Effects of long-term ketamine administration on rat bladder protein levels: A proteomic investigation using two-dimensional difference gel electrophoresis system
Effects of long-term ketamine administration on rat bladder protein levels: A proteomic investigation using two-dimensional difference gel electrophoresis system
复制标题
长期服用氯胺酮对大鼠膀胱蛋白水平的影响：利用二维差异凝胶电泳系统进行蛋白质组学研究
DOI：
10.1111/iju.12100
复制
发表时间：
2013-10-01
影响因子：
2.6
通讯作者：
Wu, Peng
中科院分区：
文献类型：
--
作者：
Gu, Di;Huang, Jun;Wu, Peng
目的长期滥用氯胺酮可影响泌尿系统，导致间质性膀胱炎样综合征。然而，其发病机制尚不清楚。本研究采用双向差示凝胶电泳法结合基质辅助激光解吸电离光质谱技术，对氯胺酮相关性膀胱炎大鼠模型中潜在的疾病相关蛋白进行了研究。方法将大鼠随机分为对照组、生理盐水组、氯胺酮低剂量组和氯胺酮高剂量组，每组6只。试验组和对照组分别给予盐酸氯胺酮灌胃。生理盐水组每日灌胃生理盐水。治疗16周后，摘除膀胱，对生理盐水组和氯胺酮高剂量组的标本进行双向差示凝胶电泳法分析。用基质辅助激光解吸/电离光质谱鉴定差异表达斑点。结果高剂量氯胺酮处理组大鼠膀胱上皮细胞增生，炎性细胞浸润。双向差异凝胶电泳法显示，生理盐水组和氯胺酮高剂量组之间有30个基因表达发生了变化。在这些蛋白质中，两个上调的蛋白质点和两个下调的蛋白质点都被鉴定为平滑肌蛋白-22/转明胶。免疫组织化学染色和Western印迹分析显示，各组之间总转明胶的表达无明显差异。与生理盐水组比较，氯胺酮低、高剂量组大鼠膀胱壁磷酸化转明胶表达增加，非磷酸化转明胶表达降低。结论长期滥用氯胺酮可诱导膀胱壁转明胶磷酸化，这可能在氯胺酮相关性膀胱炎的发病机制中起重要作用。
ObjectivesLong-term ketamine abuse can affect the urinary system, resulting in interstitial cystitis-like syndrome. However, its pathogenesis remains unclear. In the present study, a proteomic approach of two-dimensional difference gel electrophoresis followed by matrix-assisted laser desorption/ionization time-of-light mass spectrometry was carried out to investigate the potential disease-associated proteins in a rat model of ketamine-associated cystitis.MethodsRats were randomly assigned to control, normal saline, low dose of ketamine (10mg/kg) and high-dose of ketamine (50mg/kg) groups with six rats in each group. The two experimental groups were given ketamine hydrochloride i.p. daily, whereas the normal saline group rats were treated with saline. After 16weeks of treatment, all bladders were excised, and samples from normal saline and high dose of ketamine groups were resolved in two-dimensional difference gel electrophoresis. Differentially expressed spots were excised and identified by matrix-assisted laser desorption/ionization time-of-light mass spectrometry. Phosphoprotein and non-phosphoprotein purification, histopathology, immunohistochemistry, and western blot were carried out in all groups.ResultsHistological study showed hyperplastic epithelium and inflammatory cells infiltration in the high dose of ketamine-treated rat bladders. Two-dimensional difference gel electrophoresis revealed 30 altered expressions between the normal saline and high dose of ketamine-treated group. Among these proteins, two upregulated and two downregulated protein spots were all identified as smooth muscle protein-22/transgelin. Immunohistochemical staining and western blot analysis showed that the expression of total transgelin had no significant difference between groups. However, the expression of phosphorylated transgelin in the low-dose and high dose of ketamine groups was increased, whereas the non-phosphorylated transgelin was decreased when compared with the normal saline group.ConclusionsLong-term ketamine abuse induces phosphorylation of transgelin in the bladder wall, and this might play an important role in the pathogenesis of ketamine-associated cystitis.