基于IL-6/miR-155轴探讨溃结宁膏穴位敷贴对脾肾阳虚型溃疡性结肠炎大鼠的影响

Effects of Acupoint Application of Kuijiening Plaster on Rats with Ulcerative Colitis Based on IL-6/miR-155 Axis

目的观察溃结宁膏穴位敷贴对脾肾阳虚型溃疡性结肠炎(UC)大鼠IL-6/miR-155轴的影响,探讨其治疗UC的作用机制。方法采用腺嘌呤、番泻叶分阶段灌胃,同时进行环境干预,结合三硝基苯磺酸灌肠法制备脾肾阳虚型UC大鼠模型。将大鼠分为空白组、模型组、非穴位敷贴组、穴位敷贴组和柳氮磺嘧啶(SASP)组,各组分别予相应处理2周。观察大鼠一般状况,对大鼠进行疾病活动指数(DAI)、结肠黏膜损伤指数(CMDI)评分,ELISA检测血清白细胞介素(IL)-8、IL-17、IL-23含量,RT-PCR检测结肠组织IL-6、IL-17、IL-23、miR-155、信号传导与转录激活因子(STAT)3 mRNA表达,Western blot检测结肠组织IL-6、p-STAT3蛋白表达,流式细胞术检测血液Th17与Treg细胞比例。结果与空白组比较,模型组大鼠DAI、CMDI评分明显升高(P<0.05),血清IL-8、IL-17、IL-23含量明显增加(P<0.05),结肠组织IL-6、IL-17、IL-23、miR-155、STAT3 mRNA表达明显升高(P<0.05),IL-6、p-STAT3蛋白表达明显升高(P<0.05),血液Th17细胞比例升高,Treg细胞比例降低(P<0.05);与模型组及非穴位敷贴组比较,穴位敷贴组和SASP组大鼠DAI、CMDI评分明显降低(P<0.05),血清IL-8、IL-17、IL-23含量明显减少(P<0.05),结肠组织IL-6、IL-17、IL-23、miR-155、STAT3 mRNA表达明显降低(P<0.05),IL-6、p-STAT3蛋白表达明显降低(P<0.05),血液Th17细胞比例降低,Treg细胞比例升高(P<0.05)。结论溃结宁膏穴位敷贴可能通过调控IL-6/miR-155轴调节UC炎症反应与Th17/Treg细胞平衡,从而治疗UC。

Objective To investigate the effects of acupoint application of Kuijiening Plaster on IL-6/miR-155 axis of ulcerative colitis(UC)rats with spleen-kidney yang deficiency;To discuss its mechanism for the treatment of UC.Methods The UC rat model of spleen-kidney yang deficiency was established by intragastric administration of adenine and senna leaf in stages,environmental intervention and TNBS enema.The rats were divided into blank group,model group,non-acupoint application group,acupoint application group and sulfasalazine(SASP)group.Each group was treated for 2 weeks.The general conditions of the rats were observed,the DAI and CMDI scores were performed on the rats,the contents of IL-8,IL-17 and IL-23 in serum were detected by ELISA,the mRNA expressions of IL-6,IL-17,IL-23,miR-155 and STAT3 in colon tissue were detected by RT-PCR,the protein expressions of IL-6 and p-STAT3 in colon tissue were detected by Western blot,and the proportion of Th17 and Treg cells in blood was detected by flow cytometry.Results Compared with the blank group,the DAI and CMDI scores significantly increased in model group(P<0.05),the contents of serum IL-8,IL-17 and IL-23 significantly increased(P<0.05),the mRNA expressions of IL-6,IL-17,IL-23,STAT3 and miR-155 significantly increased(P<0.05),the expressions of IL-6 and p-STAT3 protein increased significantly(P<0.05),the proportion of Th17 cells in blood increased,and the proportion of Treg cells decreased(P<0.05).Compared with the model group and non-acupoint application group,the DAI and CMDI scores in acupoint application group and SASP group significantly decreased(P<0.05),the contents of IL-8,IL-17 and IL-23 in serum decreased(P<0.05),the mRNA expressions of IL-6,IL-17,IL-23,STAT3 and miR-155 significantly decreased(P<0.05),the protein expressions of IL-6 and p-STAT3 significantly decreased(P<0.05),the proportion of Th17 cells in blood decreased,and the proportion of Treg cells increased(P<0.05).Conclusion Acupoint application of Kuijiening Plaster may regulate UC inflammatory response and Th17/Treg cells balance by mediating IL-6/miR-155 axis,thereby treating UC.