不同浓度厄贝沙坦对前成骨细胞分化和矿化的影响

Effects of different concentrations of irbesartan on the differentiation and mineralization of preosteoblasts

背景:有研究发现,血管紧张素Ⅱ受体拮抗剂对骨的保护作用强于较血管紧张素转换酶抑制剂。目的:观察不同浓度血管紧张素Ⅱ受体拮抗剂厄贝沙坦对小鼠前成骨细胞分化和矿化的影响。方法:选取对数生长期小鼠前体成骨细胞株MC3T3-E1,分4组培养,分别加入含0(对照),0.001,0.01,0.1 mmol/L厄贝沙坦的成骨诱导培养基培养。诱导培养10 d,采用碱性磷酸酶染色观察细胞分化情况;诱导培养21 d,采用茜素红染色观察细胞矿化情况;诱导培养1,4,7,14,21 d,RT-PCR检测细胞成骨细胞内骨钙素、碱性磷酸酶、Runt相关转录因子2mR NA的相对表达量。结果与结论:(1)碱性磷酸酶染色:0.001,0.01,0.1 mmol/L厄贝沙坦组碱性磷酸酶活性均高于对照组(P<0.05),其中以0.01 mmol/L效果最明显;(2)茜素红染色:0.001,0.01,0.1 mmol/L厄贝沙坦组钙结节数量及面积均高于对照组(P<0.05),其中以0.01 mmol/L效果最明显;(3)RT-PCR检测:0.01 mmol/L厄贝沙坦组诱导培养不同时间点的碱性磷酸酶、Runt相关转录因子2、骨钙素mR NA相对表达量均高于对照组(P<0.05);(4)结果表明:0.01 mmol/L厄贝沙坦可显著促进成骨细胞的分化和矿化。

BACKGROUND： Angiotensin II receptor antagonists have been found to exerct a stronger protective effect on bone than angiotensin converting enzyme inhibitors. OBJECTIVE： To investigate the effect of different concentrations of irbesartan（angiotensin II receptor antagonist）on the differentiation and mineralization of mouse preosteoblasts. METHODS： Mouse preosteoblast cell lines MC3T3-E1 in logarithmic phase were selected and cultured in the osteogenic induction medium containing 0（control group）, 0.001, 0.01, 0.1 mmol/L irbesartan, respectively. Ten days later, the cell differentiation was observed by alkaline phosphatase staining. The mineralization was observed by alizarin red staining after 21 days of culture. m RNA expressions of osteocalcin, alkalinephosphatase and Runt-associated transcription factor 2 in osteoblasts were detected by real-time PCR at 1, 4, 7, 14 and 21 days of culture. RESULTS AND CONCLUSION： The activity of alkaline phosphatase in all the irbesartan groups（0, 0.001, 0.01, 0.1） was higher than that in the control group（P〈0.05）, which was the most obvious in 0.01 mmol/L. The number and area of calcium nodules in each irbesartan group were significantly higher than those in the control group（P〈0.05）, especially in 0.01 mmol/L. Compared with the control group, 0.01 mmol/L irbesartan significantly upregulated the m RNA expressions of osteocalcin, alkaline phosphatase and Runt-associated transcription factor 2（P〈0.05）. These results suggest that 0.01 mmol/L irbesartan significantly promotes the differentiation and mineralization of osteoblasts.