复方芩柏颗粒剂通过靶向干预miR-199-3p对溃疡性结肠炎的改善作用

Improvement of ulcerative colitis by targeted intervention of miR-199-3p with Compound Qinbai Granule

目的观察复方芩柏颗粒剂通过靶向干预miR-199-3p抑制磷脂酰肌醇3-激酶(PK3K)/蛋白激酶B(AKT)-雷帕霉素靶蛋白(m TOR)信号通路激活对溃疡性结肠炎(UC)的作用。方法用5%2,4,6-三硝基苯磺酸(TNBS)与无水乙醇按1∶1体积制成TNBS/乙醇混合液,建立溃疡性结肠炎模型。用随机数字表法将大鼠随机分为3组:正常组、模型组和实验组,每组19只。正常组和模型组给予0. 9%Na Cl 2 m L灌肠,实验组给16%复方芩柏颗粒2 m L灌肠,2次/天,连续3周。以免疫组化法检测结肠组织中m TOR表达,以蛋白质印迹法检测结肠组织PI3K/AKT相关蛋白表达,以实时荧光定量-PCR法测定结肠组织miR-199-3p基因表达。结果正常组、模型组和实验组的m TOR阳性表达(平均光密度)分别为0. 18±0. 06,0. 67±0. 09和0. 40±0. 06;上述3组的PI3KP85蛋白相对表达量分别为0. 18±0. 04,0. 83±0. 09和0. 58±0. 08;上述3组的PI3KP100蛋白相对表达量分别为0. 19±0. 05,0. 71±0. 11和0. 54±0. 08;上述3组的p-AKT蛋白相对表达量分别为0. 21±0. 06,0. 79±0. 05和0. 49±0. 08;上述3组大鼠结肠组织miR-199-3p基因相对表达量分别为0. 62±0. 09,0. 19±0. 0和0. 38±0. 09。上述指标:模型组和正常组相比较差异均有统计学意义(均P <0. 05);实验组与模型组比较差异均有统计学意义(均P <0. 05)。结论增加miR-199-3p表达和抑制PI3K/AKT-m TOR信号通路激活可能是复方芩柏颗粒剂治疗UC的重要机制。

Objective To observe the effect of Compound Qinbai Granule on ulcerative colitis(UC)by targeting microRNA-199-3 p(miR-199-3 p)on phosphatidylinositol 3-kinase(PK3 K)/protein kinase B(AKT)-mammalian target of rapamycin(mT OR)signaling pathway activation.Methods The 2,4,6-trinitrobenzene sulfonic acid(TNBS)/ethanol mixture was prepared by 1∶1 volume with 5%TNBS/ethanol mixture for establishment of ulcerative colitis model.SD rats were divided into three groups:normal group,model group and experimental group,19 rats in each group.Model group and normal group were given the same amount of saline,and experimental group was given 16%Compound Qinbai Granule 2 mL enema,twice a day for 3 weeks.Expression of mTOR in colon tissue was detected by immunohistochemistry,expression of PI3 K/AKT-related protein in colon tissue was detected by Western blot,and expression of miR-199-3 p in colon tissue was detected by real time fluorescence quantitative-PCR.Results Positive expression(average optical density)of mT OR in normal group,model group and experimental group were0.18±0.06,0.67±0.09,0.40±0.06;relative expression of PI3 KP85 protein in the above three groups were0.18±0.04,0.83±0.09,0.58±0.08;relative expression of PI3 KP100 protein in the above three groups were0.19±0.05,0.71±0.11,0.54±0.08;relative expression levels of p-AKT protein in the three groups were0.21±0.06,0.79±0.05,0.49±0.08;relative expression levels of miR-199-3 p in colon tissues of the three groups were 0.62±0.09,0.19±0.08,0.38±0.09.Comparison between model group and normal group,the difference of the factors were significantly(all P<0.05);comparison between experimental group and model group,the difference of the factors were significantly(all P<0.05).Conlusion Increasing the expression of miR-199-3 p and inhibiting the activation of PI3 K/AKT-mTOR signaling pathway may be an important mechanism of Compound Qinbai Granule in treating UC.