| In order to investigate the prevention and cure mechanism of compound radix pulsatillae decoction (RP) for diarrhea, this test focuses on the effects of RP on the activitie of disaccharidase in intestinal mucosa in diarrheal mice induced with E. Coli. Method:one hundred and eighty BALB/c mice were randomly assigned into four groups:Prevention group, Treatment group, Self-healing group and Control group. Each mice was injected intraperitoneal with 2.6 billion E.coli O101 to induce diarrhea in all groups except the control group.The mice of Prevention group were fed with 1 mL RP once daily 7 days before the infection and last till the end of the test; The mice of Treatment group were fed with 1 mL RP once a day post infection;The mice of Self-healing group were fed with 1 mL physiological saline as the substitute once a day post infection; The mice of control group were treated with physiological saline as the substitute. The clinical symptoms were observed everyday.6 mice of each group were randomly selected for dissection on the every days post infection, the pathologic changes were observed. Posterior segment of duodenum was fixed immediately for making paraffin section. Histopathology changes were observed by light microscope. The mucosa of duodenum, jejunum and ileum were scraped, mashed and refrigerated respectively. The activities of lactase, maltase and sucrase of the mucosa homogenate were measured by the glucose oxidase method.The expression of the lactase gene of intestinal mucosa were measured by PCR method.The results as follow:Clinical symptoms:4h post infection the mice appeared depression, fatigue, chill, diarrhea and less feed intake.3d post infection the symptoms of prevention group and treatment group were decreased, the diarrhea were stopped. The symptoms of self-healing group were gradually decreased until 6d post infection. The mice of control group showed normally.Pathologic changes:the mice of prevention group, treatment group, and self-healing group appeared flatulence and hemorrhage in intestine severely, the liver, kidney, spleen enlarged, mesenteric lymph node were swelling, and intestine is full of yellow Loose stools, and intestinal tract was thinner to pull off. The mice of control group showed normally.3d post infection the symptoms of prevention group and treatment group were no hemorrhage, stool was grainy in rectum.Histopathological changes:the mice of prevention group,treatment group, and self-healing group appeared diarrhea after infected E. coli, the small intestine villi appeared partial ruptured, villous epithelial cell were swelled and necrosis partly, submucosa were congestion, edema and some inflammatory cell infiltration.The numbers of goblet cells were increased. The intestinal gland became degeneration obviously.4d post infection the symptoms of prevention group and treatment group were gradually decreased. Compared with the self-healing, the number of goblet cells did not increased significantly in treatment group, and other lesions consistent with the self-healing group. The number of goblet cells and submucosal edema had no significant changes in prevention group, Control group mice had the normal intestinal organizational structure.Intestinal mucosa disaccharidase changes:①Changes of lactase activity:it shows that duodenum and jejunum lactase activity is much higher than ileum after infected E. coli.The lactase activity of prevention group, treatment group and self-healing group were significantly lower than that of control group(P<0.01). The lactase activity of self-healing group, prevention group and treatment group had no differences one day post the infection (P>0.05).2d past infection the lactase activity of prevention and treatment groups was significantly higher than that of the self-healing group(P<0.01); 6d post infection, the lactase activity of prevention group, treatment group and control group were no significant difference but they were all significantly higher than that of the self-healing group(P<0.01);②Changes of invertase activity:The invertase activity of prevention group, treatment group and self-healing group were significantly lower than that of control group (P<0.01). The invertase activity of self-healing group, prevention group and treatment group had no differences one day post the infection (P>0.05).2d past infection the invertase activity of prevention and treatment groups was significantly higher than that of the self-healing group(P<0.01); 6d post infection, the invertase activity of prevention group, treatment group and control group were no significant difference but they were all significantly higher than that of the self-healing group(P＜0.01);③Changes of maltase activity:The maltase activity of prevention group, treatment group and self-healing group were significantly lower than that of control group(P<0.01), and they had no differences one day post the infection(P>0.05).7d post infection, the maltase activity of prevention group, treatment group and control group were no significant difference but they were all significantly higher than that of the self-healing group(P<0.01);The expression of mucosal lactase gene display:The expression of Gene Lac of prevention group, treatment group and self-healing group were significant lower than that of control group (P<0.01). The expression of Gene Lac of self-healing group, prevention group and treatment group had no differences one day post the infection (P>0.05). (P>0.05); 5d post infection,the expression of Gene Lac of prevention group, treatment group and control group were no significant difference but they were all significantly higher than that of the self-healing group(P<0.01);Conclusion:According to clinical symptoms,pathologic changes, histopathology changes and the activity of disaccharidase of the mice in each group, compound radix pulsatilla have significant preventive effects for diarrhea in mice, it can promote activity recovery of intestinal disaccharidase and the gene expression of intestinal lactase. The results showed that compound radix pulsatilla had the significantly preventive effects for diarrheal mice, and the preventive effect of compound radix pulsatilla may be closely related to regulat the activity of mucosal disaccharidase in intestinal. |
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