去盆腔神经对大鼠结肠动力与结肠黏膜瞬时受体电位通道A1的影响

Effects of pelvic nerve transection on the colonic motility and the expression of transit receptor potential ankyrin 1 in a rat model

目的 探讨去盆腔神经对大鼠结肠动力与结肠黏膜瞬时受体电位通道A1（TRPA1）的影响.方法 将96只成年大鼠按随机数字表法分为3组：（1）对照组18只,大鼠不做特殊处理,仅常规饲养.（2）假手术组39只,大鼠开腹后剥离出盆腔神经,缝合切口,关闭腹腔.（3）手术组39只,大鼠开腹后分离出盆腔神经,显微剪剪断,缝合切口,关闭腹腔.通过计算各组大鼠术后第1、3、7天结肠组织和粪便中铬（51Cr）分布的几何中心（GC）检测结肠传输功能.运用Western blot检测各组大鼠术后第1、3、7天近端及远端结肠黏膜TRPA1的表达.符合正态分布的计量资料以（x）±s表示,多组间比较采用重复测量的方差分析,两两比较采用LSD检验.结果 假手术组大鼠51Cr的GC值术后第1、3、7天分别为5.8±0.9、7.5±0.5、7.3±0.5,3者比较,差异有统计学意义（F =9.508,P＜0.05）.手术组大鼠51Cr的GC值术后第1、3、7天分别为4.9±0.4、5.6±0.4、6.4±0.8,3者比较,差异有统计学意义（F=11.689,P＜0.05）.假手术组和手术组大鼠51Cr的GC值术后第1、3天分别比较,差异均有统计学意义（=2.227,7.144,P＜0.05）;术后7天两组比较,差异无统计学意义（t=2.162,P＞0.05）.Western blot检测结果显示：对照组大鼠近端结肠黏膜中TRPA1相对表达量术后第1、3、7天分别为1.00 ±0.05、1.00±0.07、1.00±0.06,3者比较,差异无统计学意义（F=0.055,P＞0.05）.假手术组大鼠近端结肠黏膜中TRPA1相对表达量术后第1、3、7天分别为0.78 ±0.09、0.94 ±0.08、0.95 ±0.12,3者比较,差异有统计学意义（F=5.651,P＜0.05）.手术组大鼠近端结肠黏膜中TRPA1相对表达量术后第1、3、7天分别为0.37 ±0.12、0.89 ±0.10、0.92 ±0.14,3者比较,差异有统计学意义（F=41.005,P＜0.05）.对照组、假手术组和手术组大鼠近端结肠黏膜中TRPA1相对表达量术后第1天比较,差异有统计学意义（F=73.497,P＜0.05）;对照组分别与假手术组和手术组比较,差异均有统计学意义（t=4.224,11.954,P＜0.05）;手术组和假手术组比较,差异有统计学意义（t=7.730,P＜0.05）.对照组、假手术组和手术组大鼠近端结肠黏膜中TRPA1相对表达量术后第3、7天比较,差异均无统计学意义（F=2.087,0.656,P＞0.05）.对照组大鼠远端结肠黏膜中TRPA1相对表达量术后第1、3、7天分别为1.00±0.05、1.00±0.07、1.00±0.06,3者比较,差异无统计学意义（F=0.055,P＞0.05）.假手术组大鼠远端结肠黏膜中TRPA1相对表达量术后第1、3、7天分别为0.68 ±0.11、0.98±0.12、1.11 ±0.16,3者比较,差异有统计学意义（F=16.975,P＜0.05）.手术组大鼠远端结肠黏膜中TRPA1相对表达量术后第1、3、7天分别为0.39±0.12、0.78 ±0.10、1.06 ±0.13,3者比较,差异有统计学意义（F=50.417,P＜0.05）.对照组、假手术组和手术组大鼠远端结肠黏膜中TRPA1相对表达量术后第1、3天比较,差异均有统计学意义（F =58.773,8.680,P ＜0.05）.术后第1天,对照组分别与假手术组和手术组比较,差异均有统计学意义（t=5.706,10.837,P＜0.05）;手术组和假手术组比较,差异有统计学意义（t=5.131,P＜0.05）.术后第3天,对照组与假手术组比较,差异无统计学意义（t=0.166,P＞0.05）;对照组与手术组比较,差异有统计学意义（t=3.694,P＜0.05）;假手术组和手术组比较,差异有统计学意义（t=3.528,P＜0.05）.对照组、假手术组和手术组大鼠远端结肠黏膜中TRPA1相对表达量术后第7天比较,差异无统计学意义（F=1.319,P＞0.05）.结论 去盆腔神经支配后,大鼠结肠动力减弱,远端结肠黏膜中TRPA1表达下调,但随时间推移可逐渐恢复,两者在恢复趋势上存在一致性.

Objective To investigate the effects of pelvic nerve transection on the colonic motility and the expression of transit receptor potential ankyrin 1 （TRPA1） in the colon mucosa.Methods Ninety-six Sprague-Dawley rats were divided into 3 groups based on a random number table：（1） 18 rats in the control group remained untreated and were fed regularly;（2） 39 rats in the sham operation group received laparotomy,and the pelvic nerves were stripped;（3） 39 rats in the operation group received laparotomy with pelvic nerve transection before abdominal closure.Colonic transit was assessed respectively at postoperative day 1,3,and 7 by injecting and calculating the geometric center （GC） of the distribution of 51Cr after 3 hours of propagation.The expression of TRPA1 in the colonic mucosa was determined by Western blot at postoperative day 1,3,7.Data with normal distribution were expressed by （x）± s,and were analyzed using the repeated measures ANOVA or LSD test.Results The GC values of the distribution of 51Cr in the sham operation group at postoperative day 1,3,7 were 5.8 ± 0.9,7.5 ± 0.5,7.3 ± 0.5,with a significant difference （F =9.508,P 〈 0.05）.The GC values of the distribution of 51Cr in the operation group at postoperative day 1,3,7 were 4.9 ± 0.4,5.6 ± 0.4,6.4 ± 0.8,with a significant difference （F =11.689,P 〈 0.05）.There were significant differences in the GC values of the distribution of 51 Cr at postoperative day 1 and 3 between the sham operation group and the operation group （t =2.227,7.144,P 〈 0.05）,while no significant difference was detected at postoperative day 7 （t =2.162,P 〉 0.05）.The results of Western blot showed that the relative expressions of TRPA1 in the proximal part of the colon at postoperative day 1,3,7 were 1.00 ± 0.05,1.00 ± 0.07,1.00 ± 0.06 in the control group,with a significant difference （F =0.055,P 〉 0.05）.The relative expressions of TRPA1 in the proximal part of the colon at postoperative day 1,3,7 were 0.78 ± 0.09,0.94 ± 0.08,0.95 ± 0.12 in the sham operation group,with a significant difference （F =5.651,P 〈 0.05）.The relative expressions of TRPA1 in the proximal part of the colon at postoperative day 1,3,7 were 0.37 ± 0.12,0.89 ± 0.10,0.92 ± 0.14 in the operation group,with a significant difference （F =41.005,P 〈0.05）.There was significant difference in the relative expressions of TRPA1 in the proximal part of the colon among the 3 groups at postoperative day 1 （F =73.497,P 〈 0.05）,while significant differences were respectively detected between the control group and the sham operation group and the operation group at postoperative day 1 （t =4.224,11.954,P 〈 0.05）,and significant difference between the operation group and the sham operation group was also observed （t =7.730,P 〈 0.05）.There was no significant difference in the relative expression of TRPA1 in the proximal part of the colon among the 3 groups between day 3 and day 7 （F =2.087,0.656,P 〉 0.05）.The relative expressions of TRPA1 in the distal part of the colon at postoperative day 1,3,7 were 1.00 ± 0.05,1.00 ± 0.07,1.00 ± 0.06 in the control group,with no significant difference （F =0.055,P 〉 0.05）.The relative expressions of TRPA1 in the distal part of the colon at postoperative day 1,3,7were 0.68 ±0.11,0.98 ±0.12,1.11 ±0.16 in the sham operation group,with a significant difference （F =16.975,P 〈 0.05）.The relative expressions of TRPA1 in the distal part of the colon at postoperative day 1,3,7 were 0.39 ± 0.12,0.78 ± 0.10,1.06 ± 0.13 in the operation group,with a significant difference （F =50.417,P 〈 0.05）.There were significant differences in the relative expression of TRPA1 in the distal part of the colon among the 3 groups between day 1 and day 3 （F =58.773,8.680,P 〈 0.05）,while significant differences were respectively detected between the control group and the sham operation group and the operation group at postoperative day 1 （t =5.706,10.837,P 〈 0.05）,and significant difference was also detected between the operation group and the sham operation group （t =5.131,P 〈 0.05）.There was no significant difference in the relative expression of TRPA1 in the distal part of the colon between the control group and the sham operation group at postoperative day 3 （t =0.166,P 〉 0.05）,while significant differences were respectively detected between the control group and the operation group and between the sham operation group and the operation group at postoperative day 3 （t =3.694,3.528,P 〈 0.05）.There was no significant difference in the relative expression of the TRPA1 in the distal part of the colon between the 3 groups at postoperative day 7 （F =1.319,P 〉 0.05）.Conclusions Injury to pelvic nerves adversely affects colonic transit and expression of TRPA1 in mucosa.With a compensatory mechanism from the intestinal itself,these alterations in intestinal motility function normalize over time suggesting expression of TRPA1 in mucosa plays a crucial role in the recovery of intestinal motility function.