不同灸量对溃疡性结肠炎大鼠结肠上皮形态学及血清中炎性细胞因子、结肠组织中炎性细胞信号转导通路的影响

目的:从形态学、免疫学、分子生物学角度探讨不同灸量治疗溃疡性结肠炎(UC)效果的异同。方法:32只SD大鼠随机分为空白组6只、模型复制组26只。采用三硝基苯甲酸/葡聚糖硫酸钠制备UC大鼠模型。模型复制成功后的大鼠按随机数字表分为模型组、3壮组、6壮组、9壮组,每组各6只。各治疗组所取穴位为"天枢""大横",艾炷直接灸法,每次分别施灸3壮(3min)、6壮(6min)、9壮(9min),共治疗14次。观察大鼠治疗前后疾病活动指数(DAI),电镜、光镜观察结肠组织形态学改变,酶联免疫法检测大鼠血清中白介素-8(IL-8)、白介素-10(IL-10)含量,Western blot法检测大鼠结肠中Toll样受体9(TLR-9)和核转录因子-κB(NF-κB)p 65表达。结果:灸法可明显降低大鼠DAI的评分(与模型组比较均P0.05);光镜与电镜结果显示,灸量越大,结肠组织腺体排列越规则。模型组血清IL-8含量升高,IL-10含量降低;与模型组比较,各治疗组IL-8降低,IL-10增高,其中9壮组和6壮组的变化较3壮组更明显(均P0.05)。模型组结肠组织中TLR-9、NF-κB p 65大量表达;与模型组比较,各治疗组TLR-9、NF-κB p 65表达均降低,且以9壮组的变化最为明显(均P0.05)。结论:艾灸可修复UC大鼠受损黏膜上皮,抑制血清中IL-8含量,提高血清中IL-10含量,通过抑制结肠组织中NF-κB p 65转录而下调TLR-9表达。灸量越大治疗效果越明显。

Influence of Moxibustion with Different Duration on Colonic Epithelial Structure,Serum Inflammatory Cytokines,and Intestinal Mucosa Inflammatory Cell Signal Transduction Pathways

Objective To observe the effect of moxibustion at different duration on colonic mucosal morphological changes,serum and colonic cytokine levels in ulcerative colitis(UC)rats,so as to provide experimental evidence for clinical treatment of UC.Methods SD rats were randomly divided into blank control,UC model,3cones-moxibustion(3-cones-M),6-cones-M and 9-cones-M groups,with 6rats in each group.UC model was established by intra-rectal injection of mixture solution of5%trinitro-benzene-sulfonic acid(TNBS,100mg/kg)and 50% alcohol(1∶1)under anesthesia and oral administration of 5%dextran sodium sulfate.Moxibustion(ignited moxa cones)was applied to"Tianshu"(ST 25)and"Daheng"(SP 15),once daily in the first 7days,and once every other day in the subsequent 14days.Serum IL-8and IL-10contents were assayed by ELISA and colonic toll-like receptor 9(TLR-9)and nuclear factor-κB p 65(NF-κB p 65)protein expression levels detected by Western blot.The colonic mucosal structure was observed by light microscope after H.E.staining,and by electron microscope,respectively.Results In comparison with the blank control group,the Disease Activity Index(DAI),serum IL-8content,colonic TLR-9and NF-κB p 65protein expression levels were significantly increased in the model group(P<0.05),and serum IL-10content was notably decreased in the model group(P<0.05).While in comparison with the model group,the DAI,serum IL-8content,colonic TLR-9and NF-κB p 65protein expression levels in the 3-cones-M,6-cones-M and 9-cones-M groups were remarkably down-regulated(P<0.05),and serum IL-10contents considerably up-regulated in the three moxibustion groups(P<0.05).No significant differences were found among the three moxibustion groups in the DAI(P>0.05).The serum IL-8contents were significantly lower and serum IL-10contents were considerably higher in the 6-cones-M and 9-cones-M groups than in the 3-cones-M group(P<0.05).The changes of colonic TLR-9and NF-κB p 65protein expression were more remarkable in the 9-cones-M group than in the 3-cones-M and 6-cones-M groups(P<0.05).Results of H.E.staining and electron microscopy showed that in the model group,mucosal injury,partial disorganization of the glandular organ,edema and congestion and inflammatory cell infiltration,mucosal epithelial microvili injury with disordered arrangement,etc.under light microscope,and local mucosal defect,apoptotic bodies and mucolysis under electron microscope were found,these situations were obviously lighter in rats of the three moxibustion groups,particularly in the 9-cones-M group.Conclusion Moxibustion intervention can relieve colonic mucosal injury in UC mice,which may be closely associated with its effects in suppressing serum proinflammatory cytokine IL-8,up-regulating anti-inflammatory cytokine IL-10level,and inhibiting colonic NF-κB p 65and TLR-9protein expression,and the effects of longer duration of moxibustion are better.