艾灸“天枢”“上巨虚”对克罗恩病大鼠肠道菌群及血清炎症因子的影响

Effects of moxibustion at‘Tianshu'(ST 25)and‘Shangjuxu'(ST 37)on intestinal flora and serum inflammatory factors in rats with Crohn disease

目的:观察艾灸“天枢”“上巨虚”对克罗恩病(CD)大鼠肠道炎症损伤的影响,从肠道菌群角度探讨艾灸对CD大鼠结肠的保护作用。方法:以2,4,6三硝基苯磺酸灌肠制备CD大鼠模型。实验设正常组、模型组、艾灸组和药物组。艾灸组予艾灸双侧“天枢”和“上巨虚”,药物组予美沙拉嗪灌胃。记录大鼠体质量、粪便隐血情况并评估疾病活动指数(DAI);HE染色观察结肠病理损伤;ELISA法检测血清肿瘤坏死因子-α(TNF-α)、组胺、缓激肽水平;16S rDNA测序技术检测并分析肠道菌群组成结构的改变;冗余分析(RDA)对肠道菌群与血清炎症因子进行相关性分析。结果:与正常组比较,模型组大鼠疾病症状明显,DAI评分显著升高(P<0.05),结肠黏膜存在明显的炎症损伤,血清TNF-α、组胺、缓激肽水平显著升高(P<0.05);与模型组比较,艾灸组与药物组大鼠疾病症状缓解,DAI评分显著下降(P<0.05),结肠黏膜炎症损伤减轻,血清TNF-α、组胺、缓激肽水平显著降低(P<0.05)。16S rDNA测序显示,与正常组比较,模型组大鼠菌群丰富度和多样性减少,菌群组成结构差异较大;与模型组比较,艾灸组、药物组菌群丰富度和多样性较高,菌群结构趋于正常组。此外,考拉杆菌属、Allobaculum属、厌氧杆菌属与血清炎症因子之间呈正相关。结论:艾灸“天枢”“上巨虚”治疗CD的机制可能是通过调控肠道菌群紊乱状态,减轻肠道炎症损伤,从而维持肠道免疫稳态。

Objective:To observe the effects of moxibustion at‘Tianshu'(ST 25)and‘Shangjuxu'(ST 37)on the inflammatory injury in the colon of rats with Crohn disease(CD),so as to explore the protective mechanism of moxibustion on CD rats based on intestinal flora.Methods:The CD model was established by enema of 2,4,6 trinitrobenzene sulfonic acid(TNBS).Rats were randomly divided into normal group(NC),model group(CD),model+moxibustion group(CD+MOX),and model+mesalazine group(CD+MES).The CD+MOX group was given moxibustion at the bilateral‘Tianshu'(ST 25)and‘Shangjuxu'(ST 37),and the CD+MES group was given mesalazine gavage.Body weight and fecal occult blood of rats were recorded and disease activity index(DAI)was assessed.HE staining was performed to observe pathological damage to the colon.Tumor necrosis factor-α(TNF-α),histamine and bradykinin levels were measured by ELISA.Alterations in the composition and structure of the intestinal flora were detected and analyzed by 16S rDNA sequencing technology.Redundancy analysis(RDA)was performed to correlate intestinal flora with serum inflammatory factors.Results:Compared with the NC group,the rats in the CD group had significant disease symptoms,elevated DAI scores(P<0.05),significant infammatory damage to the colonic mucosa,and the serum levels of TNF-α,histamine and bradykinin were increased significantly(P<0.05).Compared with the CD group,the rats in the CD+MOX and CD+MES groups showed remission of disease symptoms,significant decrease in DAI score(P<0.05),reduction in colonic mucosa infiammatory injury,and significant decrease in serum TNF-α,histamine,and bradykinin levels(P<0.05).16S rDNA sequencing showed that the intestinal flora richness and diversity were reduced in the CD group compared with the NC group,and the structure of flora composition was more different;compared with the CD group,the CD+MOX and CD+MES groups had higher flora richness and diversity,and the structure of flora tended to the NC group.Additionally,there was a positive correlation between Phascolarctobacterium,Allobaculum and Anaerostipes and serum inflammatory factors.Conclusion:The mechanism of moxibustion at Tianshu'(ST 25)and‘Shangjuxu'(ST 37)in the treatment of CD may be to The mechanism of moxibustion‘Tianshu'(ST 25)and‘Shangjuxu'(ST 37)in treating CD may be to maintain intestinal immune homeostasis by regulating the state of intestinal flora disorder and reducing intestinal inflammatory damage.