温阳消饮法对胸腔积液模型大鼠肾、空肠组织差异基因表达影响的实验研究

发布时间：2019-01-02 14:29

【摘要】：目的：利用基因芯片技术分析温阳消饮法调控胸腔积液模型大鼠基因表达的作用机制。方法：40只清洁级Wistar成年大鼠适应性饲养7天后,随机分为温阳消饮组(W)、去桂消饮组(Q)、模型对照组(M)、空白对照组(K),每组10只。除空白对照组外,其余3组以1%λ-角叉菜胶胸腔注射建立胸腔积液(类似悬饮)大鼠模型,造模24h后拍X线片对各组大鼠进行病理评价。于造模后2h各组以相应药物灌胃,每天1次,连续5天。灌胃结束后大鼠进行安乐死。每组取3只大鼠的肾髓质和空肠组织,采用基因芯片技术进行基因表达谱检测,筛选出差异表达基因,并进行Pathway分析。结果：1.肾髓质组织基因表达谱检测温阳消饮组：温阳消饮组与模型对照组比较和模型对照组与空白对照组比较共同表达的通路为：神经活性配体受体相互作用通路、系统性红斑狼疮、肾素血管紧张素系统,共同表达基因为:LOC317471、Olr462、Mcpt8I3、Olr718、Mcpt8I2、 Cd80、Ttn。去桂消饮组：去桂消饮组与模型对照组比较和模型对照组与空白对照组比较共同表达的通路为：神经活性配体受体相互作用通路,共同表达的基因为Cep295nl、Mcpt8I3、Fam64a。2.空肠组织基因表达谱检测温阳消饮组：温阳消饮组与模型对照组比较和模型对照组与空白对照组比较共同表达的通路为：药物代谢细胞色素P450通路、卟啉和叶绿素代谢通路、淀粉和蔗糖代谢通路、甾类激素生物合成通路、嗅觉传导通路、药物其他酶代谢通路、细胞色素P450外源性化学物质通路、戊糖和葡萄糖醛酸相互转换通路、视黄酵代谢通路,共同表达的基因为：Cacnb4、Gpr27、Ugt2b、Spock3、Olr113、 Olr1472、Ugt2b7、Opn5、RGD1564865、Ugt2b17、Ugt2b37、Slc5a5、Slc4a9、 Ccdc79、Anks1b。去桂消饮组：去桂消饮组与模型对照组比较和模型对照组与空白对照组比较共同表达的通路为：药物代谢细胞色素P450通路,共同表达的基因为：Kcnq2、 LOC102548590、Cyp2b12、Cyp2b21、Aldhla1、Cyp2d3、Arnt2。结论：1.温阳消饮法可以减少λ-角叉菜胶诱导的胸腔积液模型大鼠的胸腔积液量。2.利用基因芯片技术对胸腔积液模型大鼠肾髓质进行差异基因分析,发现温阳消饮方与去桂消饮方可能是通过上调Mcpt8I3基因调控神经活性配体受体相互作用通路,参与炎症反应。3.温阳消饮组与去桂消饮组在肾髓质组织的pathway分析中存在差异,温阳消饮方可能是通过上调Mcpt8I3基因调控肾素血管紧张素系统通路达到利尿消饮的目的。4.利用基因芯片技术对胸腔积液模型大鼠空肠组织进行pathway分析,去桂消饮组未发现有与利尿作用相关的通路,而温阳消饮方可能通过调控甾类激素生物合成通路,抑制醛固酮的合成来达到利尿的作用。5.本研究中未发现cAMP-PKA/PKC及VWP通路,提示温阳消饮方可能不是通过该通道发挥利尿或抑制尿液重吸收作用的。
[Abstract]:Aim: to analyze the mechanism of Wenyang Xiaoyin method in regulating gene expression in rats with pleural effusion. Methods: 40 clean grade Wistar adult rats were fed for 7 days, and were randomly divided into Wanyang Xiaoyin group, (W), group, (Q), model control group, (M), blank control group, (K), group, 10 rats in each group. In addition to the blank control group, the other three groups were injected with 1% 位 -carrageenin into the pleural cavity to establish the model of pleural effusion (similar to suspension drink). Two hours after modeling, each group was gavaged with corresponding drugs once a day for 5 days. The rats were euthanized after gavage. The kidney medulla and jejunum of 3 rats in each group were collected and the differentially expressed genes were screened by gene chip technique and analyzed by Pathway. Results: 1. The gene expression profile of renal medulla tissue in Wenyang Xiaoyin group: compared with the model control group and the model control group compared with the blank control group: the neuroactive ligand receptor interaction pathway, systemic lupus erythematosus. Renin angiotensin system, co-expressed by LOC317471,Olr462,Mcpt8I3,Olr718,Mcpt8I2, Cd80,Ttn. Compared with the model control group and the model control group, the co-expression pathway of the Deiguixiao Yin group and the blank control group was: the neuroactive ligand receptor interaction pathway, and the co-expressed gene was Cep295nl,Mcpt8I3,Fam64a.2.. Detection of gene expression profile in jejunum tissue: the pathways of gene expression in Wenyang Xiaoyin group and model control group were as follows: drug metabolism cytochrome P450 pathway, porphyrin and chlorophyll metabolism pathway, compared with model control group and model control group. Starch and sucrose metabolism pathway, steroid hormone biosynthesis pathway, olfactory pathway, drug other enzyme metabolism pathway, cytochrome P450 exogenous chemical pathway, pentose and glucuronic acid interconversion pathway, retinozyme metabolism pathway, The co-expressed genes are: Cacnb4,Gpr27,Ugt2b,Spock3,Olr113, Olr1472,Ugt2b7,Opn5,RGD1564865,Ugt2b17,Ugt2b37,Slc5a5,Slc4a9, Ccdc79,Anks1b. Compared with the model control group and the model control group, the drug metabolism cytochrome P450 pathway and the co-expressed gene Kcnq2, LOC102548590,Cyp2b12,Cyp2b21,Aldhla1,Cyp2d3,Arnt2. were found in the Deiguixiao decoction group and the model control group and the blank control group respectively. Conclusion: 1. Wenyang Xiaoyin method can reduce the amount of pleural effusion induced by 位-carrageenin in rats. 2. The differential gene analysis of renal medulla in rats with pleural effusion was carried out by using gene chip technique. It was found that Wenyang Xiaoyin recipe and Deiguixiaoyin prescription might regulate the neuroactive ligand receptor interaction pathway by up-regulating Mcpt8I3 gene. Participation in inflammatory response. 3. There were differences in pathway analysis of renal medulla between Wenyangxiaoyin group and Danguixiaoyin group. Wenyangxiaoyin prescription could achieve diuretic Xiaoyin by up-regulating the renin angiotensin system pathway by up-regulating Mcpt8I3 gene. 4. The pathway analysis of jejunum tissue of rats with pleural effusion model was carried out by using gene chip technology. There was no diuretic pathway in the group of deguixiao drink, while Wenyang Xiaoyin prescription could regulate steroid hormone biosynthesis pathway. Inhibition of aldosterone synthesis to achieve diuretic effect. 5. The cAMP-PKA/PKC and VWP pathways were not found in this study, suggesting that WYXF may not play diuretic or inhibit urinary reabsorption through this channel.
【学位授予单位】：成都中医药大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R285.5;R-332

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