本文選題：格列齊特　切入點(diǎn)：MC3T3-E1細(xì)胞　出處：《鄭州大學(xué)》2015年碩士論文　論文類型：學(xué)位論文

【摘要】：目的探討格列齊特在正糖環(huán)境中對(duì)小鼠成骨細(xì)胞樣細(xì)胞MC3T3-E1增殖、分化及凋亡的影響方法體外培養(yǎng)小鼠成骨細(xì)胞樣細(xì)胞MC3T3-E1,分為正常對(duì)照組和不同濃度格列齊特組(10μmol/L、20μmol/L、100μmol/L),格列齊特干預(yù)48h后,采用CCK-8檢測(cè)細(xì)胞增殖率,流式細(xì)胞儀檢測(cè)細(xì)胞凋亡率,熒光定量PCR檢測(cè)成骨細(xì)胞分化標(biāo)志物I型膠原(COL-I)、骨橋蛋白(OPN)和成骨因子Runt相關(guān)基因2(Runx2)基因的表達(dá),Western blots檢測(cè)凋亡蛋白(Bax)和抗凋亡蛋白(Bcl-2)的表達(dá)。結(jié)果(1)與對(duì)照組相比,不同濃度格列齊特均能促進(jìn)成骨細(xì)胞增殖,20μmol/L時(shí)促進(jìn)增殖的作用最強(qiáng),10μmol/L格列齊特組、20μmol/L格列齊特組、100μmol/L格列齊特組與對(duì)照組相比細(xì)胞增殖率依次增加16.6%,20.2%,13.7%(P0.05)。(2)對(duì)照組、10μmol/L格列齊特組、20μmol/L格列齊特組、100μmol/L格列齊特組的早期凋亡率分別為:8.83%,5.47%,2.87%,7.33%。與對(duì)照組相比,不同濃度格列齊特均降低成骨細(xì)胞早期凋亡率(P0.05),20μmol/L抑制凋亡作用最顯著。(3)與對(duì)照組相比,不同濃度格列齊特(10μmol/L、20μmol/L、100μmol/L)均能促進(jìn)功能蛋白COL-1、OPN、Runx2的m RNA的表達(dá)(P0.05),20μmol/L格列齊特作用最顯著,且COL-1、OPN與Runx2基因的表達(dá)呈顯著正相關(guān)(相關(guān)系數(shù)r分別為0.899、0.854,均P0.01)。(4)對(duì)照組、10μmol/L格列齊特組、20μmol/L格列齊特組、100μmol/L格列齊特組Bcl-2/β-actin的值依次為1.22±0.025,1.48±0.035,1.81±0.066,1.30±0.015;Bax/β-actin的值依次為0.83±0.046,0.62±0.015 0.53±0.0360.72±0.025。與對(duì)照組相比,不同濃度格列齊特組Bcl-2蛋白表達(dá)量增高,Bax蛋白表達(dá)量降低(P0.05),20μmol/L作用最顯著。Bcl-2/Bax比值與細(xì)胞早期凋亡率呈顯著負(fù)相關(guān)(相關(guān)系數(shù)r=-0.935,P0.01)。結(jié)論格列齊特能促進(jìn)正糖環(huán)境下培養(yǎng)的成骨細(xì)胞樣細(xì)胞MC3T3-E1的增殖、分化,抑制其凋亡,對(duì)成骨細(xì)胞起保護(hù)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 鹵0.0251.48 鹵0.036 鹵1.81 鹵0.066 鹵1.30 鹵0.015Bax/ 尾 -actin, 0.83 鹵0.0460.62 鹵0.015 0.53 鹵0.0360.72 鹵0.02525 鹵0.0251.48 鹵0.035 鹵0.065 鹵0.015 Bax/ 尾 -actin in the control group, and the expression of Bcl-2/ 尾 -actin in the control group was 0.83 鹵0.046 0.62 鹵0.015 0.53 鹵0.0360.72 鹵0.025251.48 鹵0.035 鹵0.036 鹵1.30 鹵0.015 鹵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.
【學(xué)位授予單位】：鄭州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：R580;R587.1

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