Hernandes L, Zucoloto S, Alvares E P
Department of Histology and Embryology, Institute of Biomedical Sciences, University of São Paulo, SP, Brazil.
Cell Prolif. 2000 Jun;33(3):127-38. doi: 10.1046/j.1365-2184.2000.00172.x.
The effects of myenteric denervation on the cell kinetics of the intestinal epithelium of suckling and weanling rats were investigated. The myenteric plexus of an ileal segment was partially ablated by serosal application of benzalkonium chloride (BAC) in three groups of rats: those that underwent surgery at 13 days and were killed 15 (13/28-day-old) or 23 (13/36-day-old) days after treatment, and those that were operated at 21 days (21/36-day-old) and were killed 15 days after treatment. The extent of denervation was assessed in whole-mount preparations. The cell bodies of myenteric neurones were stained by NADH-diaphorase histochemical technique. Cell proliferation was estimated by the mitotic index (MI) and morphometric analysis of villus and crypt lengths using an image analysis system. Thickness of the muscle layers was also assessed by morphometry. Cell migration on the villi was estimated by the position of the leading labelled cell 24 h after tritiated thymidine injection. The number of neurones was reduced by around 80% in rats operated at 13 days, and reduced by 98% in those operated at 21 days. The thickness of the muscle layers was increased in all groups of treated animals. MI was significantly higher 15 days after BAC-treatment in the 13/28 group. Morphological changes in the intestinal mucosa were observed 15 days after BAC-treatment, when there was an increase in villus height (13/28 group) and crypt depth (13/28 and 21/36 groups). Cell migration rate was accelerated in the 21/36 group. No differences where found in the 13/36 group. These results show the strong effect of myenteric ablation on cell proliferation and migration in the ileal epithelium in the first 15 days of treatment in suckling and in weanling rats, and the subsequent recovery of intestinal mucosa homeostasis later on.
研究了肠肌层去神经支配对乳鼠和断奶幼鼠肠上皮细胞动力学的影响。三组大鼠通过在浆膜上应用苯扎氯铵(BAC)部分切除回肠段的肠肌丛:一组在13日龄时接受手术,在治疗后15天(13/28日龄)或23天(13/36日龄)处死;另一组在21日龄(21/36日龄)接受手术,在治疗后15天处死。在整装制片中评估去神经支配的程度。采用NADH - 黄递酶组织化学技术对肠肌神经元的细胞体进行染色。通过有丝分裂指数(MI)以及使用图像分析系统对绒毛和隐窝长度进行形态计量分析来估计细胞增殖。还通过形态计量学评估肌层厚度。通过注射氚标记胸腺嘧啶核苷24小时后领先标记细胞的位置来估计绒毛上的细胞迁移。13日龄手术的大鼠神经元数量减少约80%,21日龄手术的大鼠神经元数量减少98%。所有处理组动物的肌层厚度均增加。BAC处理后15天,13/28组的MI显著更高。BAC处理后15天观察到肠黏膜的形态学变化,此时绒毛高度增加(13/28组),隐窝深度增加(13/28组和21/36组)。21/36组的细胞迁移速率加快。13/36组未发现差异。这些结果表明,在乳鼠和断奶幼鼠治疗的前15天,肠肌层切除对回肠上皮细胞增殖和迁移有强烈影响,随后肠黏膜内环境稳定会逐渐恢复。