Effect of Sishen Pill(四神丸)and Disassembling Prescription on Experimental Ulcerative Colitis in Rats
Objective:To investigate the therapeutic effects of Sishen(四神)Pill,Ershen(二神)Pill,and Wuweizi(五味子)Powder on ulcera-tive colitis(UC)induced by 2,4,6-trinitro-benzenesulfonicacid(TNBS)in rats.The mechanism and compatibility significance were dis-cussed 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 Wu-weizi 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 distilled water were administered daily by gavage for seven days.The general condition of rats in each group was scored daily by disease activ-ity index(DAI).Hematoxylin-Eosin(HE)staining was used to observe the pathological changes in colon tissue.Enzyme linked immunosor-bent assay(ELISA)was used to detect the content of interleukin(IL)-1β and tumor necrosis factor(TNF)-α in colon tissue.The fluores-cence 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 Ⅱ/Ⅰ were detected by Western Blot.Results:Compared with the control group,the DAI score and the contents of IL-1β and TNF-α in colon tissue were significantly increased in the model control group(P＜0.01).The proliferation of colon submucosal tissue was obvious,and no typical autoph-agy structure was observed in cells.The apoptosis rate of colon submucosal cells was not significantly changed(P＞0.05).The protein ex-pressions of LC3Ⅱ/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).The proliferation of colon submucosa tissue was decreased,and the number of autophagosomes was increased.The contents of IL-1β and TNF-α in colon tissue in Sishen Pill group(3.4 g/kg)and Ershen Pill group(3.2 g/kg)were significantly decreased(P＜0.01),and the apoptosis of colon submucosal cells was significantly increased(P＜0.01).The protein expressions of LC3 Ⅱ/Ⅰ and Beclin-1 were significantly up-regulated(P＜0.05 or P＜0.01).Compared with the Ershen Pill group(3.2 g/kg)and Wuweizi Powder group(3.2 g/kg),the general condition,histopathology,and cell ultrastructure changes of UC rats in the Sishen Pill group(3.4 g/kg)were significantly improved,and the apoptosis rate of submucosal tissue cells was increased(P＜0.01).Beclin-1 protein expression level was up-regulated(P＜0.05).Compared with the Wuweizi Powder group(3.2 g/kg),the contents of IL-1β and TNF-α in colon tissue of rats in the Sishen Pill group(3.4 g/kg)were significantly decreased(P＜0.05 or P＜0.01).Conclu-sion:Sishen Pill and its disassembling prescription can promote autophagy,reduce the level of inflammatory factors,increase colon submuco-sal apoptosis,and alleviate UC symptoms.Moreover,the effect of Sishen Pill is better than Ershen Pill and Wuweizi Powder.
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