miR-144在大鼠反流性食管炎中的作用和机制

The role of microRNA-144 in reflux esophagitis in rats

[目的]探讨miR-144在反流性食管炎病变中的作用和机制。[方法]取45只SD雄性大鼠随机分为假手术对照组(简称对照组)、胃食管反流病组(简称GERD组)、胃食管反流病miR-144抑制剂治疗组(简称治疗组),每组15只。GERD组、治疗组大鼠胃结扎联合幽门限制法建立胃食管反流病模型;术后第1、2、3天,对照组、GERD组分别尾静脉注射0.9%氯化钠溶液1ml,治疗组分别尾静脉注射miR-144antagomir 5OD。造模第14天处死各组大鼠,肉眼观察食管黏膜损伤情况,苏木素-伊红(HE)染色观察食管黏膜的病理改变,免疫组化法检测食管黏膜核因子E2相关因子(Nrf2)、基质金属蛋白酶3(MMP3)、基质金属蛋白酶9(MMP9)、CD68、诱导型一氧化氮合酶(iNOS)表达,并对其表达进行软件定量分析,计算平均吸光度。[结果]GERD组、治疗组大鼠食管黏膜可见反流性食管炎改变,治疗组重度反流性食管炎比率为15.38%(2/13),较GERD组46.15%(6/13)明显降低(P<0.05)。治疗组食管黏膜Nrf2表达明显高于GERD组(P<0.05),MMP3、MMP9、CD68、iNOS的表达均低于GERD组(P<0.05)。[结论]miR-144抑制剂通过增加食管黏膜Nrf2的表达,减轻炎症反应及氧化应激程度,保护食管黏膜屏障功能,从而减轻反流导致的食管黏膜损伤。

[Objective]To investigate the role and its mechanism of microRNA-144 in reflux esophagitis(RE)in rats.[Methods]A total of 45 male Sprague Dawley rats were divided into sham operation group(n=15),gastroesophageal reflux disease(GERD)group(n=15)and miR-144 antagomir-treat group(n=15).GERD rat models were established by ligating fore-stomach and constricting proximal duodenum.In one,two and three days after models were established,5 OD of miR-144 antagomir was injected into the veins of the tails in miR-144 antagomir-treat group.Rats were sacrificed 14 days later,and the esophageal tissues were histologically examined and Nrf2,MMP3,MMP9,CD68 and iNOS were immunohistochemically detected.[Results]RE present in GERD group and miR-144 antagomir-treat group.The rate of sever esophagitis in miR-144 antagomir-treat group was lower than that in GERD group(15.38% vs 46.15%,P<0.05).Compared with the GERD group,the expression of Nrf2 in esophageal mucosa was significantly increased(P<0.05),and the expressions of MMP3,MMP9,CD68 and iNOS in esophageal mucosa were significantly decreased(P<0.05)in miR-144 antagomir-treat group.[Conclusion]MiR-144 inhibitor may have anti-inflammation and anti-oxidative stress effect on RE,which is related with Nrf2 activation.