摘要
目的:围绕肝细胞线粒体的形态、产能和线粒体未折叠蛋白反应(mitochondrial unfolded protein response,UPR^(mt)),研究半夏泻心汤治疗2型糖尿病的可能机制。方法:选取若干只SPF级雄性SD大鼠,适应性喂养1周后,随机选取8只为正常组给予普通饲料喂养,其余给予高脂高糖饲料喂养4周后,腹腔注射链脲佐菌素(STZ)诱导T2DM模型;将造模成功的大鼠24只,随机分为模型组、半夏泻心汤组(中药组)、二甲双胍(MET)组,每组8只。采用血糖仪检测空腹血糖(fasting plasma glucose,FPG);比色法和分光光度计法测定肝组织ATP和反应性氧簇(reactive oxygen spieces,ROS)含量;透射电镜观察肝细胞线粒体超微结构;Western blot法检测UPR^(mt)相关蛋白表达。结果:模型组、MET组和中药组的体质量均低于正常组,FPG均高于正常组,差异均有统计学意义(P<0.05)。MET组和中药组的FPG均低于模型组,差异均有统计学意义(P<0.05)。模型组和MET组的ATP均低于正常组,且中药组高于模型组,差异均有统计学意义(P<0.05)。模型组和中药组的ROS均高于正常组,但MET组和中药组均低于模型组,差异均有统计学意义(P<0.05)。模型组线粒体肿胀,电子密度下降;MET组和中药组线粒体形态基本正常,但也有电子密度下降情况。中药组LonP表达高于正常组、模型组和MET组,差异均有统计学意义(P<0.05)。模型组ClpX表达低于正常组,MET组、中药组均高于模型组,差异均有统计学意义(P<0.05)。模型组ClpP表达低于正常组,MET组、中药组均高于模型组,且中药组高于MET组,差异均有统计学意义(P<0.05)。结论:T2DM发生时肝细胞UPR^(mt)损伤,而半夏泻心汤可能通过提升UPR^(mt)而保护肝细胞线粒体,从而有效干预2型糖尿病。
Objective:To study the possible mechanim of Banxia Xiexin Decoction treating type 2 diabetes mellitus(T2DM)based on mitochondrial morphology,generating energy and mitochondrial unfolded protein response(UPR^(mt))in liver cells.Method:A number of SPF male SD rats were selected,after 1 week of adaptive feeding,8 rats were randomly selected to be fed with normal diet,and the rest were fed with high fat and high sugar diet for 4 weeks,then Streptozotocin(STZ)was intraperitoneally injected to induce T2DM model.24 rats were randomly divided into model group,Banxia Xiexin Decoction group(Chinese medicine group)and Metformin(MET)group,with 8 rats in each group.Glucomere was used to detect rat fasting plasma glucose(FPG);colorimetry and photometer were employed to measure ATP and reactive oxygen spieces(ROS)levels respectively;transmission electron microscopy was applied to observe mitochondrial ultrastructures in liver cells;Western blot was chosen to detect expressions of proteins related UPR^(mt).Result:Body mass of the model group,MET group and Chinese medicine group were lower than that of the normal group;FPG of the model group,MET group and Chinese medicine group were higher than that of the normal group,the differences were statistically significant(P<0.05).The FPG of the MET group and Chinese medicine group were lower than that of the model group,the differences were statistically significant(P<0.05).ATP of the model group and MET group were lower than that of the normal group,but that of the Chinese medicine group was higher than that of the model group,the differences were statistically significant(P<0.05).ROS of the model group and Chinese medicine group were higher than that of the normal group,but MET of the model group and Chinese medicine group were lower than that of the model group,the differences were statistically significant(P<0.05).Mitochondrial swelling and electron density decreased in model group.The morphology of mitochondria in the MET group and Chinese medicine group were basically normal,but electron density also decreased.LonP expression of the Chinese medicine group was higher than those of the normal group,the model group and the MET group,the differences were statistically significant(P<0.05).The expression of ClpX of the model group was lower than that of the normal group,while those of the MET group and the Chinese medicine group were higher than that of the model group,the differences were statistically significant(P<0.05).The expression of ClpP of the model group was lower than that of the normal group,but those of the MET group and Chinese medicine group were higher than that of the model group,and Chinese medicine group was higher than that of the MET group,the differences were statistically significant(P<0.05).Conclusion:UPR^(mt)is damaged in T2DM liver cells,and Banxia Xiexin decoction might protect liver cell mitochondria via enhancing UPR^(mt)to treat T2DM effectively.
作者
马丽娜
马丹
许欣竹
王艺璇
刘文俊
段志园
单德红
MA Li’na;MA Dan;XU Xinzhu;WANG Yixuan;LIU Wenjun;DUAN Zhiyuan;SHAN Dehong(Integrated Chinese and Western Medicine College of Liaoning University of Traditional Chinese Medicine,Shenyang 110847,China;不详)
出处
《中国医学创新》
CAS
2021年第32期1-6,共6页
Medical Innovation of China
基金
国家自然基金面上项目(81673851)。
关键词
2型糖尿病
肝细胞
线粒体未折叠蛋白反应
半夏泻心汤
二甲双胍
Type 2 diabetes mellitus
Liver cell
Mitochondrial unfolded protein response
Banxia Xiexin Decoction
Metformin
