Lantéri-Minet M, Bon K, de Pommery J, Michiels J F, Menétrey D
Institut National de la Santé et de la Recherche Médicale Unité 161, Paris, France.
Exp Brain Res. 1995;105(2):220-32. doi: 10.1007/BF00240958.
The evoked expression of the immediate early gene (IEG)-encoded proteins c-Fos and Krox-24 was used to monitor spinal visceronociceptive processing that results from cyclophosphamide cystitis in behaving rats. Animals received a single dose of 100 mg/kg i.p. of cyclophosphamide and survived for 30 min to 5 h. Longer survival times were not considered because of ethical considerations. Cyclophosphamide-injected animals developed characteristic behavioral signs in parallel with development of bladder lesions and spinal evoked expression of IEG-encoded proteins. Histological examination of the urinary bladder was used to evaluate the degree of cystitis and as a criterion for selection of groups of animals to be quantitatively analyzed. Controls consisted of freely behaving animals including control (un-injected), sham (saline-injected) or diuretic (furosemide-injected) animals. Behavioral modifications consisted of lacrimation, piloerection, assumption of a peculiar "rounded-back" posture, which was accompanied by head immobility and various brief "crises" (tail hyperextension, abdominal retractions, licking of the lower abdomen, backward withdrawal movements). Abnormal behaviors, which first appeared (lacrimation, piloerection) at the end of postinjection hour 1, progressively increased in severity (rounded-back posture) over the following 90 min to reach a plateau at about postinjection hour 2; the rounded-back posture was maintained up to time of death. Histological modifications of bladder tissue were assessed using a 4-grade scale in a blind setting. The 1st grade consisted of control or sham animals with no bladder lesion; 2nd grade, animals with simple chorionic edema; 3rd grade, animals with chorionic edema associated with mucosal abrasion, fibrin deposit, and onset of polymorphonuclear leukocyte infiltration; 4th grade, animals with complete cystitis corresponding to an increase in severity and spread of all the signs of cystitis described above plus petechial hemorrhage. Simple chorionic edema was observed from 30 min to 3 h postinjection, but with a progressive increase in severity over time. Edema accompanied by epithelial abrasion was observed for animals that survived 3-4 h postinjection; complete inflammation was observed in animals that survived 4-5 h postinjection. The study of c-Fos- and Krox-24-encoded protein expression demonstrated that few lumbosacral spinal areas were specifically involved in the processing of visceral inputs in response to bladder stimulation. These areas were the parasympathetic column (SPN), the dorsal gray commissure (DGC as the caudal extent of lamina X), and superficial layers of the dorsal horn.(ABSTRACT TRUNCATED AT 400 WORDS)
立即早期基因（IEG）编码蛋白c-Fos和Krox-24的诱发性表达被用于监测行为学实验大鼠中由环磷酰胺膀胱炎引起的脊髓内脏伤害性感受处理过程。动物腹腔注射100mg/kg的环磷酰胺单剂量，存活30分钟至5小时。出于伦理考虑，未考虑更长的存活时间。注射环磷酰胺的动物出现了与膀胱损伤发展以及IEG编码蛋白的脊髓诱发性表达并行的特征性行为体征。对膀胱进行组织学检查以评估膀胱炎的程度，并作为选择用于定量分析的动物组的标准。对照组包括自由活动的动物，即对照（未注射）、假手术（注射生理盐水）或利尿（注射速尿）动物。行为改变包括流泪、竖毛、呈现特殊的“圆背”姿势，同时伴有头部不动和各种短暂的“发作”（尾巴过度伸展、腹部收缩、舔舐下腹部、向后退缩动作）。异常行为在注射后第1小时末首次出现（流泪、竖毛），在接下来的90分钟内严重程度逐渐增加（圆背姿势），在注射后约第2小时达到平台期；圆背姿势一直维持到死亡。在盲法设置下，使用4级量表评估膀胱组织的组织学改变。第1级包括无膀胱损伤的对照或假手术动物；第2级，有单纯绒毛膜水肿的动物；第3级，有与粘膜擦伤、纤维蛋白沉积和多形核白细胞浸润开始相关的绒毛膜水肿的动物；第4级，有完全膀胱炎的动物，对应于上述膀胱炎所有体征的严重程度增加和扩散，加上瘀点出血。在注射后30分钟至3小时观察到单纯绒毛膜水肿，但随着时间推移严重程度逐渐增加。在注射后存活3 - 4小时的动物中观察到伴有上皮擦伤的水肿；在注射后存活4 - 5小时的动物中观察到完全炎症。对c-Fos和Krox-24编码蛋白表达的研究表明，很少有腰骶部脊髓区域特异性参与对膀胱刺激的内脏传入处理。这些区域是副交感神经柱（SPN）、背侧灰质连合（作为X层尾端范围的DGC）和背角浅层。（摘要截断于400字）
