瞬时受体电位通道 A1促进盆腔神经损伤小鼠结肠动力恢复的实验研究

Transient receptor potential ankyrin 1 promotes recovery of colonic motility following pelvic nerve injury in mice

目的探讨瞬时受体电位通道A1(transient receptor potential ankyrin 1,TRPAl)在去盆腔神经支配(pelvic nerve denervation,PND)小鼠结肠动力适应性恢复中的作用。方法将108只C57小鼠采用随机数字表法分为两组(n=54):PND模型组、假手术组。两组依据干预措施分别再分为3组(n=18):无干预组、TRPA1激动剂(姜黄素)处理组、TRPA1拮抗剂(HC-030031)处理组。另将30只TRPA1基因敲除小鼠随机分为2组(n=15):PND模型组、假手术组。建立PND小鼠模型,术中盲肠预置管以进行结肠传输试验。结肠传输功能分别在术后第1、3、7天测定。结果与假手术组相比,PND模型组小鼠术后第1天传输功能显著降低(P<0.001),术后第3天的结肠传输功能有恢复趋势但仍然降低(P=0.040),至第7天,两组传输功能的差异无统计学意义(P=0.073)。经TRPA1激动剂(姜黄素)干预后,与对照组(无干预假手术组)相比,干预后假手术组结肠传输功能明显增强(P<0.001),PND模型组小鼠术后第1天和第3天的结肠传输功能无差异(P=0.304、0.065),至第7天,PND模型组传输功能显著增高(P=0.001)。相反,TRPA1拮抗剂(HC-030031)干预后,PND模型组与对照组相比,小鼠术后第1、3、7天的结肠传输功能均显著下降(P<0.05),结肠传输功能无恢复趋势。对TRPA1基因敲除小鼠观察到的结果与应用TRPA1拮抗剂的结果一致。免疫组化结果显示:TRPA1表达于小鼠结肠黏膜,小鼠去盆腔神经支配后,TRPA1出现先下降、后逐渐升高的适应性恢复趋势。结论小鼠去盆腔神经支配后存在结肠动力适应性恢复现象,TRPA1表达于结肠黏膜,可显著促进PND小鼠的结肠动力恢复。

Objective To investigate the role of transient receptor potential ankyrin 1(TRPA1) in the recovery of colonic motility following pelvic nerve denervation(PND) in mice. Methods A total of 108 male C57 mice were randomized into PND model group(n=54) and sham operation group(n=54), and in each group, the mice were further randomized into untreated group, TRPA1 agonist(curcumin) treatment group, and TRPA1 antagonist(HC-030031) treatment group. Another 30 male TRPA1 knockout mice were also randomized into PND group(n=15) and sham operation group(n=15). All the mice underwent open surgery for PND by severing the pelvic nerve or received sham operation, and during the operation, a silicone tube was embedded in the cecum near the colon for testing colonic transit function on postoperative days 1, 3, and 7. Results Compared with the sham-operated mice, the PND mice showed significantly decreased colonic transit function on postoperative days 1(P<0.001) and 3(P=0.040) but had comparable transit function on day 7. Compared with the sham-operated mice without any interventions(control group), curcumin treatment resulted in significantly enhanced colonic transit function in the sham-operated mice(P<0.001), and promoted nearly full recovery of colonic transit function in PND mice on postoperative days 1(P=0.304 vs control) and 3(P=0.304 vs control) and even further enhancement on day 7(P=0.001). In PND mice, treatment with the TRPA1 antagonist HC-030031 significantly lowered colonic transit function(P<0.05) without any signs of recovery. The results of colonic transit test in TRPA1 knockout mice were consistent with those in HC-030031-treated PND mice. The results of immunohistochemistry showed that TRPA1 was expressed in the colonic mucosa of the mice, and PND caused a transient decrease in TRPA1 expression followed by a gradual recovery. Conclusion The colonic motility shows an adaptive recovery mechanism after PND in mice. TRPA1, which is expressed in the colonic mucosa of mice, can significantly promote colonic motility recovery following PND in mice.