目的 探究高同型半胱氨酸血症（hyperhomocysteinemia,HHcy）对血糖稳态的影响。方法 将C57BL/6小鼠随机分为对照组（CHOW）、模型组（HMD）两组,每组10只。CHOW组使用对照饲料（AIN-93G）饲养;HMD组使用高蛋氨酸（AIN-93G+2%）饲料饲养。HHcy模型构建成功后,对两组小鼠进行葡萄糖耐量实验（oral glucose tolerance test,OGTT）、胰岛素耐量实验（insulin tolerance test,ITT）和胰岛素分泌实验。饲养4月后处理取材,取小鼠血浆和胰腺组织,采用苏木素-伊红（hematoxylin-eosin,HE）染色观察小鼠胰腺病理学组织形态变化、免疫荧光染色技术检测胰腺胰岛素表达、酶联免疫吸附试验（enzyme linked immunosorbent assay,ELISA）检测胰腺胰岛素总含量。结果 HMD组小鼠蛋氨酸饮食1月后,与CHOW组相比,血浆中Hcy水平显著增高（P<0.0001）,成功构建HHcy模型;饲养4月后,与CHOW组小鼠相比,两组小鼠空腹血糖无明显差异,HMD组小鼠随机血糖显著升高（P<0.0001）、糖耐量受损（P<0.001）、胰岛素耐量受损（P<0.001）;与CHOW组小鼠相比,HMD组小鼠葡萄糖刺激下的胰岛素分泌减少（P<0.0001）;HE染色结果显示,两组小鼠胰腺组织形态无明显变化;小鼠胰腺胰岛素免疫荧光化学分析显示,与CHOW组小鼠相比,HMD组小鼠胰岛素荧光强度增强（P<0.001）;与CHOW组小鼠相比,HMD组小鼠胰腺胰岛素总含量显著上升,差异有统计学意义（P<0.0001）。结论 HHcy可以引起T2DM（type 2 diabetes mellitus,T2DM）的发生,同型半胱氨酸（homocysteine,Hcy）可能通过损伤胰岛素分泌功能影响血糖稳态。
Abstract
Objective To explore the effects of hyperhomocysteinemia (HHcy) on blood glucose homeostasis. Methods C57BL / 6 mice were randomly divided into two groups: control group (CHOW) and model group (HMD), 10 mice per group. The CHOW group was fed the AIN-93G diet, and the HMD group fed the AIN-93G+2% methionine diet. The oral glucose tolerance test (OGTT), insulin tolerance test (ITT), and insulin secretion experiment were performed. The morphological changes in the pancreas were observed by hematoxylin-eosin (HE) staining, pancreatic insulin expression by immunofluorescence staining, and total pancreatic insulin content by enzyme-linked immunosorbent assay (ELISA). Results Compared with the CHOW group, after 1 month of feeding, plasma Hcy was significantly increased in the HMD group (P<0.0001). After 4 months of feeding, there was no significant difference in fasting blood glucose between the two groups, but mice in the HMD group had significantly increased random blood glucose (P<0.0001), impaired glucose tolerance (P<0.001) and insulin tolerance (P<0.001); Compared with the CHOW group, the glucose-stimulated insulin secretion was reduced in the HMD group (P<0.0001). The results of HE staining showed that there was no significant change in the pancreatic morphology between the two groups. Immunofluorescence chemical analysis revealed enhanced insulin fluorescence intensity in the pancreas of the HMD group (P<0.001). Compared with the CHOW group, the total pancreatic insulin content increased significantly in the HMD group. The difference was statistically significant (P<0.0001). Conclusion HHcy can cause the development of type 2 diabetes mellitus (T2DM), and homocysteine may affect blood glucose homeostasis by damaging insulin secretion function.
关键词
同型半胱氨酸 /
同型半胱氨酸血症 /
血糖稳态 /
胰岛β细胞 /
胰岛素分泌
Key words
Hcy /
HHcy /
blood glucose homeostasis /
pancreatic β-cells /
insulin secretion
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基金
天津市科技计划项目（No.21JCZDJC01070）; 天津市卫健委天津市中医药管理局中西医结合项目（No.2021031）
