉高压作用的实验研究

目的:研究CXC趋化因子受体-4(CXC Chemokine receptor-4,CXCR4)的抑制剂(AMD3100)对大鼠低氧性肺动脉高压的影响。方法:将实验动物随机分为常氧对照组、低氧组、低氧+AMD3100组,采用低压低氧法建立大鼠低氧性肺动脉高压模型,4周后观察低氧对CXCR4表达的影响及各组大鼠血流动力学、右心室肥厚指标和组织病理学改变。培养原代大鼠肺动脉平滑肌细胞(Pulmonary arterial smooth cells,PASMCs),分别低氧处理及给予AMD3100,观察细胞迁移、增殖情况。结果:1低氧组大鼠CXCR4表达增加,右心室压力(Mean right ventricle pressure,m RVP)、右心室肥厚指标(Right ventricle/Body weight,RV/BW;Right ventricle/Left ventricle plus septum,RV/(LV+S))增加,肺细小动脉管壁增厚,造模成功;低氧+AMD3100组大鼠m RVP和RV/BW、RV/(LV+S)比值、肺细小动脉管壁增厚程度较低氧组明显降低(P0.05)。2低氧组PASMCs与常氧组相比,细胞迁移及增殖均明显增加;AMD3100组PASMCs迁移和增殖与低氧组相比受抑制(P0.05)。结论:AMD3100能有效的降低大鼠低氧性肺动脉高压的m RVP,抑制肺细小动脉管壁的增生,减轻右心室的肥厚,其有可能是通过抑制了PASMCs的迁移和增殖,从而抑制肺血管的重建,防治低氧性肺动脉高压。     

肝素通过增加细胞周期抑制因子p27 减轻大鼠低氧性肺动脉高压

目的:研究肝素防治大鼠低氧性肺动脉高压的作用机制。方法:复制慢性低氧性肺动脉高压大鼠模型,将大鼠分为常氧对照组(Hypoxia),常氧加肝素组(Normoxia+H),缺氧模型组(Hypoxia)及缺氧加肝素组(Hypoxia+H)共四组。待模型复制完毕后,测定大鼠RVPSP和RV/LV+S,然后进行组织病理分析,测定MA%和MT%以确定肺动脉高压的程度。利用Western-blot实验与RT-PCR实验检测肺动脉中p27蛋白水平及RNA水平的变化。结果:慢性低氧显著增加了大鼠RVPSP、RV/LV+S、MA%和MT%,引起了大鼠肺动脉压力的增高、右心室肥厚和肺中小动脉的中膜增厚及外膜增生,肝素的使用减轻了慢性缺氧条件下大鼠的RVPSP和RV/LV+S,降低了MA%和MT%,有效降低了肺动脉高压、右心室肥厚,并减轻了肺中小动脉的中膜增厚和外膜增生。Western-blot实验发现使用肝素的缺氧组大鼠肺动脉中p27蛋白表达增加,而RT-PCR实验发现p27的转录水平变化不显著。结论:结果显示肝素可能通过增加p27的表达,从而抑制肺动脉平滑肌细胞的增殖,减轻了肺动脉血管的狭窄及降低了肺动脉高压。     

培美曲塞联合罗格列酮抑制肺癌A549 细胞增殖研究

目的:研究培美曲塞联合过氧化物酶体增殖物激活受体γ(Peroxisome proliferators-activated receptorγ,PPARγ)激动剂罗格列酮(Rosiglitazone,RSG)对人肺癌A549细胞株增殖能力的影响。方法:蛋白质印迹法检测肺癌细胞系中PPARγ的表达水平,筛选高表达PPARγ的肺癌细胞株利用该细胞株并分别给予培美曲塞、培美曲塞联合罗格列酮、培美曲塞联合罗格列酮及PPARγ拮抗剂GW9662处理,之后检测细胞增殖能力及PPARγ、PTEN、pAKT表达水平的变化。结果:PPARγ在A549细胞系中显著高表达;培美曲塞组、培美曲塞联合罗格列酮组、培美曲塞联合罗格列酮及GW9662组,肺癌细胞增殖均受到抑制,吾美曲塞和罗格列酮联合应用对肺癌细胞的增殖抑制具有协同效应,与单用培美曲塞组相比有统计学差异,且该效应可被GW9662有效阻断;给予培美曲塞和罗格列酮联合应用,可明显上调PPARγ、PTEN表达,下调pAKT表达;给予PPARγ拮抗剂GW9662后,PPARγ、PTEN的表达显著下调,pAKT表达上调。结论:PPARγ激动剂RSG对培美曲塞抑制肺癌细胞的增殖具有协同效应且其分子机制可能是通过激活PPARγ/PTEN/pAKT信号通路从而抑制肺癌A549细胞增殖。     

肺动脉高压大鼠肺内5-HT1B受体的分布和表达变化

目的:观察低氧性肺动脉高压大鼠肺内5-HT1B受体的分布和表达变化,探讨低氧性肺动脉高压的形成机制.方法:40只健康雄性SD大鼠随机分为正常组(control)、低氧3周组(2w)、低氧4周组(4w)和低氧5周组(5w).除正常组外,其余3组大鼠分别在低氧环境中饲养3周、4周和5周.测定各组大鼠的平均肺动脉压力(mPAP)、右心室收缩压(RVSP)和右心室肥厚度[RV/(LV+S)%].应用免疫组织化学法观察大鼠肺组织中5-HT1B受体的分布和表达,Western blot法测定大鼠肺组织中5-HT1B受体的蛋白含量.结果:和正常组相比,低氧3周组大鼠的mPAP、RVSP和右心室肥厚度均显著升高(P均＜0.05),并且随着低氧时间的延长而持续升高(P均＜0.05).免疫组织化学结果显示:5-HT1B受体主要分布在正常大鼠肺动脉的内膜层,而平滑肌层中仅有少量表达:和正常组相比,低氧3周组大鼠肺动脉平滑肌层中5-HT1B受体的表达显著增多;随着低氧时间的延长,大鼠肺动脉平滑肌层中5-HT1B受体表达持续增多.Western blot结果表明,大鼠肺组织中5-HT1B受体的蛋白含量变化和免疫组织化学结果相一致.结论:低氧性肺动脉高压大鼠肺动脉中5-HT1B受体呈过度表达,这可能是低氧性肺动脉高压形成的分子机制之一.     

低氧对大鼠右心室肥厚及心肌中缝隙连接蛋白43表达的影响

目的:研究低氧对大鼠右心室肥厚及心肌中缝隙连接蛋白43 (Cx43)表达的影响.方法:40只健康雄性SD大鼠随机分为正常组(control)、低氧3周组、低氧4周组和低氧5周组.除正常组外,其余3组大鼠分别在低氧环境中饲养3周、4周和5周.测定和比较各组大鼠的平均肺动脉压力(mPAP)、右心室收缩压(RVSP)、右心室肥厚度[Rv/(LV+S)％],并通过免疫组化染色法观察各组大鼠左心室心肌细胞中cx43的表达.结果:与正常组相比,低氧3周、4周、5周组大鼠的mPAP、RVSP、右心室肥厚度均显著升高(P均＜0.05),与低氧3周组比较,低氧4周、5周组大鼠的mPAP、RVSP、右心室肥厚度均显著升高(P均＜0.05),而低氧5周组大鼠的mPAP、RVSP、右心室肥厚度均显著高于低氧4周(P均＜0.05).免疫组织化学结果显示:低氧组大鼠Cx43排列紊乱,端-端连接减少,侧面连接增多;随着低氧时间的延长,大鼠心肌细胞中Cx43的表达逐渐减少,差异具有统计学意义(P均＜0.05).结论:低氧可导致右心室肥厚,并随着诱导时间的延长而逐渐加重,这可能与心肌中Cx43的分布紊乱及表达减少有关.     

慢性低氧性肺动脉高压大鼠肺组织内质网应激介导的凋亡的变化

目的:探讨内质网应激介导的凋亡在低氧性肺动脉高压大鼠肺组织中的变化及意义.方法:清洁级雄性SD大鼠22只随机被均分成对照组和低氧组(n=11).采用常压低氧法复制慢性低氧高二氧化碳性肺动脉高压模型,4周后,测定肺动脉平均压(mPAP)、右心室游离壁(RV)和左心室加室间隔(LV+S)重量比、肺细小动脉管壁面积/管总面积...     

RhoA/ROCK信号通路在左心疾病致大鼠肺动脉高压模型中的作用

目的探讨Rho A/ROCK信号通路在左心疾病相关的大鼠肺动脉高压模型中的表达水平和作用。方法 3~4周龄雄性SD大鼠20只,体重90~100 g,随机分为对照组(C组:n=10)、肺高压组(H组:n=10)。H组采用升主动脉固定缩窄术建造左心疾病相关肺动脉高压大鼠模型,C组大鼠行假手术处理(钛夹固定于血管旁纵隔组织而非升主动脉,其他所有手术操作同H组),在建模后60 d,对各组大鼠进行血流动力学(右心室收缩压、肺动脉压力)检测,处死大鼠并用PBS行在体心肺灌洗致双肺变白,左肺组织固定于4%多聚甲醛行病理切片以观察肺组织病理形态学变化、右肺组织冻存以备生物分子学检测(Rho激酶mRNA、Rho A mRNA、ET-A受体mRNA)。结果与C组相比,H组肺动脉压力、右心室收缩压明显增高(P0.01),肺小动脉壁明显增厚,肺小动脉管腔狭窄甚至闭塞,管壁肥厚指数明显增大(P0.01);与C组相比,H组肺组织的Rho激酶mRNA表达水平明显增加,Rho A mRNA、ET-A受体mRNA表达水平亦明显增加,差异均有统计学意义(P0.01)。结论采用升主动脉固定缩窄术成功建造了左心疾病相关肺动脉高压大鼠模型;与C组相比,H组肺小血管壁明显增厚,肺组织的Rho激酶mRNA、Rho A mRNA、ET-A受体mRNA表达明显增高,该信号通路可能参与了左心疾病相关肺动脉高压的形成过程。     

尾加压素Ⅱ及其受体在肺动脉高压大鼠右心室的表达

目的：观察尾加压素Ⅱ（UⅡ）及其受体UT在慢性低Q高C02肺动脉高压大鼠右心室的表达。方法：健康SD大鼠20只随机分成正常对照组和低氧高二氧化碳4周（HH）组,分别测定平均肺动脉压力（InPAP）,右心室游离壁（Rv）和左心室加室间隔（LV＋S）的重量比;放免法测定大鼠血浆UⅡ含量;免疫组化方法检测心肌细胞、心肌小动脉UⅡ蛋白的表达;组织原位杂交方法检测心肌细胞、心肌小动脉UⅡmRNA和UⅡ受体（UT）mRNA的表达。结果：①HH组InPAP和RV／LV＋S比正常对照组分别高52．0％与25．4％（P均〈0．01）。②HH组血浆UⅡ水平较正常对照组无明显增高。③免疫组化显示HH组心肌细胞UⅡ蛋白表达阳性率和心肌小动脉UⅡ蛋白表达的平均吸光度值均明显高于正常对照组（P〈0．01）。④组织原位杂交可见HH组的心肌细胞UⅡmRNA表达阳性率和心肌小动脉UⅡmRNA表达的平均吸光度值均明显高于正常对照组（P均〈0．01）。⑤组织原位杂交可见HH组的心肌细胞UTmRNA表达阳性率和心肌小动脉UTmRNA表达的平均吸光度值均较正常对照组明显增高（P均〈0．01）。结论：慢性低氧高二氧化碳性大鼠肺动脉高压及右室肥大形成过程中,心肌小动脉和心肌细胞的UⅡ及其受体UT的表达均呈现明显上调,提示UⅡ可能有促进心肌细胞增殖,导致右心室重构的作用。推测UⅡ在慢性低氧高二氧化碳性肺动脉高压及右室肥大的形成机制中具有重要的病理生理意义。     

一种新型PPARr激动剂对人肾癌细胞增殖抑制的实验研究

目的：探讨新型过氧化物酶体增殖激活物受体(PPARr)激动剂DH9 对人肾癌细胞OS-RC-2 的增殖抑制作用。方法：予以不 同浓度的DH9 及罗格列酮作用OS-RC-2 细胞12 h、24 h和48 h,荧光素酶活性检测比较两种药物的PPARr激动效应;MTT 法 检测细胞增殖情况;流式细胞术观察细胞周期;AnnexinV-FITC/PI双染色流式细胞术测定细胞凋亡率;Western blot 检测细胞内 Bax 及Bcl-2等蛋白的变化。结果：不同浓度的DH9 与罗格列酮相比,对PPARr的激动效应DH9明显低于罗格列酮,增殖抑制 作用优于罗格列酮(P＜0.05),并呈现明显的浓度、时间依赖性;加入PPARr抑制剂GW9662 前后DH9 的增殖抑制作用差异无统 计学意义(P＞0.05);DH9 作用细胞48小时后,G0/G1 期细胞比例明显增加(P＜0.05),S期细胞明显减少(P＜0.05)。DH9可诱导细 胞凋亡,伴随Bcl-2 表达的减少以及Bax表达的增加。结论：OS-RC-2 细胞中,DH9 的增殖作用明显优于罗格列酮,且是通过 PPARr非依赖途径实现;DH9 能将OS-RC-2 细胞阻滞在G0/G1 期,并通过影响Bcl-2 和Bax 蛋白表达促进细胞凋亡。     

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.     

真菌类遗传学分析的知识结构教学

本文以认知结构理论为指导,讨论了真菌类遗传分析与高等动植物遗传分析的内在联系,认为利用这种内在联系进行教学可收到好的效果并说明了作者的具体教学过程。 Abstract:In the paper, the relationship between genetic analysis of Fungi and genetic analysis of high animal and plant was discussed.A good results were obtained when we adopted this method in the teaching.     

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.