格列齐特对小鼠成骨细胞样细胞MC3T3-E1增殖、分化及凋亡的影响

发布时间：2018-03-13 10:03

本文选题：格列齐特　切入点：MC3T3-E1细胞　出处：《郑州大学》2015年硕士论文　论文类型：学位论文

【摘要】：目的探讨格列齐特在正糖环境中对小鼠成骨细胞样细胞MC3T3-E1增殖、分化及凋亡的影响方法体外培养小鼠成骨细胞样细胞MC3T3-E1,分为正常对照组和不同浓度格列齐特组(10μmol/L、20μmol/L、100μmol/L),格列齐特干预48h后,采用CCK-8检测细胞增殖率,流式细胞仪检测细胞凋亡率,荧光定量PCR检测成骨细胞分化标志物I型胶原(COL-I)、骨桥蛋白(OPN)和成骨因子Runt相关基因2(Runx2)基因的表达,Western blots检测凋亡蛋白(Bax)和抗凋亡蛋白(Bcl-2)的表达。结果(1)与对照组相比,不同浓度格列齐特均能促进成骨细胞增殖,20μmol/L时促进增殖的作用最强,10μmol/L格列齐特组、20μmol/L格列齐特组、100μmol/L格列齐特组与对照组相比细胞增殖率依次增加16.6%,20.2%,13.7%(P0.05)。(2)对照组、10μmol/L格列齐特组、20μmol/L格列齐特组、100μmol/L格列齐特组的早期凋亡率分别为:8.83%,5.47%,2.87%,7.33%。与对照组相比,不同浓度格列齐特均降低成骨细胞早期凋亡率(P0.05),20μmol/L抑制凋亡作用最显著。(3)与对照组相比,不同浓度格列齐特(10μmol/L、20μmol/L、100μmol/L)均能促进功能蛋白COL-1、OPN、Runx2的m RNA的表达(P0.05),20μmol/L格列齐特作用最显著,且COL-1、OPN与Runx2基因的表达呈显著正相关(相关系数r分别为0.899、0.854,均P0.01)。(4)对照组、10μmol/L格列齐特组、20μmol/L格列齐特组、100μmol/L格列齐特组Bcl-2/β-actin的值依次为1.22±0.025,1.48±0.035,1.81±0.066,1.30±0.015;Bax/β-actin的值依次为0.83±0.046,0.62±0.015 0.53±0.0360.72±0.025。与对照组相比,不同浓度格列齐特组Bcl-2蛋白表达量增高,Bax蛋白表达量降低(P0.05),20μmol/L作用最显著。Bcl-2/Bax比值与细胞早期凋亡率呈显著负相关(相关系数r=-0.935,P0.01)。结论格列齐特能促进正糖环境下培养的成骨细胞样细胞MC3T3-E1的增殖、分化,抑制其凋亡,对成骨细胞起保护作用。
[Abstract]:Objective to investigate the effect of gliclazide on MC3T3-E1 proliferation of mouse osteoblast like cells in normal glucose environment. Methods Murine osteoblast like cells MC3T3-E1 were cultured in vitro and divided into two groups: normal control group and different concentration gliclazide group (10 渭 mol / L ~ (20) 渭 mol / L ~ (10) 渭 mol 路L ~ (-1) 路L ~ (-1)). After 48 h of intervention by gliclazide, cell proliferation rate was measured by CCK-8 and apoptosis rate was measured by flow cytometry. Fluorescence quantitative PCR was used to detect the expression of osteopontin (OPN) and osteogenic factor Runt related gene (2G Runx2). Western blots was used to detect the expression of apoptotic protein (Bax) and anti-apoptotic protein (Bcl-2). Different concentrations of gliclazide could promote the proliferation of osteoblasts at 20 渭 mol/L. The effect of 10 渭 mol/L gliclazide group on proliferation was the strongest. The cell proliferation rate of the 10 渭 mol/L gliclazide group and the control group was increased by 16. 6% 20. 2 and 13. 07% in comparison with the control group. The cell proliferation rate of the 10 渭 mol/L gliclazide group increased in turn compared with the control group (P 0. 05. 0. 05. 2) the cell proliferation rate of the 10 渭 mol/L gliclazide group increased in turn compared with that of the control group. The early apoptotic rate of 20 渭 mol/L gliclazide group and 100 渭 mol/L gliclazide group was: 8.83 and 5.475.472.87and 7.333.Compared with the control group, Compared with the control group, different concentrations of gliclazide (10 渭 mol / L ~ 20 渭 mol / L ~ (100 渭 mol / L)) could promote the expression of m RNA of the functional protein COL-1OOPN- Runx2 and the expression of m RNA of P0.05- (20 渭 mol / L) gliclazide was the most significant, compared with the control group, at different concentrations of gliclazide (P _ (0.05)) 20 渭 mol / L ~ (-1), the expression of m RNA of the osteoblast was significantly increased by different concentrations of gliclazide (P _ (0.05)), compared with that of the control group. There was a significant positive correlation between the expression of COL-1OOPN and Runx2 gene (r = 0.899n0.854, all P 0.01g / L). The values of Bcl-2/ 尾 -actin in 10 渭 mol/L gliclazide group were 1.22 卤0.0251.48 卤0.036 卤1.81 卤0.066 卤1.30 卤0.015Bax/ 尾 -actin, 0.83 卤0.0460.62 卤0.015 0.53 卤0.0360.72 卤0.02525 卤0.0251.48 卤0.035 卤0.065 卤0.015 Bax/ 尾 -actin in the control group, and the expression of Bcl-2/ 尾 -actin in the control group was 0.83 卤0.046 0.62 卤0.015 0.53 卤0.0360.72 卤0.025251.48 卤0.035 卤0.036 卤1.30 卤0.015 卤0.015 Bax/ 尾 -actin, respectively. In different concentration of gliclazide group, the expression of Bcl-2 protein increased and the expression of Bax protein decreased significantly. Bcl-2 / Bax ratio was negatively correlated with the early apoptosis rate of cells. Conclusion Gliclazide can promote the culture in normal glucose environment, and the ratio of Bcl-2 / Bax is negatively correlated with the rate of early cell apoptosis (r = 0. 935, P 0. 01, r = 0. 935, P 0. 01, P 0. 01). The proliferation of osteoblast like cell MC3T3-E1, Differentiation, inhibition of apoptosis and protection of osteoblasts.
【学位授予单位】：郑州大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R580;R587.1

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