Gene expression profiling and endothelin in acute experimental pancreatitis
Gene expression profiling and endothelin in acute experimental pancreatitis
复制标题
DOI：
10.3748/wjg.v18.i32.4257
复制
发表时间：
2012-08-28
影响因子：
4.3
通讯作者：
Westlund, Karin N.
中科院分区：
文献类型：
--
作者：
Oz, Helieh S.;Lu, Ying;Westlund, Karin N.
目的：分析实验性胰腺炎的基因表达谱，并提供内皮素-1拮抗剂候选基因的功能逆转。方法：二氯化二丁基锡（DBTC）是一种用于聚碳酸酯稳定剂/催化剂、农业杀菌剂、油漆和织物防污剂的化学品。DBTC通过产生活性氧诱导急性胰腺炎发作。lewis近交系大鼠接受单次静脉注射DBTC或载药。在炎症高峰期采集脊髓和背根神经节（DRG），并使用偏向于大鼠脑特异性基因的cDNA微阵列进行转录分析。在第二项研究中，用内皮素（Er）受体拮抗剂[ET-A （BQ123）和ET-B BQ788]治疗dbtc诱导的胰腺炎动物组。测量自发性疼痛相关的机械和热超敏反应。采用抗er - a和ET-B抗体对胰腺组织切片和T10-12脊柱节段DRG进行免疫组化分析。结果：动物发生急性胰腺炎症，持续7-10 d，病理检查证实（实质水肿、胰腺结构和胰岛丧失、炎症细胞、中性粒细胞和单核细胞浸润、腺泡细胞变性、空泡化和坏死）和疼痛相关行为（皮肤继发性机械和热过敏）。与对照组相比，胰腺炎动物脊髓中的基因表达谱有所不同。260多个上调和60多个下调的独特基因可分为8个功能基因家族：循环/急性期/免疫调节；细胞外基质;结构;频道/受体/转运体;信号转导;转录/翻译相关;抗氧化剂/监护人/热休克;胰酶和其他酶。ET-1是52个候选基因之一，在胰腺炎症和内脏疼痛相关行为的动物中上调幅度超过2倍。Er-A （BQ123）和ET-B （BQ-788）拮抗剂对胰腺炎动物炎症性疼痛相关的机械和热过敏行为有显著的保护作用（P < 0.05）。裸场自发行为活动（BQ123、BQ788用药后基线、第6天和30 min）呈现总体稳定的活动水平，表明药物对正常探索行为没有不良影响，但在最高剂量（300 μ mol/L）下，药物有活动时间减少、休息时间增加的趋势。免疫细胞化学定位显示胰腺炎动物DRG中ET-A和ET-B受体表达增加。内皮素受体定位结合神经元标记物NeuN和胶质标记物胶质原纤维酸性蛋白双染色。ET-A在幼稚动物DRG神经元的细胞体和偶有细胞核中表达。然而，胰腺炎动物的所有大小神经元中ET-A受体的表型表达均显著增加。同样，ET-B受体定位于神经元和卫星胶质，以及通过DRG的轴突周围的雪旺细胞胶质髓鞘。结论：内皮素受体拮抗剂对炎性疼痛反应有保护作用，但不干扰正常的探索行为。候选基因可以作为未来诊断和/或靶向基因治疗的生物标志物。(c) 2012年白石登。版权所有。
AIM: To analyze gene expression profiles in an experimental pancreatitis and provide functional reversal of hypersensitivity with candidate gene endothelin-1 antagonists.METHODS: Dibutyltin dichloride (DBTC) is a chemical used as a polyvinyl carbonate stabilizer/catalyzer, biocide in agriculture, antifouling agent in paint and fabric. DBTC induces an acute pancreatitis flare through generation of reactive oxygen species. Lewis-inbred rats received a single i.v. injection with either DBTC or vehicle. Spinal cord and dorsal root ganglia (DRG) were taken at the peak of inflammation and processed for transcriptional profiling with a cDNA microarray biased for rat brain-specific genes. In a second study, groups of animals with DBTC-induced pancreatitis were treated with endothelin (Er) receptor antagonists [ET-A (BQ123) and ET-B BQ788)]. Spontaneous pain related mechanical and thermal hypersensitivity were measured. Immunohistochemical analysis was performed using anti-Er-A and ET-B antibodies on sections from pancreatic tissues and DRG of the T10-12 spinal segments.RESULTS: Animals developed acute pancreatic inflammation persisting 7-10 d as confirmed by pathological studies (edema in parenchyma, loss of pancreatic architecture and islets, infiltration of inflammatory cells, neutrophil and mononuclear cells, degeneration, vacuolization and necrosis of acinar cells) and the pain-related behaviors (cutaneous secondary mechanical and thermal hypersensitivity). Gene expression profile was different in the spinal cord from animals with pancreatitis compared to the vehicle control group. Over 260 up-regulated and 60 down-regulated unique genes could be classified into 8 functional gene families: circulatory/acute phase/immunomodulatory; extracellular matrix; structural; channel/receptor/transporter; signaling transduction; transcription/translation-related; antioxidants/chaperones/heat shock; pancreatic and other enzymes. ET-1 was among the 52 candidate genes up-regulated greater than 2-fold in animals with pancreatic inflammation and visceral pain-related behavior. Treatments with the Er-A (BQ123) and ET-B (BQ-788) antagonists revealed significant protection against inflammatory pain related mechanical and thermal hypersensitivity behaviors in animals with pancreatitis (P < 0.05). Open field spontaneous behavioral activity (at baseline, day 6 and 30 min after drug treatments (BQ123, BQ788) showed overall stable activity levels indicating that the drugs produced no undesirable effects on normal exploratory behaviors, except for a trend toward reduction of the active time and increase in resting time at the highest dose (300 mu mol/L). Immunocytochemical localization revealed that expression of ET-A and ET-B receptors increased in DRG from animals with pancreatitis. Endothelin receptor localization was combined in dual staining with neuronal marker NeuN, and glia marker, glial fibrillary acidic protein. ET-A was expressed in the cell bodies and occasional nuclei of DRG neurons in naive animals. However, phenotypic expression of ET-A receptor was greatly increased in neurons of all sizes in animals with pancreatitis. Similarly, ET-B receptor was localized in neurons and in the satellite glia, as well as in the Schwann cell glial myelin sheaths surrounding the axons passing through the DRG.CONCLUSION: Endothelin-receptor antagonists protect against inflammatory pain responses without interfering with normal exploratory behaviors. Candidate genes can serve as future biomarkers for diagnosis and/or targeted gene therapy. (c) 2012 Baishideng. All rights reserved.