大黄蛰虫丸对AOM/DSS诱导结肠炎相关结直肠癌小鼠的抑制作用

Inhibitory Effects of Dahuang Zhechong Pill on AOM/DSS Induced Colitis-associated Cancer in Mice

目的:研究大黄蛰虫丸(DZP)对结肠炎相关结直肠癌(CAC)的影响及其可能机制。方法:以C57BL/6小鼠为研究对象,随机分为对照组、模型组,DZP低(2 g·kg^(-1))、高剂量组(4 g·kg^(-1))。除对照组外,其余各组采用AOM/DSS诱导小鼠CAC模型,并于造模中各组给予相应的药物灌胃干预。实验结束,比较各组小鼠死亡率,并以ELISA检测小鼠血清IL-1β和IL-18水平;苏木素-伊红(HE)染色观察结肠组织损伤情况;免疫荧光染色分析小鼠结肠Occludin和ZO-1的表达;免疫组化染色分析小鼠结肠ATG5、IL-1β和IL-18的表达水平;蛋白印迹检测小鼠结肠中LC3BⅠ/Ⅱ、SQSTM1及ATG5的蛋白表达水平。结果:经8周灌胃给药,对照组、模型组,DZP低、高剂量组的死亡率分别为0%、40.00%、20.00%、6.67%。与模型组相比,DZP能显著降低血清IL-1β和IL-18水平,提高小鼠肠道组织中Occludin和ZO-1的表达水平。同时,DZP能显著降低模型小鼠结肠组织中IL-1β、IL-18及SQSTM1的表达水平,提高模型小鼠肠道组织中的LC3BⅠ/Ⅱ和ATG5的蛋白表达水平。结论:DZP能显著降低CAC模型小鼠的炎症和死亡率;其作用机制可能与促进CAC模型小鼠肠道自噬改善肠道紧密连接相关。

Objective:To explore the impacts of Dahuang Zhechong Pills(DZP)on colitis-associated cancer(CAC)and their possible mechanism.Methods:C57BL/6 mice were randomly divided into control group,model group and DZP groups of low-dose(2 g·kg^(-1))and high-dose(4 g·kg^(-1)).Except the control group,the model of AOM/DSS induced CAC was established in mice of the other groups.During the modeling,mice in each group were treated with corresponding drugs by intragastric administration.At the end of experiment,the mortality rates in each group were compared,and the levels of IL-1βand IL-18 in serum were detected by ELISA.Hematoxylin and eosin(HE)staining was used to observe the injury of colon tissues.Immunofluorescence staining was used to observe the expression of Occludin and ZO-1 in colon tissues.The expression levels of ATG5,IL-1βand IL-18 in colon tissues were detected by immunohistochemistry.The protein expression of LC3BⅠ/Ⅱ,SQSTM1 and ATG5 in colon tissues were detected by Western blot.Results:The mortality rates were 0%in the control group,40.00%in the model group,20.00%in the low-dose DZP group and 6.67%in the high-dose DZP group.Compared with the model group,the treatment of DZP significantly reduced the levels of IL-1βand IL-18 in serum,and significantly increased the Occludin and ZO-1 expression in colon tissues.The expression levels of IL-1β,IL-18 and SQSTM1 were significantly decreased in colon tissues,but the levels of LC3BⅠ/Ⅱand ATG5 were highly increased.Conclusion:The DZP can significantly reduce the mortality rates and inflammation in CAC mice.The mechanism may be related to promoting intestinal autophagy and improving tight junction of intestinal.