摘要
目的:从调控自噬清除过氧化物角度,探究人参败毒散修复溃疡性结肠炎(UC)肠黏膜的机制。方法:自由饮用3.0%葡聚糖硫酸钠(DSS)溶液诱导UC小鼠模型,雄性C57BL/6J小鼠共60只,随机分为正常组、模型组、阳性药美沙拉嗪组(0.3 g·kg^(-1))、人参败毒散高、中、低剂量组(12.35、8.22、4.11 g·kg^(-1)),每组10只,连续给药灌胃7 d。选用苏木素-伊红(HE)染色观察结肠组织病变,阿利新蓝-过碘酸雪夫氏(PAS/AB)染色观察杯状细胞改变。免疫荧光法检测结肠组织活性氧自由基(ROS)荧光表达;生化法检测超氧化物歧化酶(SOD)活性、丙二醛(MDA)含量。蛋白免疫印迹法(Western blot)检测结肠组织细胞增殖核抗原(PCNA)、微管相关蛋白1轻链3(LC3)、富含亮氨酸重复序列G蛋白偶联受体5(LGR5)、p62的表达水平。结果:与正常组比较,UC模型组结肠HE染色见黏膜上皮结构大量缺失,肠腺破坏,炎性细胞大量浸润,结肠病理损伤评分(TDI)明显上调(P<0.05),杯状细胞大量丢失,SOD活性及LC3Ⅱ/Ⅰ、PCNA下调(P<0.05),而p62、MDA、ROS、LGR5水平上调(P<0.05);与模型组比较,不同剂量的人参败毒散组结肠TDI评分均明显下降(P<0.05),PAS/AB染色见大量新生的杯状细胞。PCNA、LGR5、SOD活性及LC3Ⅱ/Ⅰ明显上升(P<0.05),而p62、MDA、ROS含量明显降低(P<0.05),以人参败毒散低剂量组最佳(P<0.05)。结论:人参败毒散可促肠上皮修复,其机制可能与启动肠道自噬,减轻氧化应激损伤,促使肠干细胞增殖、分化有关。
Objective:To explore the mechanism of Renshen Baidusan in repairing intestinal mucosa in ulcerative colitis(UC)by regulating autophagy to scavenge peroxides.Method:The mouse model of UC was induced by free drinking of 3.0%dextran sulphate sodium(DSS)solution.Sixty male C57BL/6J mice were randomized into normal,model,mesalazine(0.3 g·kg^(-1)),and high-,medium-,and low-dose(12.35,8.22,4.11 g·kg^(-1),respectively)Renshen Baidusan groups(n=10).The mice were administrated with corresponding drugs by gavage for 7 consecutive days.The colon tissue was stained with hematoxylin-eosin(HE)to reveal the pathological changes,and Alcian blue-Periodic acid Scheff(PAS/AB)staining was employed to observe the goblet cell changes.The fluorescence expression of reactive oxygen species(ROS)in the colon tissue was detected by the immunofluorescence assay.The activity of superoxide dismutase(SOD)and the content of malondialdehyde(MDA)were measured by the biochemical methods.Western blot was employed to determine the expression levels of proliferating cell nuclear antigen(PCNA),microtubule-associated protein 1 light chain 3(LC3),leucine-rich repeat-containing G protein-coupled receptor 5(LGR5),and p62.Result:Destroyed mucosal epithelial structure,intestinal gland destruction,loss of goblet cells,and massive infiltration of inflammatory cells appeared in the model group.Compared with the normal group,the model group showed increased tissue damage injury(TDI)score of the colon tissue,decreased SOD activity and LC3Ⅱ/Ⅰ,PCNA value,and elevated levels of p62,MDA,ROS,and LGR5(P<0.05).Compared with the model group,different doses of Renshen Baidusan decreased the TDI score,promoted the generation of new goblet cells,elevated the levels of PCNA,LGR5,SOD,and LC3Ⅱ/Ⅰ,and lowered the levels of p62,MDA,and ROS(P<0.05).Moreover,the low dose group showed the best performance(P<0.05).Conclusion:Renshen Baidusan can promote intestinal epithelial repair by activating intestinal autophagy,alleviating oxidative stress,and promoting intestinal stem
作者
熊珮宇
陈旭
张薇
刘俊宇
刘兴隆
王一童
贾波
XIONG Peiyu;CHEN Xu;ZHANG Wei;LIU Junyu;LIU Xinglong;WANG Yitong;JIA Bo(School of Basic Medical Sciences,Chengdu University of Traditional Chinese Medicine,Chengdu 610075,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2023年第19期34-41,共8页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金面上项目(82074327)。
关键词
人参败毒散
溃疡性结肠炎
自噬
氧化应激
肠干细胞
Renshen Baidusan
ulcerative colitis
autophagy
oxidative stress
intestinal stem cells
