PDF(Pubmed)
Abstract：
OBJECTIVE: Vascular complications contribute significantly to the extensive morbidity and mortality rates observed in people with diabetes. Despite well known that the diabetic kidney and heart exhibit imbalanced angiogenesis, the mechanisms implicated in this angiogenic paradox remain unknown. In this study, we examined the angiogenic and metabolic gene expression profile (GEP) of endothelial cells (ECs) isolated from a mouse model with type1 diabetes mellitus (T1DM).
METHODS: ECs were isolated from kidneys and hearts of healthy and streptozocin (STZ)-treated mice. RNA was then extracted for molecular studies. GEP of 84 angiogenic and 84 AMP-activated Protein Kinase (AMPK)-dependent genes were examined by microarrays. Real time PCR confirmed the changes observed in significantly altered genes. Microvessel density (MVD) was analysed by immunohistochemistry, fibrosis was assessed by the Sirius red histological staining and connective tissue growth factor (CTGF) was quantified by ELISA.
RESULTS: The relative percentage of ECs and MVD were increased in the kidneys of T1DM animals whereas the opposite trend was observed in the hearts of diabetic mice. Accordingly, the majority of AMPK-associated genes were upregulated in kidneys and downregulated in hearts of these animals. Angiogenic GEP revealed significant differences in Tgfβ, Notch signaling and Timp2 in both diabetic organs. These findings were in agreement with the angiogenesis histological assays. Fibrosis was augmented in both organs in diabetic as compared to healthy animals.
CONCLUSIONS: Altogether, our findings indicate, for the first time, that T1DM heart and kidney ECs present opposite metabolic cues, which are accompanied by distinct angiogenic patterns. These findings enable the development of innovative organ-specific therapeutic strategies targeting diabetic-associated vascular disorders.
METHODS: ECs were isolated from kidneys and hearts of healthy and streptozocin (STZ)-treated mice. RNA was then extracted for molecular studies. GEP of 84 angiogenic and 84 AMP-activated Protein Kinase (AMPK)-dependent genes were examined by microarrays. Real time PCR confirmed the changes observed in significantly altered genes. Microvessel density (MVD) was analysed by immunohistochemistry, fibrosis was assessed by the Sirius red histological staining and connective tissue growth factor (CTGF) was quantified by ELISA.
RESULTS: The relative percentage of ECs and MVD were increased in the kidneys of T1DM animals whereas the opposite trend was observed in the hearts of diabetic mice. Accordingly, the majority of AMPK-associated genes were upregulated in kidneys and downregulated in hearts of these animals. Angiogenic GEP revealed significant differences in Tgfβ, Notch signaling and Timp2 in both diabetic organs. These findings were in agreement with the angiogenesis histological assays. Fibrosis was augmented in both organs in diabetic as compared to healthy animals.
CONCLUSIONS: Altogether, our findings indicate, for the first time, that T1DM heart and kidney ECs present opposite metabolic cues, which are accompanied by distinct angiogenic patterns. These findings enable the development of innovative organ-specific therapeutic strategies targeting diabetic-associated vascular disorders.
摘要：
目的：血管并发症对糖尿病患者的广泛发病率和死亡率有显著影响。尽管众所周知，糖尿病肾脏和心脏表现出不平衡的血管生成，这一血管生成悖论的相关机制尚不清楚.在这项研究中,我们检测了从1型糖尿病(T1DM)小鼠模型中分离的内皮细胞(ECs)的血管生成和代谢基因表达谱(GEP).
方法:从健康和链脲佐菌素(STZ)处理的小鼠的肾和心脏中分离ECs。然后提取RNA用于分子研究。通过微阵列检查了84个血管生成和84个AMP激活的蛋白激酶（AMPK）依赖性基因的GEP。实时PCR证实了在显著改变的基因中观察到的变化。通过免疫组织化学分析微血管密度(MVD),通过天狼星红组织学染色评估纤维化，并通过ELISA定量结缔组织生长因子（CTGF）。
结果：在T1DM动物的肾脏中ECs和MVD的相对百分比增加，而在糖尿病小鼠的心脏中观察到相反的趋势。因此，这些动物的大部分AMPK相关基因在肾脏中上调,在心脏中下调.血管生成性GEP显示Tgfβ存在显著差异，两个糖尿病器官中的Notch信号和Timp2。这些发现与血管生成组织学测定一致。与健康动物相比，糖尿病患者的两个器官的纤维化都增加了。
结论：总而言之，我们的研究结果表明,第一次,T1DM心脏和肾脏ECs呈现相反的代谢线索，伴随着不同的血管生成模式。这些发现使得能够开发针对糖尿病相关血管疾病的创新的器官特异性治疗策略。
方法:从健康和链脲佐菌素(STZ)处理的小鼠的肾和心脏中分离ECs。然后提取RNA用于分子研究。通过微阵列检查了84个血管生成和84个AMP激活的蛋白激酶（AMPK）依赖性基因的GEP。实时PCR证实了在显著改变的基因中观察到的变化。通过免疫组织化学分析微血管密度(MVD),通过天狼星红组织学染色评估纤维化，并通过ELISA定量结缔组织生长因子（CTGF）。
结果：在T1DM动物的肾脏中ECs和MVD的相对百分比增加，而在糖尿病小鼠的心脏中观察到相反的趋势。因此，这些动物的大部分AMPK相关基因在肾脏中上调,在心脏中下调.血管生成性GEP显示Tgfβ存在显著差异，两个糖尿病器官中的Notch信号和Timp2。这些发现与血管生成组织学测定一致。与健康动物相比，糖尿病患者的两个器官的纤维化都增加了。
结论：总而言之，我们的研究结果表明,第一次,T1DM心脏和肾脏ECs呈现相反的代谢线索，伴随着不同的血管生成模式。这些发现使得能够开发针对糖尿病相关血管疾病的创新的器官特异性治疗策略。
