肠内生态免疫营养对创伤后大鼠肠道免疫功能及损伤修复信号通路Hedgehog蛋白表达的影响

Effect of enteral ecological immune nutrition on intestinal immune function and damage repair signal pathway Hedgehog protein expression in rats after trauma

目的 观察肠内生态免疫营养制剂对创伤后大鼠肠道免疫功能及损伤修复信号通路Hedgehog蛋白的影响.方法 雄性清洁级Wistar大鼠40只采用小肠损伤法制作肠道损伤模型,并采用随机数字表法分为普通营养组（对照组,20只）和肠内生态营养组（实验组,20只）.对照组大鼠给予普通饲料灌胃,实验组大鼠给予瑞能＋金双歧灌胃（每只每天金双歧1×10CFU）.造模成功后12h开始给予752.76kJ/（kg·d）的标准等氮热量灌胃4次/日.灌胃1周后处死大鼠并切取两组大鼠小肠,进行苏木素-伊红（HE）染色观察小肠黏膜形态学改变：包括绒毛高度、肠腺隐窝深度;应用免疫组织化学技术检测小肠组织中CD3＋、CD4＋、CD8＋表达水平有无差异,采用Western blot法检测两组大鼠小肠黏膜Hedgehog蛋白表达的差异.结果 两组大鼠体重先降低后升高,从术后第5、6、7天实验组大鼠体重分别为（127.1±5.0）、（130.2±4.6）、（132.5±4.1）g,显著高于对照组的（123.2±4.2）、（125.8±3.9）、（128.4±5.2）g,差异有统计学意义（t5d=2.313,P5d=0.028;t6d=2.826,P6d=0.009;t7d=2.335,P7d=0.027）;镜下实验组大鼠小肠绒毛高度为（230.2±21.4）μm,小肠腺隐窝深度为（104.3±10.2）μm,黏膜厚度为（354.1±40.63）μm,均显著高于对照组 （P=0.000）;实验组大鼠小肠CD3＋、CD4＋及CD8＋阳性淋巴细胞数百分比分别为（34.2±4.9）%、（26.8±4.0）%和（22.3±3.4）%,实验组显著高于对照组;实验组大鼠小肠黏膜组织中Hedgehog蛋白相对表达量为0.14±0.03,显著高于对照组（0.05±0.02）,差异有统计学意义（t=11.160,P=0.000）.结论 肠内生态免疫营养可增强Wistar大鼠肠道免疫功能,促进大鼠小肠黏膜损伤修复,提高小肠中Hedgehog蛋白表达水平.

Objective To evaluate the effects of enteral ecological immune nutrition on intestinal immune function and damage repair signal pathway Hedgehog protein expression in rats after trauma.Methods Forty healthy male Wistar rats were made of small intestine trauma model and randomly divided into experimental and control groups with each 20 rats by random number table.The rats in control group were given regular nutrition forage and Wistar rats in the experiment group were given live combined bifidobacterium and lactobacillus tablets with 1×10CFU per-day.At 12 h after operation, the rats were given 180 kcal/（kg·d） standard and other nitrogen gavage, 4 times every day.One week after nutrition gavage, the rats in the two groups were killed and the morphological changes of small intestinal mucosa, the CD3＋, CD4＋, CD8＋ positive lymph cells and Hedgehog protein expression were compared between the two groups by hematoxylin and eosin （HE） stain, immunohistochemical technique and Western blotting.Results The body weight of the two groups were firstly decreased and then increased.The body weight of experiment group for day 5, 6 and 7 were （127.1±5.0）, （130.2±4.6） and （132.5±4.1） g respectively which significant higher than control group （123.2±4.2）, （125.8±3.9） and （128.4±5.2） g with statistical difference （t5 d=2.313, P5 d=0.028;t6 d=2.826, P6 d=0.009;t7 d=2.335, P7 d=0.027）.The villus height, intestinal gland crypt depth and mucosal thickness （230.2±21.4）, （104.3±10.2） and （354.1±40.63） μm in the experimental group, which were significantly inceased as compared with those in the control group （P=0.000）.The CD3＋, CD4＋ and CD8＋ positive percentage were （34.2±4.9）%, （26.8±4.0）% and （22.3±3.4）% in the experimental group, which was significantly higher than in the control group.The Hedgehog protein expression in the control group and experimental group was 0.05±0.02 and 0.14±0.03 respectively with the difference being statistically significant （P=0.000）.Conclusion Enteral ecological immune nutrition can enhance the immune function of Wistar rats, promote the repair of intestinal mucosal damage in rats, and improve the expression level of Hedgehog protein in the small intestine.