【作者】 高丽；

【导师】 李琳琳； 王烨；

【作者基本信息】 新疆医科大学 ， 药理学， 2020， 硕士

【摘要】 目的:本研究基于F.prausnitzii菌的抗炎机制,探究二甲双胍对db/db小鼠SCFAs、肠粘膜屏障作用及炎症因子的影响。方法:(1)将小鼠分为db/m+Na Cl、db/db+Na Cl、db/db+met组。于0,4,6,8周测定小鼠FBG、TC、TG水平。(2)采用ELISA测定小鼠IL-10,IL-6和胰岛素水平。(3)采用RT-PCR技术动态监测小鼠粪样中F.prausnitzii菌水平。(4)采用HE染色和油红O染色对小鼠相关组织切片进行染色。(5)采用WB检测小鼠结肠组织中claudin-1、claudin-2,zo-1、occlaudin、NF-кB蛋白表达。(6)采用GC测定小鼠粪样中SCFAs水平。结果:(1)表型指标结果:与db/db+Na Cl组小鼠相比,db/db+met组FBG、TC、TG和体重显著降低。(2)ELISA结果:与db/db+Na Cl组相比,db/db+met组IL-6水平降低;IL-10水平升高;空腹胰岛素水平降低。(3)RT-PCR实验结果:与db/db+Na Cl组相比,db/db+met组粪样中F.P菌水平升高。(4)染色结果:与db/db+Na Cl组相比,db/db+met组肠组织病理情况改善,肝脏脂肪减少。(5)WB结果:与db/db+Na Cl组相比,db/db+met结肠组织中claudin-1、zo-1、occlaudin蛋白表达增加,claudin-2蛋白减少,NF-кB蛋白有减少趋势。(8)GC法检测结果:与db/db+Na Cl组相比,db/db+met组乙酸,丁酸水平升高,丙酸水平无明显变化。结论:二甲双胍抗炎,保护肠粘膜屏障及产升高丁酸的作用均与F.prausnitzii菌功能相一致,提示F.prausnitzii菌在二甲双胍改善db/db小鼠糖脂代谢紊乱中发挥了重要作用。更多还原

【Abstract】 Objective:This study investigated the effects of metformin on SCFAs,intestinal mucosal barrier protection,and inflammatory factors in db/db mice based on the anti-inflammatory mechanism of F.prausnitzii.Methods:(1)Subjects:db/m+Na Cl group,db/db+Na Cl group,db/db+met group.FBG,TC,and TG levels were measured at 0,4,6 and 8 weeks after8 weeks of gavage.(2)Enzyme immunoassay was used to determine IL-10,IL-6,and insulin levels in serum of mice.(3)The levels of F.perusnitzii bacteria in fecal samples of mice at 0 to 8 weeks were determined by RT-PCR.(4)Colons and ileum sections were stained with HE staining and paraffin liver sections were stained with oil red O staining,and the pathological condition and fat content of the mice were examined.(5)The expression levels of claudin-1,claudin-2,zo-1,occlaudin and NF-кB in the colon tissues of mice were detected by western-blot.(6)The contents of acetic acid,propionic acid and butyric acid in 0-8-week mouse feces samples were determined by gas chromatography.Results:(1)Results of general biochemical indicators:compared with mice in the db/db+Na Cl group,fasting glucose level,triglyceride level,cholesterol level and body weight in the db/db+met group is significantly decreased,with statistically significant differences.(2)ELISA results:compared with mice in the db/db+Na Cl group,the serum level of pro-inflammatory factor IL-6 in the db/db+met group is decreased,with statistically significant difference.The level of anti-inflammatory factor IL-10 increases with statistically significant difference.Fasting insulin level is decreased,and the difference is statistically significant.(3)Rt-pcr results:Compared with mice in the db/db+Na Cl group,the level of F.prausnitzii bacteria in the feces of mice in the db/db+met group is significantly increased,with a statistically significant difference.(4)Pathological section results:compared with the mice in the db/db+Na Cl group,the colonic glands in the mice in the db/db+met group are arranged regularly,and the muscle layer is thickened.The structure of the ileum crypt is arranged in order and the muscular layer isthickened.Liver fat decreased significantly.(5)Western-blot results:compared with mice in the db/db+Na Cl group,claudin-1,zo-1 and occlaudin protein expressions in the colon tissues of db/db+met is increased,while claudin-2 protein is decreased,with statistically significant differences.The expression of inflammatory protein NF-кB is decreased,but the difference was not statistically significant.(8)GC test results: Compared with the db/db+Na Cl group,the acetic acid and butyric acid levels in the db/db + met group were significantly increased,and the difference was statistically significant.The propionic acid level was not significant Variety.Conclusion:The effects of metformin on anti-inflammatory,protecting the intestinal mucosal barrier and increasing the production of butyric acid are consistent with the functions of F.prausnitzii,suggesting that F.prausnitzii has played an important role in the improvement of glucose and lipid metabolism disorders in db/db mice.更多还原