摘要
目的观察沙格列汀对高糖环境下成骨细胞增殖、分化和凋亡的影响,为糖尿病性骨质疏松患者选择合理的降糖药物提供理论依据。方法取成骨细胞MC3T3-E1置于a-MEM培养基中常规培养、传代。取传3代的对数生长期细胞,随机分为正常对照组、高糖组、高糖联合沙格列汀组,即刻更换a-MEM培养基,正常对照组更换为葡萄糖浓度5.5 mmol/L的培养液,高糖组更换为葡萄糖浓度30.0 mmol/L的培养液,高糖联合沙格列汀组更换为葡萄糖浓度30.0 mmol/L+沙格列汀1μmol/L的培养液,继续培养48 h。收集各组细胞,分别采用CCK-8法和流式细胞术检测细胞增殖能力和细胞凋亡率,ELISA法检测碱性磷酸酶(ALP)活性,qRT-PCR法检测成骨细胞分化相关基因Ⅰ型胶原(COL-Ⅰ)、成骨特异性转录因子2(Runx2)、骨钙蛋白(OCN)、骨桥蛋白(OPN)mRNA表达。结果与正常对照组比较,高糖组、高糖联合沙格列汀组细胞增殖能力、ALP活性及成骨细胞分化相关基因COL-Ⅰ、Runx2、OCN、OPN mRNA相对表达量均明显降低,细胞凋亡率均明显升高(P均<0.05);与高糖组比较,高糖联合沙格列汀组细胞增殖能力、ALP活性和成骨细胞分化相关基因COL-Ⅰ、Runx2、OCN、OPN mRNA相对表达量均明显升高,细胞凋亡率均明显降低(P均<0.05)。结论沙格列汀能在一定程度上逆转高糖环境对小鼠成骨细胞增殖、分化和凋亡的影响,为糖尿病性骨质疏松患者的降糖治疗提供了理论依据。
Objective To investigate the effects of saxagliptin on the proliferation,differentiation,and apoptosis of osteoblasts in high glucose environment,so as to provide the theoretical basis for the selection of rational hypoglycemic agents for patients with diabetic osteoporosis. Methods Osteoblasts MC3T3-E1 were cultured and passaged in a-MEM medium. The cells of 3 generations in the logarithmic phase were randomly divided into the normal control group,high glucose group,and high glucose ＋ saxagliptin group. The a-MEM medium was immediately replaced with 5. 5 mmol/L glucose medium in the normal control group,the a-MEM medium was immediately replaced with 30. 0 mmol/L glucose medium in the high glucose group,and the a-MEM medium was immediately replaced with 30. 0 mmol/L glucose ＋ 1 μmol/L saxagliptin medium in the high glucose ＋ saxagliptin group. MC3T3-E1 osteoblasts were then cultured in different culture mediums for48 h. CCK-8 method and flow cytometry were used to detect the proliferation ability and apoptosis rate,ELISA assay was used to detect alkaline phosphatase（ ALP） activity,and qRT-PCR was used to detect the mRNA expression of COL-Ⅰ,Runx2,OCN and OPN. Results Compared with the normal control group,the proliferation,ALP activity and expression of COL-ⅠmRNA,Runx2 mRNA,OCN mRNA,and OPN mRNA were significantly decreased,and the apoptosis rate was significantly increased in the high glucose group and high glucose ＋ saxagliptin group（ all P〈0. 05）. Compared with the high glucose group,the proliferation,ALP activity and expression of COL-Ⅰ mRNA,Runx2 mRNA,OCN mRNA,and OPN mRNA were significantly increased,and the apoptosis rate was significantly decreased in the high glucose ＋ saxagliptin group（ all P〈0. 05）. Conclusion Saxagliptin can reverse the effects of high glucose environment on proliferation,differentiation,and apoptosis of MC3T3-E1 osteoblasts to a certain extent,which can be used for hypoglycemic therapy for patients with diabetic osteoporosis.
出处
《山东医药》
CAS
北大核心
2017年第28期16-19,共4页
Shandong Medical Journal
基金
国家自然科学基金资助项目(81400788)
