【作者】 李春花；

【导师】 鞠玉琳；

【作者基本信息】 延边大学 ， 基础兽医学， 2008， 硕士

【摘要】 为了筛选出对大肠杆菌抑菌效果最好的复方中药,本试验采用药敏纸片法测定单味中药和复方中药的抑菌作用,以筛选出抑菌效果最好的复方中药制剂。通过试管二倍稀释法和平板稀释法测定复方中药制剂对大肠杆菌的MIC,并用药敏纸片法测定抑菌直径来判断体外抑菌效果,同时建立小鼠大肠杆菌腹泻模型,以腹泻抑制率来判断体内抑菌效果。本试验还从小鼠肠系膜毛细血管炎性渗出试验、小肠推进试验、抗毒素作用试验和牛小肠上皮细胞粘附试验等四个方面初步探讨了复方中药制剂抗大肠杆菌性腹泻作用的原因。结果:通过药敏纸片法对抑菌直径的测定可知金银花、黄连和连翘较其它七种单味中药对大肠杆菌的抑菌效果好,再将金银花、黄连和连翘按一定比例配伍组方得出复方二的抑菌作用最强,并对复方二添加辅药命名为中药复方“连黄”。经MIC的测定结果可知,试管二倍稀释法和平板稀释法结果一致,中药复方“连黄”对大肠杆菌的MIC都为3.91mg/ml,药敏纸片法测得抑菌直陉为18mm。体内抑菌试验结果显示,高、中、低剂量组对感染大肠杆菌小鼠的腹泻抑制率分别为58%、41%和24%,说明中药复方“连黄”抑制小鼠腹泻的作用与药物剂量呈正相关。小鼠肠系膜毛细血管炎性渗出试验结果表明,高、中剂量组小鼠腹水中伊文思蓝含量均极显著低于阴性对照组(P＜0.01),低剂量组与阴性对照组差异不显著(P＞0.05),高、中、低剂量组小鼠肠系膜毛细血管炎性渗出抑制率分别为53%、46%和28%。对小肠运动影响的试验结果表明,高、中剂量组小肠推进率均极显著低于阴性对照组(P＜0.01),低剂量组与阴性对照组差异不显著(P＞0.05),高、中、低剂量组对小肠运动的抑制率分别为45%、37%和20%。抗毒素作用试验结果表明,高、中剂量组肠袢的液体蓄积量极显著(P＜0.01)和显著(P＜0.05)低于阴性对照组,低剂量组与阴性对照组差异不显著(P＞0.05),说明药物剂量越大抗毒素作用越强。牛小肠上皮细胞粘附试验结果表明,高、中、低剂量组每个上皮细胞上粘附大肠杆菌的平均数分别为4、10和15个。结论:通过单味中药和复方中药的筛选确定了中药复方“连黄”;中药复方“连黄”能有效的抑制大肠杆菌的生长和繁殖;中药复方“连黄”抗大肠杆菌性腹泻作用的原因是通过抑制小鼠肠系膜毛细血管炎性渗出、抑制小肠运动、抗毒素作用和抑制大肠杆菌粘附于牛小肠上皮细胞而实现的。更多还原

【Abstract】 To find out the best Chinese traditional compound medicine which can inhibit E.coli strongest,the test uses paper disk method to determine antibacterial activity of single Chinese herb and the Chinese traditional compound medicine, to find out the best Chinese traditional compound medicine.The MIC of Chinese traditional compound medicine against E.coli is determined by tube double dilution and plate dilution method,the diameter of bacteriostasis ring is also determined by paper disk method to judge antibacterial activity in vitro,establish the mice model of E.coli diarrhea at the same time,to judge the antibacterial activity by inhibition ratio of diarrhea.The test also discuss the mechanism of Chinese traditional compound medicine against diarrhea of E.coli from flowing five accepts: Inflammatory effusion test of mice blood capillary、the facilitation test of small intestine、the test of antitoxin、adhesion test of small intestine epithelium cell of cattle and so on.Results:Antibacterial effects of Honeysuckle、Rhizome coptidis and Forsythia fruit are better than other seven single Chinese herb by paper disk method to determine the diameter of bacteriostasis ring,and Chinese traditional compound medicine two that is made up of Honeysuckle、Rhizome coptidis and Forsythia fruit has the strongest antibacterial action,which is named Chinese traditional compound medicine "LH" at the same time added auxiliary medicine.The results of tube double dilution and plate dilution method are coincidence from the result of MIC,determined MIC of "LH" against E.coli reaches 3.91mg/ml,the diameter of bacteriostasis ring is 18mm by paper disk method.The results of the bacteriostatic test in vitro shows:Inhibition rates of Diarrhea in mice infected with E.coli are that high-dose group is 58%,middle-dose group is 41%,low-dose group is 24%,which shows that the role of "LH" to inhibit mice diarrhea is positive correlation to the drug dose.The result of inflammatory effusion test in mice blood capillary shows:The content of Evans blue of high and middle-dose drug group in mice ascites is significantly lower than the negative control group (P＜0.01),low-dose group compare with the negative control group,there was no significant difference(P＞0.05).The ratioes of inflammatory effusion test in mice blood capillary of high-dose group,middle-dose group and low-dose are respectively 53%、46%、28%.The result of intestinal motility shows:intestinal facilitation ratio of high and middle-dose group are significantly lower than the negative control group(P＜0.01),there is no significant difference between the low-dose group and the negative control group(P＞0.05),the inhibition ratioes of intestinal motility are respectively 45%、37%、20%.The result of antitoxin shows:The number of ansa interstinalis fluid accumulation of high middle-dose group are significant(P＜0.01) and remarkable(P＜0.05) lower than the negative control group,there is no significant difference(P＞0.05) between the low-dose group and the negative control group,which shows drug doses is greater and the role of antitoxin is greater.The result of endothelial cell adhesion test shows: Average number of E.coli adhesing intestinal epithelial cell in high、middle and low-dose group is respectively 4、10、15.Conclusion:Chinese traditional compound medicine "LH" is determined by filtrating of single Chinese herb and the Chinese traditional compound medicine; "LH" can inhibit the growth and multiplication of E.coli;Mechanism of "LH" inhibiting diarrhea of E.coli is come true by inhibiting inflammatory effusion of blood capillary in mice mesentery,intestinal movement,antitoxin and E.coli to adhesie intestinal epithelial cell.更多还原