目的：观察隔药灸对克罗恩(Crohn’s Disease,CD)大鼠结肠p38MAPK、ERK1/2、c-fos的影响,探讨隔药灸治疗CD的作用机制。方法：将清洁级雄性SD大鼠随机分为正常组、模型组、隔药灸组和假灸组4组。采用TNBS合50%乙醇溶液灌肠制备大鼠CD模型。模型制备成功后,隔药灸组取天枢穴(双)、气海穴进行隔药饼灸治疗；假灸组取穴、操作与隔药灸组相同,但不点燃艾炷。治疗结束后,采用HE染色,光镜下观察大鼠结肠组织形态学变化；采用实时荧光定量PCR技术检测结肠p38MAPK mRNA表达；应用ELISA技术检测结肠p38MAPK、c-fos蛋白含量,Western blot技术检测结肠ERK1/2蛋白表达。结果：与正常组比较,模型组大鼠结肠组织损伤严重,可见全壁性炎症、裂隙状溃疡,部分大鼠结肠可见纤维化；与模型组、假灸组比较,隔药灸组大鼠结肠形态结构改善,肠道炎症减轻；假灸组大鼠结肠损伤程度、炎症反应与模型组类似。与正常组比较,模型组p38MAPK mRNA表达增加(P＜0.01),p38MAPK、ERK1/2、c-fos蛋白含量均增加(均Ｐ＜0.05)；与模型组比较,隔药灸组p38MAPK mRNA表达降低(P＜0.05),p38MAPK、ERK1/2、c-fos蛋白含量均降低(均Ｐ＜0.05)；假灸组均无显著变化(均Ｐ＞0.05)。结论：隔药灸能下调克罗恩病大鼠结肠p38MAPK、ERK1/2、c-fos的表达,改善炎症、促进肠道组织修复。
Objective To explore the action mechanism of herb-partitioned moxibustion (HPM) in treating Crohn's disease (CD) by observing the effect of HPM on colonic p38MAPK, ERK1/2 and c-fos. Method Male Sprague Dawley (SD) rats of clean grade were randomly divided into 4 groups: normal group (NG), model group (MG), HPM group (HPMG) and sham moxibustion group (SMG). The CD rat models were developed using the mixture of 2,4,6-trinitrobenzene sulfonic acid (TNBS) and alcohol via enema. When the models were successfully established, the HPMG received HPM at bilateral Tianshu (ST25) and Qihai (CV6) points; the selection of acupoints and operation in the SMG were the same, except that the moxa cones were not ignited. After treatment, the histopathological changes in rat colons were observed by HE staining under light microscope; the p38MAPK mRNA expression in rat colons was detected by real-time quantitative PCR; the protein contents of colonic p38MAPK and c-fos were detected by using ELISA; the protein expression of colonic ERK1/2 was measured by Western blot. Result Compared with the NG, the colon tissues were severely damaged in the MG, presenting inflammation in the whole wall and fissure-like ulcers, as well as fibrosis in some rats; compared with the MG and SMG, the colonic morphology was improved and the intestinal inflammation was alleviated in the HPMG; the degree of colonic injury and inflammatory response in the SMG were similar to those in the MG. Compared with the NG, the expression of p38MAPK mRNA increased significantly (P＜0.01), and the protein expressions of p38MAPK, ERK1/2 and c-fos also increased significantly (all P＜0.05) in the MG. Compared with the MG, the expression of p38MAPK mRNA decreased significantly (P＜0.05), and the protein expressions of p38MAPK, ERK1/2 and c-fos also dropped significantly (P＜0.05) in the HPMG. Compared with the MG, there were no significant changes in the indexes in the SMG (P＞0.05). Conclusion HPM can down-regulate the expressions of colonic p38MAPK, ERK1/2 and c-fos, so as to improve the intestinal inflammation and promote tissue repair in CD rats.