香叶木素对脓毒症所致小肠损伤大鼠模型的治疗作用

Therapeutic Effect of Diosmetin on Sepsis-Induced Intestinal Injury in Rats

目的探讨香叶木素(diosmetin,Dio)对脓毒症所致小肠损伤大鼠模型的治疗作用及相关机制。方法利用盲肠结扎穿刺法构建脓毒症小肠损伤大鼠模型,将大鼠随机分为对照组(术后每日后腹腔注射生理盐水20 mg·kg^(-1),连续7 d)、模型组(术后每日后腹腔注射生理盐水20 mg·kg^(-1),连续7 d)、治疗组(术后每日腹腔注射Dio 20 mg·kg^(-1),连续7 d),每组10只。HE染色法观察各组大鼠小肠组织形态学改变;激光多普勒灌注成像仪检测3组大鼠小肠黏膜下血流灌注情况;ELISA方法检测3组大鼠小肠组织中IL-1β、IL-6以及TNF-α水平;Western blot检测3组大鼠小肠组织中ZO-1、Claudin-1及NF-κB p65表达水平;3组大鼠小肠组织切片进行免疫荧光染色。结果Dio治疗可减轻脓毒症对小肠的损伤,使小肠形态得到明显改善;与模型组相比,Dio治疗可显著改善脓毒症大鼠空肠黏膜血流灌注(P<0.05);Dio治疗组大鼠小肠组织中的炎症因子(IL-1β、IL-6及TNF-α)与模型组相比均显著下降(P<0.05);与模型组相比,Dio治疗组大鼠小肠组织中ZO-1、Claudin-1及NF-κB p65水平显著下降(P<0.01);免疫荧光染色显示Dio治疗组NF-κB p65数量明显下降。结论Dio通过抑制NF-κB对脓毒症小肠损伤具有治疗保护作用。

Objective To investigate the therapeutic effect of diosmetin(Dio)on small intestinal injury induced by sepsis in rats and its mechanism of action.Methods The rat model of sepsis-induced small intestine injury was established by cecal ligation and puncture.Rats were randomly divided into three groups,with 10 rats in each group.The control,model and treatment groups were intraperitoneally injected with normal saline,normal saline and Dio at a dose of 20 mg·kg^(-1) per day for 7 consecutive days after operation,respectively.The histological changes in small intestine were observed using HE staining.Small intestinal submucosal blood perfusion was detected by laser Doppler perfusion imaging.The levels of IL-1β,IL-6 and TNF-αin small intestine were measured by ELISA.The expression of ZO-1,Claudin-1 and NF-κB p65 in small intestine was determined by Western blot and immunofluorescence staining.Results Treatment with Dio can reduce the damage of small intestine caused by sepsis and improve the morphology of small intestine.Compared with the model group,Dio treatment decreased the blood perfusion of jejunum mucosa and down-regulated the levels of IL-1β,IL-6 and TNF-αand the expression of ZO-1,Claudin-1 and NF-κB p65 expression in small intestine(P<0.01).Immunofluorescence staining showed that the number of NF-κB p65-positive cells in the treatment group was lower than that in the model group(P<0.01).Conclusion Dio can protect against sepsis-induced intestinal injury by inhibiting NF-κB activation.