KKAy糖尿病脑病模型的探索及相关机制研究  

Exploration of KKAy Model with Diabetic Encephalopathy and Related Mechanism Study

作　　者：刘申贝 古小云 钟志江 赵军宁[2] 戴瑛[2] Liu Shenbei;Gu Xiaoyun;Zhong Zhijiang;Zhao Junning;Dai Ying(Pharmacy College of Southwest Medical University,Luzhou 646000;Sichuan Academy of Chinese Medicine Sciences,Sichuan Institute for Translational Chinese Medicine,Chengdu 610041;Zitong hospital of traditional Chinese medicine,Mianyang 622150)

机构地区：[1]西南医科大学药学院,泸州646000 [2]四川省中医药科学院,四川省中医药转化医学中心,成都610041 [3]绵阳市梓潼县中医院,绵阳622150

出　　处：《中药药理与临床》2020年第1期210-218,共9页Pharmacology and Clinics of Chinese Materia Medica

基　　金：国家科技重大专项"重大新药创制"(编号:2018ZX09731-008);四川省国际科技合作与交流计划项目(编号:2016HH0003)。

摘　　要：目的:针对糖尿病引起的认知障碍而目前研究缺少比较理想动物模型的难题,我们探索采用KKAy自发性2型糖尿病小鼠建立糖尿脑病模型,为糖尿病脑病机制以及新药筛选评价研究提供实验基础和依据。方法:采用转基因KKAy小鼠作为模型对照组,野生型C57BL/6小鼠作为正常对照组。通过对其体重血糖及糖耐量等指标的测定,判断是否出现了典型的2型糖尿病症状及胰岛素抵抗。通过行为学实验Morris水迷宫和小鼠自主活动实验验证认知功能损伤,通过病理染色检测其海马皮层神经元的改变、脑室皮层萎缩程度及眼微血管病变,采用免疫组化测定脑内BDNF的表达,生化法和ELISA检测其胆固醇(TC)、三酰甘油(TG)、高、低密度脂蛋白、胰岛素水平及炎症因子水平变化,Western Blot测定PI3K/AKT通路及Tau蛋白磷酸化表达变化程度。结果:与正常对照组相比,自发性KKAy小鼠血糖明显升高并维持高糖状态,体重明显增加,自主活动程度显著下降(P <0. 05或P <0. 01)。生化法结果显示,与正常对照组相比,模型对照组小鼠TC、TG、LDL-C水平显著升高,GSP含量显著增加,血清胰岛素增加,胰岛素抵抗指数增加(P <0. 05或P <0. 01)。Morris水迷宫结果发现,与正常对照组相比,模型对照组通过高脂饮食诱导至28周龄后出现认知功能显著下降,通过病理组织染色观察发现模型对照组小鼠脑室有明显扩张趋势,大脑皮层变薄萎缩,大量神经元细胞固缩,排列紊乱,海马椎体细胞层结构清晰,较多神经元细胞固缩。免疫组化结果发现,与正常对照组相比,模型对照组脑中BDNF表达明显减少。眼视网膜PAS染色发现,模型对照组视网膜血管出现明显损伤,可能诱导脑微血管病变。ELISA结果显示,与正常对照组相比,模型对照组TNF-α水平显著升高,IL-6和CRP水平有明显升高趋势。WB结果发现,模型对照组中海马和皮层组织中GSK-3β磷酸化表达�Objective: Due to the lack of an ideal animal model with cognitive impairment caused by diabetes in current research,we explored the establishment of diabetic encephalopathy model by using KKAy type 2 diabetic mice to provide experimental basis and basis for the mechanism of diabetic encephalopathy and the evaluation of new drug screening. Methods: Transgenic KKAy mice were used as the model,and wild type C57 BL/6 mice were used as the control. KKAy mice were fed on high fat diets. By measuring the indexes such as blood sugarweight and glucose tolerance,it is judged whether it had the typical symptoms of type 2 diabetes and insulin resistance. Morris water maze and mouse locomotor activity test were performed to verify whether mice had the cognitive impairment. Pathological staining was used to detect changes of hippocampal and cortex neurons,the cerebral atrophy,and ocular microvascular disease. Immunohistochemistry was used to determine the expression of BDNF in the brain. The changes of TC、TG,high density lipoprotein、low density lipoprotein、insulin levels and inflammatory factors were determined by biochemical methods and ELISA. Western Blot was used to determine protein expressions in PI3 K/AKT pathway and Tau protein phosphorylation. Results: In KKAy mice,the blood sugar significantly increased and maintained a high level,the body weights significantly increased,and the locomotor activity significantly decreased. Significant cognitive decline occurred after 28 weeks old. Biochemical test results showed that the levels of TC、TG、and LDL-C were significantly increased,the content of the glycated serum protein was significantly increased,the serum insulin was increased,and the insulin resistance index was increased. Morris water maze results showed that KKAy mice showed significant cognitive decline after 28 weeks of age. Pathological tissue staining revealed that the ventricles of the mice in the model group significantly were enlarged,the cerebral cortex became thin and atrophied,a large number of ne

关 键 词：糖尿病脑病 KKAy 认知障碍 发病机制 炎症 

分 类 号：R587.2[医药卫生—内分泌] R747.9[医药卫生—内科学] R-332[医药卫生—临床医学]