不同造模方式诱导的糖尿病小鼠肝肾组织中脂质异位沉积情况的研究  

Comparative study on lipid deposition in liver and kidney tissues of diabetic mice induced by different modeling methods

作　　者：刘露 扈腊英 王贵芳 常雪冰 黄亚莉 宋铃榆 周宇霞 郭兵[1] LIU Lu;HU Laying;WANG Guifang;CHANG Xuebing;HUANG Yali;SONG Lingyu;ZHOU Yuxia;GUO Bing(Department of pathophysiology,Guizhou Medical University/Guizhou Key Laboratory of Pathogenesis and Drug Research of Common Chronic Diseases,Guiyang 550025,China)

机构地区：[1]贵州医科大学病理生理学教研室/贵州省常见慢性疾病发病机制及药物研究重点实验室,贵州贵阳550025

出　　处：《中国病理生理杂志》2023年第2期276-286,共11页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.82000741;No.32160207);贵州省科技厅计划项目(黔科合基础-ZK[2021]一般402号);中国博士后科学基金资助项目(No.2020M683374);贵州省教育厅青年科技人才成长项目(黔科KY字[2021]170号);贵州医科大学优秀青年人才计划(No.2021105)。

摘　　要：目的:观察比较高脂饮食(high-fat diet, HFD)、链脲佐菌素(streptozocin, STZ)联合HFD(HFD/STZ)、单侧肾脏切除术(unilateral nephrectomy, UNx)+HFD/STZ和db/db小鼠4种糖尿病模型小鼠肝肾组织中脂质异位沉积情况。方法:选取8周龄C57BL/6小鼠,随机分为正常饮食(normal chow diet, NCD)组(n=10)、HFD组(n=10)、HFD/STZ组(n=10)和UNx+HFD/STZ组(n=12)。NCD组给予NCD饲养;HFD组、HFD/STZ组和UNx+HFD/STZ组给予HFD饲养,诱导小鼠出现肥胖和胰岛素抵抗,且HFD/STZ组和UNx+HFD/STZ组给予55 mg/kg STZ腹腔注射,后以HFD饲养至40周龄。选取10周龄db/db小鼠和野生型小鼠各8只,NCD饲养至40周龄。记录分析小鼠血糖和体重变化;收集血液样本测定甘油三酯(triglyceride, TG)含量;肝肾组织样本石蜡包埋后进行苏木精-伊红染色观察组织病理变化;免疫荧光染色和Western blot检测脂滴包被蛋白2的表达;油红O染色和尼罗红染色观察脂滴沉积情况。结果:HFD组小鼠体重显著升高(P<0.05),血糖水平和血清TG含量升高不显著(P>0.05);HFD/STZ组、Unx+HFD/STZ组及db/db组血糖水平和血清TG含量显著升高(P<0.05),且肝肾组织中存在大量脂质异位沉积,均能诱导小鼠发生糖尿病肝肾脏损伤,其中Unx+HFD/STZ组最严重;HFD组肾脏未见明显脂质沉积,肝脏存在大量脂质沉积。结论:Unx+HFD/STZ组小鼠的肝肾脂质沉积最严重,但存在死亡率较高、小鼠状态差等缺点。单纯HFD饮食的小鼠是研究非酒精性脂肪性肝病合适的模型。HFD/STZ和db/db模型小鼠出现糖尿病肾脏脂毒性且呈现糖尿病肾病的肾脏病理改变,类似于2型糖尿病患者,适用于研究糖尿病及其并发症。AIM: To observe the lipid deposition in liver and kidney tissues of diabetic mice. METHODS: Diabetes mellitus models were induced by high-fat diet(HFD), streptozocin(STZ) combined with HFD(HFD/STZ), unilateral nephrectomy(Unx)+HFD/STZ and db/db mice. Eight-week-old C57BL/6 mice were randomly divided into normal chow diet(NCD) group(n=10), HFD group(n=10), HFD/STZ group(n=10), and Unx+HFD/STZ group(n=12). The mice in NCD group were fed with NCD. The mice in HFD group, HFD/STZ group and Unx+HFD/STZ group were fed with HFD. The mice in HFD/STZ group and Unx+HFD/STZ group were injected with STZ(55 mg/kg) and then continuously fed with HFD until to the 40th week. Ten-week-old db/db and wild-type mice(n=8) were fed with NCD until to the 40th week. Blood glucose level, body weight and serum triglyceride(TG) level of each group were detected. Tissue sections of the liver and kidney were made via dehydration and paraffin embedding. After hematoxylin-eosin staining, the histopathological changes of the liver and kidney tissues were observed by microscopy. The expression of perilipin 2 was detected by immunofluorescence staining and Western blot. Lipid deposition was observed by oil red O staining and Nile red staining.RESULTS: In HFD group, the body weight of the mice was significantly increased(P<0. 05), but the level of blood glucose and the content of serum TG were not significantly increased(P>0. 05). In HFD/STZ group, Unx+HFD/STZ group and db/db group, both blood glucose level and serum TG content were significantly increased(P<0. 05), lipid deposition and tissue damage were observed in the liver and kidney tissues. The lipid deposition in the liver and kidney tissues was the most obvious in Unx+HFD/STZ group. In HFD group, lipid deposition was observed in the liver but not in the kidney.CONCLUSION: In the liver and kidney tissues, the lipid deposition is the most obvious in Unx+HFD/STZ group, but there are some shortcomings such as high mortality and poor state of mice. The HFD model is an ideal model to study nonalc

关 键 词：糖尿病 脂质沉积 脂毒性 链脲佐菌素 单侧肾脏切除术 

分 类 号：R587.2[医药卫生—内分泌] R363.2[医药卫生—内科学]