微RNA-490-5p对肠功能紊乱大鼠内脏敏感性的影响  

Effects of microRNA-490-5p on visceral sensitivity in rat with intestinal dysfunction

作　　者：罗和生[1] 任海霞[1] 张法灿[2] 梁列新[2] 张国[2] 

机构地区：[1]武汉大学人民医院消化内科,430060 [2]广西壮族自治区人民医院消化内科

出　　处：《中华消化杂志》2017年第4期254-259,共6页Chinese Journal of Digestion

基　　金：国家自然科学基金（81160053）

摘　　要：目的探讨微RNA-490-5p在调控肠功能紊乱大鼠内脏高敏感中的作用。方法用慢病毒载体系统构建微RNA-490-5p的静默载体，将大鼠分为正常组、腹泻型IBS组、慢病毒空载体组和静默组，其中腹泻型IBS组、慢病毒空载体组和静默组为模型组。实时荧光定量PCR法检测静默效率，通过10％印度墨汁灌胃计算首次黑便排出时间、粪便含水率，引起腹部抬高（或背部拱起）的扩张容量阈值，以不同强度的结肠扩张刺激下大鼠的腹部撤回反射（AwR）评分来判断静默微RNA-490-5p后大鼠内脏的敏感性，以BL-420F生物机能实验系统检测大鼠结肠扩张刺激下腹壁肌电活动和氯化乙酰胆碱刺激下大鼠离体结肠肠管张力的改变。采用t检验比较模型组和正常组间的差异，慢病毒干预后多组间比较采用单因素方差分析，当方差齐时多组间两两比较用最小显著差法（LSD），方差不齐时用Games-Howell法。结果模型组胃肠推进时间和引起腹部抬高（或背部拱起）的扩张容量阈值均低于正常组[（8．54±4．07）h比（12．33±2．23）h，（0．56±0．08）mL比（O．84±0．09）mL]，差异均有统计学意义（t=2．62、6．37，P均〈O．05）。模型组0．8mL和1．2mL0．9％氯化钠溶液扩张刺激时的AWR评分均高于正常组[（3．20±0．56）分比（1．20±0．45）分，（3．73±0．46）分比（2．60±0．55）分]，差异均有统计学意义（t=7．20、4．58，P均〈0．01）；1．6mL0．9％氯化钠溶液扩张刺激时，模型组和正常组AWR评分[（3．93±0．26）分比（3．80±0．45）分]比较差异无统计学意义（P〉0．05）。静默微RNA-490-5p后，正常组、腹泻型IBS组、慢病毒空载体组和静默组大鼠胃肠推进时间分别为（11．12±1．01）、（6．23±3．17）、（6．09±2．26）和（12．36±1．97）h，4组比较差异有统计学意义（F=10．55，P〈0．01）。给予0．8mL和1．2mL0Objective To investigate the role of microRNA-490-5p （miR-490-5p） in the regulation of visceral sensitivity in rats with intestinal dysfunction. Methods The lentivirus vector system was used to construct the rno-miRNA-490-5p lentivirus expression vector. The rats were divided into normal group,diarrhea-predominant irritable bowel syndrome （IBS-D） group, lentivirus empty vector group and the siRNA silent group and the latter three groups were model groups. The efficiency of siRNA was measured by real-time polymerase chain reaction （PCR）. The rats were gavaged with 10% India ink, and then the time of first black stool, water content of feces and threshold of expansion capacity caused abdominal elevation or back arching were calculated. The visceral sensitivity of rats after miRNA-490-5p silenced was evaluated with abdominal withdrawal reflex （AWR） score by stimulating with different intensities of colonic dilatation. The abdominal electrical activity of rats stimulated by colonic distension was measured by BL-420F biological and functional experimental system. The change of the tension of rats isolated colon intestinal stimulated with aeetylcholine chloride was also detected by BL-420F biological and functional experimental system. T test was used to compare the differences between the model groups and the normal group. One way analysis Of variance was performed for muhi-group comparison after miRNA-490-5p interfered. For comparison between two groups among multiple groups, least significant difference （LSD） method was used when the variance was equal, and Games-Howell method was used when the variance was unequal. Results The gastrointestinal propulsion time and the threshold of expansion capacity caused abdominal elevation or hack arching of model groups were both lower than those of the normal group （（8.54±4.07） hvs （12.33±2.23） h, （0.56±0.08） mL vs （0.84±0.09） mL）, and the differences were statistically significant （t= 2. 62 and 6. 37, both P〈0. 05）. After diste

关 键 词：肠易激综合征 内脏敏感性 腹壁肌电 结肠张力 大鼠 

分 类 号：R574.4[医药卫生—消化系统]