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This test focuses on the mechanism of action of the Escherichia coli diarrhea Four Gentlemen Decoction control mice. Test methods: the establishment of animal by intraperitoneal injection of Escherichia coli diarrhea model, the test mice were divided into prevention group, the treatment group, self-healing group and the control group, attack bacteria mice each intraperitoneal injection of 260 million E. coli O101 induced diarrhea. Prevention group in advance 5d began with four gentlemen decoction orally orally 0.5mL until the end of the experiment, each morning and evening; treatment group: modeling with Sijunzitang decoction orally, morning and evening; self-healing group E. coli only modeling; blank control group, Sijunzitang decoction and E. coli broth suspension with saline instead. Daily observation of clinical symptoms, mice in each group necropsy three mice were observed record pathological change, using the conventional sectioning technique, electron microscopy techniques to observe pathological changes of mice in each group, and ultrastructural changes and Using immunohistochemistry and PCR methods to study the repair gene expression in mouse intestinal mucosa. The test results were as follows: clinical symptoms: self-healing group, the treatment group and the prevention group mice attack bacteria after 8h about symptoms of diarrhea, mice depression, fatigue, reduced food intake, coat against the legislation, abdominal breathing, rows of yellow dilute it, eyelid redness, discharge outflow, after 96 hours, and symptoms improved gradually restored; compared with the self-healing group of mice treated with a lower rate of diarrhea, mild clinical symptoms; significantly reduce the clinical symptoms of the prevention group, small the mouse diarrhea rates significantly reduce the recovery time is significantly shorter; blank control group of mice are no exception. The necropsy changes: attack bacteria, mouse liver, kidney, spleen enlargement, duodenum with scattered bleeding points, wall thinning, filled with thin yellow contents of the intestine, gall bladder full of bile, severe stomach cheerily. 120h, pathological change light; treatment group did not change significantly 96h after necropsy; prevent mice lesions significantly reduced, liver, kidney, spleen lesions disappeared, duodenal bleeding point reduction; blank control group necropsy not See exception. Histopathological changes: mouse attack bacteria, intestinal villi fracture villus epithelial cell swelling, partial necrosis, congestion and edema of the submucosa, a small amount of inflammatory cell infiltration, an increase in the number of goblet cells, midgut gland degeneration. 120h, the lesion has been reduced and gradually restored. Treatment group, the number of goblet cells did not change significantly, and pathological changes after 120h gradually reduce; prevention group, the number of goblet cells and submucosa congestion and edema did not change significantly, gradually restored after 96 hours compared with the self-healing group; intestinal blank control group organizational structure normal fluff neat, clear outline of the midgut gland. Ultrastructural changes: Scanning electron microscopy showed that attack bacteria, fluff swelling, abnormal morphology of goblet cells, the increase in the number of top the microvilli swelling shedding, sparse and arranged messy. 96h, villous architecture gradually restored; treatment group compared with self-healing lesions were lighter; prevention group ultrastructural lesions lighter microvilli, disorganized, and without shedding phenomenon gradually restored after 72 hours; blank control group, small intestinal villus height neat, evenly distributed dense microvilli neat goblet cell number, morphology and distribution were normal. TEM observations showed that after the attack the bacteria, the rough endoplasmic reticulum expansion of the formation of many small vacuoles, there is the phenomenon of exocytosis and vesicle. Epithelial microvilli derangement. Mitochondrial vacuolization, the crest and film disappear. Goblet cells contain a lot of sticky original particles; ultrastructural lesions improved and gradually restored after 96 hours; treatment group with the same set of basic, self-healing and prevention group ultrastructural lesions lighter; blank control group the intestinal absorption epithelial microvilli neat and normal mitochondrial structure and integrity of the organelle. The immunohistochemistry results: diarrhea early prevention group, the treatment group and the self-healing group than in the control group of PCNA of TGFβ1 expression was increased in the prevention group and the control group, the difference was significantly (P lt; 0.05). Mucosal cell repair gene expression test results showed that diarrhea early prevention group, the treatment group, and self-healing group the TGFβ1, EGFR expression compared with blank control group, a significant difference (P lt; 0.01), the prevention group, treatment group and self-healing group compared with a significant difference (P lt; 0.05). Conclusion: Prevention mice display the preventive effect Sijunzitang significant clinical symptoms and pathological changes, histopathological changes, as well as electron microscopy results. The prevention group mucosal repair gene of PCNA, EGFR, of TGFβ1 expression was significantly increased. The results show that the control effect of Sijunzitang E. coli diarrhea in mice may be related to regulation of the mucosal repair genes of PCNA, EGFR, of TGFβ1 expression is closely related.
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