四神丸及拆方对大鼠实验性溃疡性结肠炎的影响  

Effect of Sishen Pill(四神丸)and Disassembling Prescription on Experimental Ulcerative Colitis in Rats

作　　者：于伟莉[1,2] 王炎 程杰 牛丽辉[1,2] 陈贵银 安福丽[1,2] 刘姣 YU Weili;WANG Yan;CHENG Jie;NIU Lihui;CHEN Guiyin;AN Fuli;LIU Jiao(Department of Pharmacy,Hebei Traditional Chinese Medical Hospital,Shijiazhuang 050021;Hebei Institute of Traditional Chinese Medical Preparation Industry Technology,Shijiazhuang 050021;College of Pharmacy,Hebei University of Chinese Medicine,Shijiazhuang 050200)

机构地区：[1]河北省中医院药学部,石家庄050021 [2]河北省中药制剂产业研究院,石家庄050021 [3]河北中医药大学药学院,石家庄050200

出　　处：《中药药理与临床》2024年第3期16-22,共7页Pharmacology and Clinics of Chinese Materia Medica

基　　金：河北省中医药管理局科研计划项目(编号:2022071)。

摘　　要：目的:观察四神丸、二神丸及五味子散对2,4,6-三硝基苯磺酸(2,4,6-trinitro-Benzenesulfonicacid,TNBS)诱导的溃疡性结肠炎大鼠(Ulcerative colitis,UC)的治疗作用,并从自噬的角度探讨其中机制及配伍意义。方法:采用TNBS灌肠法复制UC大鼠模型,将造模成功的大鼠随机分为模型对照组、美沙拉嗪0.36 g/kg组、四神丸3.4 g/kg组、二神丸3.2 g/kg组、五味子散3.2 g/kg组,每组10只;另设正常对照组10只。每日灌胃给予相应药物或蒸馏水,连续7 d。每日对大鼠的一般情况进行疾病活跃指数评分(Disease Activity Index,DAI);HE染色观察结肠组织病理变化;ELISA法检测结肠组织中白细胞介素(Interleukin,IL)-1β和肿瘤坏死因子(Tumor Necrosis Factor,TNF)-α含量;荧光Tunel法检测结肠下层黏膜组织中细胞凋亡情况;电镜检测结肠下层组织细胞中自噬小体;Western Blot法检测结肠程序性死亡受体-1(Beclin)-1、LC3Ⅱ/Ⅰ蛋白表达。结果:与正常对照组相比,模型对照组大鼠DAI评分、结肠组织中IL-1β和TNF-α含量均显著升高(P<0.01),结肠黏膜下层组织增生明显,细胞内未见典型自噬结构,结肠组织黏膜下层细胞凋亡率无明显变化,结肠LC3Ⅱ/Ⅰ和Beclin-1的蛋白表达明显下调(P<0.05或P<0.01);与模型对照组相比,四神丸3.4 g/kg组、二神丸3.2 g/kg组及五味子散3.2 g/kg组大鼠的一般情况明显改善,大鼠DAI评分显著降低(P<0.01),结肠黏膜下层组织增生减少,自噬小体数量增加;四神丸3.4 g/kg组及二神丸3.2 g/kg组大鼠结肠组织中IL-1β和TNF-α含量显著降低,结肠组织黏膜下层细胞凋亡显著增加(P<0.01),LC3Ⅱ/Ⅰ和Beclin-1的蛋白表达明显上调(P<0.05或P<0.01);与二神丸3.2 g/kg组和五味子散3.2 g/kg组比较,四神丸3.4 g/kg组UC大鼠的一般情况、组织病理和细胞超微结构病理变化改善更为显著,黏膜下层组织细胞凋亡率显著升高(P<0.01),Beclin-1的蛋白表达明显上调(P<0.05);与五味子散3.2Objective:To investigate the therapeutic effects of Sishen(四神)Pill,Ershen(二神)Pill,and Wuweizi(五味子)Powder on ulcerative colitis(UC)induced by 2,4,6-trinitro-benzenesulfonicacid(TNBS)in rats.The mechanism and compatibility significance were discussed from the perspective of autophagy.Methods:UC rat models were established by TNBS enema.The successfully modeled rats were randomly divided into a model group,mesalazine group(0.36 g/kg),Sishen Pill group(3.4 g/kg),Ershen pill group(3.2 g/kg),and Wuweizi Powder group(3.2 g/kg),with 10 rats in each group,and another 10 rats were selected as the control group.Corresponding drugs or distiled water were administered daily by gavage for seven days.The general condition of rats in each group was scored daily by disease activity index(DAI).Hematoxylin-Eosin(HE)staining was used to observe the pathological changes in colon tissue.Enzyme linked immunosorbent assay(ELISA)was used to detect the content of interleukin(IL)-1βand tumor necrosis factor(TNF)-αin colon tissue.The fluorescence TdT-mediated dUTP nick-end labeling(Tunel)method was used to detect the apoptosis of cells in colon submucosal tissue.Electron microscopy was used to detect autophagosomes in colon submucosal tissue cells.The protein expression levels of Beclin-1 and LC3 IV/I were detected by Western Blot.Results:Compared with the control group,the DAI score and the contents of IL-1βand TNF-αin colon tissue were significantly increased in the model control group(P<0.01).The proliferation of colon submucosal tissue was obvious,and no typical autophagy structure was observed in cells.The apoptosis rate of colon submucosal cells was not significantly changed(P>0.05).The protein expressions of LC3 I/I and Beclin-1 were significantly down-regulated(P<0.05 or P<0.01).Compared with the model control group,the general condition of the rats was improved in the Sishen Pill group(3.4 g/kg),Ershen Pill group(3.2 g/kg),and Wuweizi Powder group(3.2 g/kg),and the score of DAI was significantly reduced(P<0.01).Th

关 键 词：四神丸 二神丸 五味子散 自噬 凋亡 

分 类 号：R285.5[医药卫生—中药学]