隔药饼灸对克罗恩病肠纤维化大鼠结肠氧化应激和Nrf2的调节作用  

Regulatory effects of herb‑partitioned moxibustion on colonic oxidative stress and Nrf2 expression in rats with Crohn's disease‑induced intestinal fibrosis

作　　者：张智英 赵敏 李琪[1] 韩冬 吴焕淦[1,2] 乔宇 郑寒丹 翁志军[2] 张方 刘慧荣[1,2] ZHANG Zhiying;ZHAO Min;LI Qi;HAN Dong;WU Huangan;QIAO Yu;ZHENG Handan;WENG Zhijun;ZHANG Fang;LIU Huirong(Key Laboratory of Acupuncture Immune Effects,Yueyang Hospital of Integrated Traditional Chinese and Western Medicine Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 200437,China;Shanghai Research Institute of Acupuncture and Meridian,Shanghai 200030,China)

机构地区：[1]上海中医药大学附属岳阳中西医结合医院针灸免疫效应重点研究室,上海200437 [2]上海市针灸经络研究所,上海200030

出　　处：《上海中医药杂志》2024年第12期163-169,共7页Shanghai Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(81873374);国家中医药管理局高水平中医药重点学科建设项目(zyyzdxk-2023068);上海市自然科学基金项目(21ZR1460000);上海市科委科技计划项目(20MC1920500)。

摘　　要：目的观察隔药饼灸对克罗恩病(CD)肠纤维化大鼠结肠氧化应激标志物和核因子E2相关因子2(Nrf2)表达的影响,以探究其治疗CD肠纤维化的效应机制。方法将SD大鼠分为正常组、模型组、隔药饼灸组,每组各6只,通过三硝基苯磺酸(TNBS)与乙醇混合液灌肠制备CD肠纤维化大鼠模型(每周1次,连续6周)。隔药饼灸组选取大鼠“天枢穴”和“气海穴”进行隔药饼灸治疗(每日1次,连续10 d)。治疗结束后,测量大鼠结肠长度,观察结肠病理形态表现;Western blot法检测结肠α-平滑肌肌动蛋白(α-SMA)、胶原蛋白Ⅰ(ColleganⅠ)表达;硫代巴比妥酸(TBA)法检测结肠丙二醛(MDA)含量、羟胺法检测结肠超氧化物歧化酶(SOD)活性、比色法检测谷胱甘肽过氧化物酶(GSH-Px)活性;酶联免疫吸附测定(ELISA)及实时荧光定量逆转录聚合酶链式反应(RT-qPCR)法检测结肠Nrf2蛋白及其mRNA表达。结果①模型组大鼠结肠长度较正常组显著短缩(P<0.05);隔药饼灸组大鼠结肠长度较模型组明显增加(P<0.05)。②模型组大鼠结肠组织较正常组结构紊乱,并可见CD典型裂隙样溃疡及大量胶原纤维组织沉积(P<0.05);隔药饼灸组大鼠结肠组织较模型组排列整齐,溃疡组织表层可见新生上皮细胞修复,结肠胶原纤维组织沉积明显减少(P<0.05)。③模型组大鼠结肠α-SMA、Collegan I、MDA表达水平较正常组明显增高(P<0.05);隔药饼灸组大鼠结肠α-SMA、Collegan I、MDA蛋白表达较模型组明显降低(P<0.05)。④模型组大鼠结肠SOD、GSH-Px、Nrf2表达较正常组明显降低(P<0.05);隔药饼灸组大鼠结肠SOD、GSH-Px、Nrf2表达较模型组均明显增高(P<0.05)。结论隔药饼灸能够抑制CD大鼠肠纤维化,缓解肠道炎症及氧化损伤,促进结肠组织修复,其机制可能与隔药饼灸能够调节Nrf2并转导其下游相关抗氧化物酶有关。Objective To investigate the effects of herb-partitioned moxibustion on oxidative stress markers and Nrf2 expression in the colons of rats with Crohn's disease(CD)-induced intestinal fibrosis,and to explore its therapeutic mechanism for treating CD-induced intestinal fibrosis.Methods SD rats were randomly divided into three groups(n=6 per group):normal group,model group,and herb-partitioned moxibustion group.CD-induced intestinal fibrosis was modeled in rats using TNBS-ethanol enema(once a week for 6 weeks).The herb-partitioned moxibustion group received moxibustion at"Tianshu(ST25)"and"Qihai(CV6)"acupoints(once daily for 10 consecutive days).After treatment,colon length was measured,and histopathological changes were observed.Western blot was used to detect the expression of alphasmooth muscle actin(α-SMA)and Collagen I(Col I)in the colon.The thiobarbituric acid(TBA)method was used to detect the content of malondialdehyde(MDA),the hydroxylamine method was used to detect superoxide dismutase(SOD)activity,and the colorimetric method was used to detect glutathione peroxidase(GSH-Px)activity.Nrf2 protein and mRNA expression in the colon were detected using Nrf2-linked enzyme-linked immunosorbent assay(ELISA)and real-time quantitative reverse transcription polymerase chain reaction(RT-qPCR).Results①The colon length in the model group was significantly shorter than that in the normal group(P<0.05);however,the colon length in the herb-partitioned moxibustion group was significantly longer than that in the model group(P<0.05).②Compared with the normal group,the model group showed disorganized colon tissue structure,with fissure-like ulcers and significant collagen fiber deposition(P<0.05).Compared with the model group,the herb-partitioned moxibustion group showed more intact colon tissue,with new epithelial cell repair on the ulcer surface and significantly reduced collagen fiber deposition(P<0.05).③Compared with the normal group,the model group had significantly higher expression levels ofα-SMA,Col I,and MDA in

关 键 词：克罗恩病 肠纤维化 隔药饼灸 艾灸 氧化应激 核因子E2相关因子2 作用机制 

分 类 号：R245[医药卫生—针灸推拿学]