温阳消饮法对胸腔积液大鼠胸膜差异基因表达影响的实验研究

发布时间：2018-04-15 22:41

  本文选题：基因芯片技术 + 胸腔积液　； 参考：《成都中医药大学》2014年博士论文

【摘要】：目的：利用基因芯片技术,分析温阳消饮法对胸腔积液大鼠胸膜基因表达的调控作用。 方法：48只雄性SD大鼠适应性饲养3天后,随机分为A组(40只)与空白对照组(8只)。A组动物以戊巴比妥钠麻醉后,采用1%λ-角叉菜胶溶液进行右侧胸膜腔注射。24h后,运用CR数字成像系统观察胸部X光片,筛选出胸腔积液明显的大鼠,作为造模成功的动物备用。将造模成功的动物随机分为：温阳消饮组(10只),每日1次温阳消饮法代表方水煎液灌胃(5ml/kg);模型组(9只),每日1次生理盐水灌胃(5ml/kg)。空白对照组(8只),每日1次生理盐水灌胃(5ml/kg)。连续给药4天后,动物进行安乐死。每组取3只动物的右侧脏层和壁层胸膜,利用Agilent单通道表达谱芯片对胸膜组织基因表达情况进行分析。 结果： (1)壁层胸膜组织 模型组与空白对照组、温阳消饮组与模型组差异表达基因涉及多个生物学过程和信号通路。在造模因素作用下,基因Fgd4、Mef2a、Fhl3、Dyrk2、Slc24a6、 Pbrm1、LOC498368、Tp53i11、Rcan3、Hapln4、Abil和Hlx表达上调,在中药干预下表达下调；Nme7、Pold2、Klk1l和Fkbp5在造模条件下表达下调,用药后上调。 (2)脏层胸膜组织 模型组与空白对照组、温阳消饮组与模型组差异表达基因涉及多个生物学过程和信号通路。在造模因素作用下,基因Rhoa、RGD1306349、Nsd1、Figf、Brd4、 Crebbp、Sos2、Irf2、Brd2、Tspyl2、Eif4g2、Lcor、Isy1、Sf3a3、Msl2l1、Myo9a、 Papolg、Vasp、Pdcd7、Zbtb11、RGD1559904、Csnklg1、Leng8、Ddx10、Arhgef3、 Smg6、Zfat、Tm9sf3、Otud5和Chd6表达上调,在温阳消饮法代表方干预下表达下调；Ksr1、Slc25a27、Ptrf、Efemp1、RGD1563319、Cyb5r3、Gpr135、Polr3gl、 Kcna5、Ugp2、Acad11和Btbdll在造模条件下表达下调,用温阳消饮法代表方治疗后上调。 结论：前期研究发现,温阳消饮法有助于λ-角叉菜胶诱导的豚鼠胸腔积液的吸收。本实验表明,其机制可能与苓桂术甘汤合葶苈大枣泻肺汤给药调控多个生物学过程及信号通路有关。本研究从基因水平揭示了温阳消饮法的部分机制：①温阳消饮法可下调壁层胸膜炎症促进基因Mef2a、凋亡诱导基因Dyrk2和Tp53i11表达,改善炎症效应。②温阳消饮法可下调脏层胸膜炎症促进基因Rhoa、Figf、 Brd4、Crebbp和Brd2表达,上调炎症抑制基因Ksr1和Slc25a127表达,从而降低炎性损害。此外,Fgd4、Fh13和Nme7等基因的表达变化与本研究的关系有待进一步确认。
[Abstract]:Aim: to analyze the effect of Wenyang Xiaoyin method on pleural gene expression in pleural effusion rats.Methods Forty eight male Sprague-Dawley rats were randomly divided into group A (n = 40) and control group (n = 8) after anesthesia with pentobarbital sodium, 1% 位 -carrageenin solution was injected into the right pleural cavity for 24 hours.The chest X-ray films were observed by CR digital imaging system and the rats with obvious pleural effusion were screened out.The successful animals were randomly divided into Wenyang Xiaoyin group (n = 10), Wenyang Xiaoyin group (n = 10) and normal saline group (n = 9).The control group (n = 8) was perfused with normal saline once a day (5 ml / kg).After 4 days of continuous administration, animals were euthanized.The right visceral and parietal pleura of 3 animals in each group were collected and the expression of genes in pleural tissue was analyzed by Agilent single channel microarray.Results:Parietal pleural tissueThe differentially expressed genes in model group and blank control group, Wenyang Xiaoyin group and model group were involved in many biological processes and signaling pathways.Under the action of modeling factors, the expression of Fgd4Mef2aFhl3Dyrk2C24a6, Pbrm1, LOC498368, Tp53i11Rcan3Rcan3Rcan3Hpln4Abil and Hlx were up-regulated, and the expression of Nme7Pold2Klk1l and Fkbp5 were down-regulated under the condition of modeling, and up-regulated after administration of traditional Chinese medicine.Visceral pleural tissueThe differentially expressed genes in model group and blank control group, Wenyang Xiaoyin group and model group were involved in many biological processes and signaling pathways.鍦ㄩ，

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