| Objective: Based on the theory of "intestinal fistula",a prospective experimental study on the treatment of ulcerative enteritis(UC)with coix seed,aconitum carmichaelii and patrinia powder and its mechanism of intervention on the FAS pathway of ulcerative colitis and the expression of Caspase-8 protein in related colon cancer were observed.Methods:Male C57BL/6J mice were randomly divided into blank control group,model control group,positive drug(mesalazine)control group,low,medium and high dose of traditional Chinese medicine(Coix seed,aconitum carmichaelii and patrinia powder)three groups.There were eight mice in each group.Except for the blank control group,the mice in the remaining groups were free to drink 3% DSS solution for five consecutive days.Then,from the sixth day,the model mice were intraperitoneally injected with 1,2-dimethylhydrazine NS solution at a dose of 40mg/kg.Once every other day,three times in total to replicate the mouse model of ulcerative colitis.From the eleventh day,mice in each group were given the corresponding drugs by intragastric administration according to the following dosage for seven consecutive days and once a day in the morning and once a day in the afternoon.Positive drug control group(mesalazine,40 mg/kg);Low,medium and high dose groups of traditional Chinese medicine(Coix seed,aconitum carmichaelii and patrinia powder,3.32 g crude drug/kg,6.63 g crude drug/kg,13.26 g crude drug/kg).The normal control group and the model control group were given double distilled water in the same volume by intragastric administration until the end of the experiment.Detection index: general conditions of mice were observed and DAI scores of mice in each group were calculated synchronously during the experiment.On the seventeenth day of the experiment,the neck removal method was used to kill the mice.The abdominal cavity was immediately dissected to take colon-related tissues and detection was performed:(1)HE staining and HI score;(2)Immunohistochemistry was used to detect the expression of Fas and Fasl proteins;(3)Western blot technology was used to determine the expression of Caspase8 protein.(4)RT-PCR technology was used to determine the expression of mi R-21(m RNA).Results:(1)For DAI score,the model control group was higher than the normal control group(P<0.05).After the intervention,DAI scores of mice in all drug groups were significantly reduced(P< 0.05).The results of HE staining and HI score showed that in the model control group,the colonic mucosa was erosive and ulcerated and most of the mucosa was defective.The glands were isolated and a large number of neutrophils were infiltrated with atypical hyperplasia.Compared with the model control group,the structure of the positive drug control group,the middle and high dose groups of traditional Chinese medicine were relatively complete.The infiltration of inflammatory cells decreased.Atypical hyperplasia was significantly improved(P<0.01).(2)Immunohistochemical results showed that the expression level of Fas/Fasl protein in blank control group was lower and that in model control group was significantly higher than that in blank control group(P<0.01).Compared with the model control group,this index in the positive drug control group,the middle and high dose groups of traditional Chinese medicine were significantly reduced(P<0.05 or P< 0.01).(3)Western Blot showed that the expression level of Caspase8 in blank control group was lower and that in model control group was significantly higher than that in blank control group(P< 0.01).Compared with the model control group,this index of all drug groups was significantly reduced(P<0.01).(4)RT-PCR results showed that the expression level of mi R21(m RNA)in the blank control group was lower and that in the model control group was significantly higher than that in the blank control group(P< 0.01).Compared with the model control group,this index of all drug groups was significantly reduced P < 0.01).Conclusion: In a certain dose range,Coix seed,aconitum carmichaelii and patrinia powder has obvious inhibitory effect on colon inflammatory reaction and colon tissue lesion of ulcerative colitis canceration mice induced by DSS-DMH.It can regulate the excessive apoptosis of intestinal epithelial cells and prevent the occurrence and development of colon cancer lesions caused by continuous inflammatory response injury of colon.Its effective way of developing colitis-related cancerous lesions may be through inhibiting the over-activation of Fas/Fas L transduction pathway,which decreases the damage degree of inflammatory reaction of this model mouse,inhibits the pathological occurrence and development process of "inflammation-atypical hyperplasia-cancerous lesions" and plays an effective role in preventing and treating ulcerative colitis-related colon cancer. |
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