大鼠脊髓损伤后结肠功能变化与Cajal间质细胞的关系  

Relationship between colonic function change and interstitial cells of Cajal in rats with spinal cordinjury

作　　者：洪有建 江龙[1] 陈旺[1] 李陈成[1] 李森 刘媛 伍亚民 黄显凯[1] 

机构地区：[1]第三军医大学大坪医院野战外科研究所创伤外科,重庆400042 [2]第三研究室,创伤、烧伤与复合伤国家重点实验室

出　　处：《中华创伤杂志》2016年第11期1028-1033,共6页Chinese Journal of Trauma

基　　金：国家自然科学基金（30772299）

摘　　要：目的探讨大鼠脊髓损伤后结肠功能的改变与Cajal间质细胞（ICC）变化的关系，进一步分析脊髓损伤后肠功能障碍的形成机制。方法采用改良Allen装置以10g×25cm强度打击大鼠T10硬膜囊，建立脊髓损伤模型。选择100只成年sD雌性大鼠按随机数字表法分为正常组（20只）、假手术组（20只）和损伤组（60只）。各组于伤后1，2，4周取近端结肠组织，行HE染色观察形态学改变；记录各组近端结肠的生物电改变；免疫荧光双标染色检测各组结肠肌层ICC阳性细胞数和神经元吸光度（A）值的变化；Westernblot检测伤后1，2，4周时各组结肠组织ICC特异蛋白c—kit的表达变化。结果HE染色可见正常组和假手术组近端结肠绒毛上皮细胞排列整齐，损伤组绒毛上皮细胞倒伏、脱落，黏膜层及肌层厚度增加。正常组和假手术组生物电活动较规律，损伤组近端结肠生物电紊乱，振幅变异较大，且波形较不稳定。免疫荧光染色显示损伤组近端结肠肌内ICC阳性细胞数1周为57．384-4．69，2周为32．26±3．62，4周为25．07±3．05，均明显少于正常组和假手术组（P〈0．05）。2周时损伤组结肠神经元A值为71．46±4．69，均小于正常组的114．30±24．44和假手术组的108．11±26．46（P〈0．05）。Westernbla显示损伤组C—kit蛋白表达1周为0．7454-0．017，2周为0．492±0．480，4周为0．409±0．032，较正常组和假手术组明显减少（P〈0．05）。结论大鼠脊髓损伤后结肠功能明显改变，可能与结肠肌内ICC数量减少和C—kit的表达降低有关。Objective To investigate the correlation of colonic function change with interstitial cells of Cajal （ICC） in rats with spinal cord injury and further evaluate the mechanism of intestinal dysfunction after spinal cord injury. Methods Using the modified Allen＇ s device, experimental spinal cord injury was induced by dropping a 10 g weight from the height of 25 cm on the T10 segment. A total of 10 male adult SD rats were divided into normal group （n = 20 ）, sham-operated group （n = 20 ） and injury group （ n = 60） , according to the random number table. At postoperative 1, 2 and 4 weeks, proximal colon tissues in each group were collected for morphological study using HE staining, bioelectricity study, ICC count and neuron absorbance value using double-labeling immunofluorescencc staining, and expression of c-kit protein in ICC using Western blot method. Results Proximal colon epithelial cells arranged neatly in normal and sham-operated groups, while the lodging and shedding of villus epithelial cells and increased thickness of mucous layer and muscular layer were seen in injury group. Bioelectricityin injury group disordered with large amplitude variation and unstable waveform, compared to the regular biological activity in normal and sham-operated groups. Intramuscular ICC cell count in injury group was 57.38 ± 4.69 at 1 week, 32.26 ± 3.62 at 2 weeks and 25.07 ±3.05 at 4 weeks, significantly less than that in normal and sham-operated groups（P 〈 0.05 ）. At 2 weeks, mean absorbance value of the neuron in injury group （71.46 ± 4.69） was less than that in normal and the sham-operated groups （ 114.30 ±24.44, 108.11 ±26.46） （ P 〈 0.05 ）. Expression of c-kit protein in ICC ceils was 0. 745 ± 0. 017 at 1 week, 0.492 ± 0.480 at 2 weeks and 0.409 ±0. 032 at 4 weeks, significantly less than that in normal and sham-operated groups （P 〈0.05）. Conclusion Colonic function in rats presents significant changes after spinal cord injury, and the changes may relate to the decrease

关 键 词：肠 CAJAL间质细胞 脊髓损伤 

分 类 号：R651.2[医药卫生—外科学]