白头翁汤对内毒素诱导血管内皮细胞基因表达谱的影响

doi:10.11843/j.issn.0366-6964.2013.10.023

畜牧兽医学报 ›› 2013, Vol. 44 ›› Issue (10): 1675-1684.doi: 10.11843/j.issn.0366-6964.2013.10.023

白头翁汤对内毒素诱导血管内皮细胞基因表达谱的影响

胡屹屹¹，穆祥²，胡元亮³

（1. 江苏省农业科学院兽医研究所·农业部兽用生物制品工程技术重点实验室·国家兽用生物制品工程技术研究中心，南京210014；2. 北京农学院兽医学中医药北京市重点实验室，北京 102206；3. 南京农业大学动物医学院，南京 210095）

收稿日期:2013-04-01 出版日期:2013-10-23 发布日期:2013-10-28
作者简介:胡屹屹（1981-），男，南京市雨花区人，助理研究员，博士，主要从事中兽医学和动物药理学研究，E-mail: huyiyi6073@sina.com
基金资助:
江苏省农业科技自主创新基金［cx(13)5030］

Effects of Pulsatillae Decoction on Gene Expression of LPS-induced Endothelial Cell

HU Yi-yi¹, MU Xiang², HU Yuan-liang³

（1. Key Laboratory of Veterinary Biological Engineering and Technology of Ministry of Agriculture·National Center for Engineering Research of Veterinary Bio-products, Institute of Veterinary Medicine, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China;2. Beijing Key Laboratory of Traditional Chinese Veterinary Medicine, Beijing University of Agriculture, Beijing 102206, China; 3. College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, China）

Received:2013-04-01 Online:2013-10-23 Published:2013-10-28

摘要/Abstract

摘要：

拟探讨中药白头翁汤方剂（PD）对细菌内毒素（LPS）损伤的治疗作用。将内皮细胞培养至单层融合状态，加入LPS和白头翁汤作用12 h后，提取3个处理组的总RNA。采用Affymetrix Rat Genome 230 2.0 Array芯片检测，读取各组基因表达的变化情况并进行生物学分析。选取5个与炎症反应相关的差异表达基因，采用嵌合荧光法进行Real-time RT-PCR反应。结果显示：与空白组相比，LPS组产生69个差异表达基因，其中上调基因36个，下调基因33个；与LPS组相比，LPS-PD组产生578个差异表达基因，其中上调基因566个，下调基因12个；与空白组相比，LPS-PD组产生122个差异表达基因，其中上调基因93个，下调基因29个。5个目标基因Il1α、Il6、Edn1、Tnfsf11和Ptges的Real-time RT-PCR检测结果依次为9.78、1.76、1.43、2.98和2.12，对应的基因芯片检测结果分别是10.57、1.53、1.59、3.16和1.71，两者变化趋势相符。白头翁汤抗内毒素的药理机制可能在于减少炎症因子的蛋白表达，抑制凋亡基因的启动，阻碍内毒素信号传导与受体激活，增加机体能量代谢和蛋白质合成。

关键词: 白头翁汤, 内毒素, 基因芯片, 炎症

Abstract:

A high-throughput and efficient Affymetrix rat genome array was used to investigate the pharmacological mechanism of the traditional Chinese medicine, Pulsatillae Decoction (PD), used for the treatment of diseases induced by lipopolysaccharide (LPS). Endothelial cell was challenged with 1 μg·mL^－1 LPS for 3 h, then treated with PD at a concentration of 1 mg·mL^-1 for 12 h. Total RNA of each treated group was extracted from cultured endothelial cell for detection by the Affymetrix Rat Genome 230 2.0 Array. The samples from LPS group and blank control group were subjected to Real-time RT-PCR, using the chimeric fluorescence method. The results showed that in LPS group, 36 genes were upregulated and 33 genes were downregulated in comparison with Blank group; In LPS-PD group, 566 genes were upregulated and 12 genes were downregulated in comparison with LPS group, and 93 genes were upregulated and 29 genes were downregulated in comparison with Blank group. The RT-PCR ratios for the target genes Il1α, Il6, Edn1, Tnfsf11 and Ptges were 9.78, 1.76, 1.43, 2.98 and 2.12 respectively, which were very similar to the ratios determined by the gene chips (10.57, 1.53, 1.59, 3.16 and 1.71, respectively). These results indicate that PD could simultaneously reduce the translated level of inflammatory factors, suppress the transcription of apoptosis genes, accelerate the protein synthesis of organism and enhance the energy metabolism of cells, thus play the united antiendotoxin action.

Key words: Pulsatillae Decoction, LPS, gene chip, inflammation

中图分类号:

S853.9

胡屹屹，穆祥，胡元亮. 白头翁汤对内毒素诱导血管内皮细胞基因表达谱的影响[J]. 畜牧兽医学报, 2013, 44(10): 1675-1684.

HU Yi-yi, MU Xiang, HU Yuan-liang. Effects of Pulsatillae Decoction on Gene Expression of LPS-induced Endothelial Cell[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2013, 44(10): 1675-1684.

参考文献

［1］MOTAWI T K，DARWISH H A，ABD E L，et al. Effects of caffeic acid phenethyl ester on endotoxin-induced cardiac stress in rats: A possible mechanism of protection ［J］. J Biochem Mol Toxicol，2011，25(2):84-94.

［2］HAN J H，MCKENZIE H C，MCCUTCHEON L J，et al. Glucose and insulin dynamics associated with continuous rate infusion of dextrose solution or dextrose solution and insulin in healthy and endotoxin-exposed horses ［J］. Am J Vet Res，2011，72(4):522-529.

［3］ROH E，LEE H S，KWAK J A， et al. MD-2 as the target of nonlipid chalcone in the inhibition of endotoxin LPS-induced TLR4 activity ［J］. J Infect Dis，2011，203(7):1012-1020.

［4］陆树文，刘红菊，赵伟，等.白头翁汤治疗炎症性肠病的分子机制研究［J］.中国应用生理学杂志，2011，27(1)：106-109.

［5］王瑞锋，王雪.白头翁汤的药理作用与临床应用［J］.中医学报，2010，25(147)：270-271.

［6］LEONG H X，SIMKEVICH C，LESIEURBROOKS A，et al. Short-term arginine deprivation results in large-scale modulation of hepatic gene expression in both normal and tumor cells: microarray bioinformatic analysis ［J］. Nutr Metab (Lond)，2006，3: 37.

［7］BUETTNER R，PARHOFER K G，WOENCKHAUS M，et al. Defining high-fat-diet rat models: metabolic and molecular effects of different fat types ［J］. J Mol Endocrino，2006，36(3):485-501.

［8］YOSHIYUKI T，MITSUO T，HIROKI W，et al. Gene expression profiles of hepatic cell-type specific marker genes in progression of liver fibrosis ［J］. World J Gastroenterol，2006，12(40):6473-6499.

［9］EISEN M B，SPELLMAN P T，BROWN P O，et al. Cluster analysis and display of genome-wide expression patterns ［J］. Proc Natl Acad Sci，1998，95(25):14863-14868.

［10］LO Y C，TSAI P L，HUANG Y B，et al. San-Huang-Xie-Xin-Tang reduces lipopolysaccharides-induced hypotension and inflammatory mediators ［J］. J Ethnopharmacol，2005，96(1-2):99-106.

［11］TSENG S H，LEE H H，CHEN L G，et al. Effects of three purgative decoctions on inflammatory mediators ［J］. J Ethnopharmacol，2006，105(1-2):118-124.

［12］SUGANUMA T，IRIE K，FUJII E，et al. Effect of heat stress on lipopolysaccharide-induced vascular permeability change in mice ［J］. J Pharmacol Exp Ther，2002，303(2):656-663.

［13］JO S K，CHA D R，CHO W Y，et al. Inflammatory cytokines and lipopolysaccharide induce Fas-mediated apoptosis in renal tubular cells ［J］. Nephron，2002，91(3):406-415.

［14］BAMFORD S，RYLEY H，JACKSON S K. Highly purified lipopolysaccharides from Burkholderia cepacia complex clinical isolates induce inflammatory cytokine responses via TLR4-mediated MAPK signaling pathways and activation of NF kappaB ［J］. Cell Microbiol，2007，9(2):532-543.

［15］OHTA K，KIKUCHI T，MIYAHARA T，et al. DNA microarray analysis of gene expression in iris and ciliary body of rat eyes with endotoxin-induced uveitis ［J］. Exp Eye Res，2005，80(3):401-412.

［16］ORELLANA R A，O’CONNOR P M，NQUVEN H V，et al. Endotoxemia reduces skeletal muscle protein synthesis in neonates ［J］. Am J Physiol Endocrinol Metab，2002，283(5):E909-916.

［17］COIMBRA R，LOOMIS W，MELBOSTAD H，et al. LPS-stimulated PMN activation and proinflammatory mediator synthesis is downregulated by phosphodiesterase inhibition: role of pentoxifylline ［J］. J Trauma，2004，57(6):1157-1163.

［18］DELPECH M. DNA chips ［M］. Ann Biol Clin(Paris)，2000，58(1):29-38.

［19］DEBOUCK C，GOODFELLLOW P N. DNA microarrays in drug disease and development ［J］. Nat Genet，1999，21(1 Suppl):S48-50.

［20］LU A，TANG Y，RAN R，et al. Genomics of the periinfarction cortex after focal cerebral ischemia ［J］. J Cereb Blood Flow Metab，2003，23:786-810.

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白头翁汤对内毒素诱导血管内皮细胞基因表达谱的影响

Effects of Pulsatillae Decoction on Gene Expression of LPS-induced Endothelial Cell

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