摘要
[目的]探究中药灌肠方对葡聚糖硫酸钠(dextran sulfate sodium,DSS)诱导的溃疡性结肠炎(ulcerative colitis,UC)的保护作用及其作用机制。[方法]给予C57BL/6J小鼠自由饮用2.5%DSS水溶液7 d以诱导UC。2 d后给药组分别给予低、中、高剂量(1.3、2.6、5.2 g/kg)的中药灌肠方和阳性药柳氮磺吡啶0.2 g/kg灌肠,模型组和正常组给予等体积0.9%氯化钠溶液灌肠。以苏木精-伊红(hematoxylin-eosin,HE)染色观察小鼠结肠组织切片病变程度;采用酶联免疫吸附试验(enzyme linked immunosorbent assay,ELISA)测定血清炎症因子肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素-6(interleukin-6,IL-6)、白细胞介素-1β(interleukin-1β,IL-1β)及白细胞介素-18(interleukin-18,IL-18)水平;采用生理生化检测试剂盒检测髓过氧化物酶(myeloperoxidase,MPO)、超氧化物歧化酶(superoxide dismutase,SOD)活性,一氧化氮(nitric oxide,NO)、丙二醛(malondialdehyde,MDA)水平;免疫印迹法检测小鼠结肠组织闭锁小带蛋白-1(zonula occludens protein-1,ZO-1)、闭合蛋白(occludin)、核苷酸结合寡聚结构域样受体蛋白3(nucleotide-binding oligomerization domain-like receptor protein 3,NLRP3)、沉默调节蛋白6(sirtuin 6,SIRT6)蛋白表达水平。[结果]与正常组比较,DSS可诱导结肠炎症、隐窝丢失、杯状细胞耗竭、炎性浸润;血清TNF-α、IL-6、IL-1β及IL-18炎症因子水平,MPO活性和NO、MDA水平明显升高(P<0.01,P<0.001),SOD活性明显降低(P<0.01)。与模型组比较,中药灌肠方治疗后明显改善DSS诱导的结肠炎症隐窝丢失、杯状细胞耗竭、炎性浸润的现象;血清TNF-α、IL-6、IL-1β及IL-18炎症因子水平,MPO活性和NO、MDA水平明显降低(P<0.01,P<0.001),中剂量和高剂量组SOD活性明显升高(P<0.05)。中药灌肠方组小鼠结肠组织ZO-1、occludin、SIRT6的蛋白表达水平明显升高(P<0.001,P<0.01,P<0.05),中剂量和高剂量组NLRP3蛋白表达水平明显降低(P<0.01)。[结�
[Objective]To investigate the protective effect of herbal enema formula on dextran sodium sulfate(DSS)-induced ulcerative colitis(UC)and its mechanism of action.[Methods]C57BL/6J mice were given 2.5%DSS aqueous solution for 7 d to induce UC.After 2 d,mice in herbal enema formula low,medium and high dose groups were given 1.3 g/kg,2.6 g/kg and 5.2 g/kg herbal enema formula for edema;0.2 g/kg sulfasalazine were given to positive control group for enema;mice in model group and normal group were given equal volumes of 0.9%sodium chloride solution for enema,respectively.The degree of lesions was observed by hematoxylin-eosin(HE)staining in mice colon tissue sections;serum inflammatory factor tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),interleukin-1β(IL-1β)and interleukin-18(IL-18)were determined by enzyme linked immunosorbent assay(ELISA);myeloperoxidase(MPO)and superoxide dismutase(SOD)activities,nitric oxide(NO)and malondialdehyde(MDA)levels were detected by physiological and biochemical assay kits;protein expression levels of zonula occludens protein-1(ZO-1),occludin,nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3),and sirtuin 6(SIRT6)in mouse colon tissues were detected by western blot.[Results]Compared with normal group,DSS induced colonic inflammation,crypt loss,cuprocyte depletion and inflammatory infiltration;serum TNF-α,IL-6,IL-1β,and IL-18 inflammatory factor levels,MPO activity,NO and MDA levels were significantly increased(P<0.01,P<0.001),and SOD activity was significantly decreased(P<0.01).After treatment,the herbal enema formula significantly improved the DSS-induced colonic inflammation of crypt loss,cuprocyte depletion,and inflammatory infiltration.The levels of serum TNF-α,IL-6,IL-1βand IL-18 inflammatory factors,MPO activity and NO and MDA levels were significantly decreased(P<0.01,P<0.001),and SOD activity were significantly increased in medium dose and high dose groups(P<0.05).The protein expression levels of ZO-1,occludin and SIRT6 in the colonic tissues of m
作者
顾红
田玉雯
夏晶晶
马炯
GU Hong;TIAN Yuwen;XIA Jingjing(Jiangyin Hospital of Traditional Chinese Medicine,Jiangyin(214400),China;Nanjing University of Chinese Medicine)
出处
《浙江中医药大学学报》
CAS
2024年第11期1355-1362,共8页
Journal of Zhejiang Chinese Medical University
基金
江苏省中医药科技发展计划项目(YB2020046)
江阴市中医药学会科研项目(Z202202)。
