成骨细胞条件性FoxO1基因敲除糖尿病小鼠模型的建立
Establishment of diabetic mouse model with conditional knockout of FoxO1 in osteoblasts
作者：熊毅, 宫苹, 伍颖颖
Author：Xiong Yi, Gong Ping, Wu Yingying
收稿日期：2017-09-15 年卷（期）页码：2018,45(2):170-176
期刊名称：国际口腔医学杂志
Journal Name：International Journal of Stomatology
关键字：叉头转录因子,条件性基因敲除,糖尿病,
Key words：forkhead box protein O,conditional knock-out,diabetes mellitus,
基金项目：国家自然科学基金（81400543,81571008）; 四川大学优秀青年学者科研基金（A类）（2017SCU04A21）
中文摘要
目的 建立成骨细胞条件性FoxO1基因敲除的糖尿病小鼠模型,并进行初步的表型研究。方法 设计基因敲除策略,获得野生型和纯合小鼠。提取鼠尾组织DNA,经聚合酶链式反应（PCR）扩增后鉴定其基因型。分别提取小鼠原代成骨细胞mRNA和蛋白质,以及其他组织的mRNA,利用实时荧光定量聚合酶链式反应（RT-PCR）和Western blot技术检验FoxO1的表达。在不同时间点监测小鼠体重、血糖,并进行胰岛素耐受实验。结果 成功建立FoxO1基因敲除小鼠模型并诱发糖尿病,糖尿病小鼠从第2周开始体重明显低于正常小鼠。在糖尿病条件下,纯合小鼠血糖值明显低于野生型小鼠,其胰岛素敏感性明显高于野生型小鼠。结论 成功建立了成骨细胞条件性FoxO1基因敲除小鼠模型;FoxO1基因敲除可改善糖尿病小鼠的高糖血症,可能是由于FoxO1基因敲除增加了糖尿病小鼠的胰岛素敏感性。
英文摘要
ObjectiveThis study aims to establish a diabetic mouse model with conditional knockout ofFoxO1 in osteoblasts and preliminary explore the phenotypes.MethodsWild type (WT) and gene knockout mice were obtained according to the designed reproductive strategy. DNA was extracted from mouse tail and amplified by real-time polymerase chain reaction (RT-PCR). The mice phenotypes were also detected. The mRNA and protein levels ofFoxO1 in osteoblasts and other tissues were obtained and detected by RT-PCR and Western blot analysis. Mice weight and fasting blood glucose were measured. Insulin tolerance test (ITT) was conducted to assess the role ofFoxO1 in glucose metabolism.ResultsDiabetic mouse models with conditional knockout ofFoxO1 in osteoblasts were successfully established. The body weight of diabetic mice was significantly lower than that of normal mice at 2 weeks. The fasting blood glucose in diabetic gene knockout mice was lower than that in diabetic WT mice. In addition, ITT showed thatFoxO1 knockout promoted insulin sensitivity in diabetic gene knockout mice compared with that in diabetic WT mice.ConclusionDiabetic mouse model with conditional knockout ofFoxO1 in osteoblasts was successfully established.FoxO1 knockout ameliorated hyperglycaemia, which may be accounted for the increased insulin sensitivity in diabetic gene knockout mice.
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