摘要
目的:观察糖尿病胃肠功能紊乱模型大鼠结肠组织内P物质的分布和含量变化,分析P物质在糖尿病胃肠功能紊乱发病中的作用。方法:实验于2005-03/08在南京医科大学动物中心和南京医科大学第一附属医院消化内科实验室完成。取30只SD大鼠随机分为正常对照组(n=10)和糖尿病组(n=20)。糖尿病组大鼠单剂量腹腔注射链脲佐菌素80mg/kg建立糖尿病大鼠模型,而正常对照组注射等量枸橼酸缓冲液,72h后尾静脉采血测血糖,血糖≥16.7mmol/L为成功模型。两组大鼠饲养6周后脱臼处死,计算无张力下胃肠推进率,并应用免疫组化测定P物质在大鼠结肠组织内的表达,以及应用反转录-聚合酶链反应法测定大鼠结肠组织内P物质mRNA的表达。结果:进入结果分析大鼠20只,正常对照组10只,糖尿病组10只。①P物质在糖尿病组和正常对照组大鼠结肠组织中均有表达,主要分布在大鼠结肠组织的肌间丛、黏膜下丛。②胃肠推进率:糖尿病组低于正常对照组(0.7849±0.0722,0.8619±0.0481,P<0.001)。③结肠P物质阳性细胞面积:糖尿病组近端和远端结肠肌间丛均小于正常对照组犤(3.25±0.36)×104,(3.50±0.42)×104μm2;(3.26±0.34)×104,(3.47±0.26)×104μm2,P<0.01,0.05犦,而黏膜下丛分布无显著差异(P>0.05)。④P物质mRNA表达(SP/GAPDH):糖尿病组近端、远端结肠组织中表达均少于正常对照组(0.396±0.042,0.428±0.045;0.375±0.035;0.402±0.039,P<0.05)。结论:糖尿病大鼠结肠肌间丛P物质含量的减少与胃肠运动减慢存在一定相关性,且其结肠存在P物质基因表达的下调。
AIM： To observe the distribution and changes of content of substance P in colon tissue of diabetic rat with gastrointestinal dysfunction and analyze the possible mechanism in the attack of gastrointestinal dysfunction associated with diabetes mellitus.
METHODS： This experiment was carried out in the animal center of Nanjing Medical University and in the experimental room of the department of digestion of the First Affiliated Hospital of Nanjing Medical University from March to August 2005. Thirty SD rats were selected and randomly divided i0to the normal control group （n=10） and the diabetic model group （n=20）. Rats in the diabetic model group were made into diabetic models with intraperitoneal injection of streptozotocin （STZ） （80 mg/kg） for only one time, and those in normal control group were intraperitoneally injected with the equal citric acid buffer. After 72 hours, the level of blood glucose was detected in venous blood of tails. The rats with fasting blood glucose （FBG） equal to or more than 16.7 mmol/L served as diabetic rats. After raising rats of two groups for 6 weeks, the rats in the two groups were killed. The driving rate of colon was calculated under no tension. The expression of substance P in the colon tissue of rats was detected by immunohistochemistry, and the expression of substance P mRNA in the colon tissue of rats was detected by reverse transcription polymerase chain reaction （RTPCR）.
RESULTS： Twenty rats were involved in the result analysis, and 10 in each group. ①The expression of substance P-was found in the colon tissue of rats in the normal control group and diabetic model group, and the immunoreactive positive products of substance P were mainly distributed in the myenteric plexus and submucosal plexus. ②Driving rate of colon： It was lower in the diabetic model group than that in the normal control group （0.784 9±0.072 2,0.861 9±0.048 1 ,P 〈 0.001 ）. ③Area of positive cells of substance P in colon tissue： It was smaller in the myenteric plexus of proximal and distal colon in the diabetic model group than that in the normal control group [（3.25±0.36）×10^4, （3.50±0.42）×10^4 μm^2; （3.26±0.34）×10^4, （3.47±0.26）×10^4μm^2,P 〈 0.01,0.05], while there was no significant difference in the other regions （P 〉 0.05）. ④The expression of substance P mRNA（SP/GAPDH）： It in the proximal and distal colon tissue in the diabetic group was less as compared with that in the normal control group （0.396±0.042,0.428±0.045 ;0.375±0.035 ;0.402±0.039,P 〈 0.05）.
CONCLUSION： The content of substance P in the myenteric plexus of colon in the diabetic rats decreases and has relation with the decrease of movement of stomach and enteron. Besides, the gene expression of substance P in colon reduces.
出处
《中国临床康复》
CSCD
北大核心
2006年第4期94-96,i0002,共4页
Chinese Journal of Clinical Rehabilitation
基金
江苏省"一三五工程"医学重点人才基金资助项目
江苏省自然科学基金资助项目(BK2004158)~~
