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目的研究钙化发生过程中细胞DNA损伤响应分子表达水平的变化。方法半定量方法测定钙化细胞中p21、p53、c-abl的mRNA水平,通过免疫印迹法检测p53和chk1磷酸化水平。结果血管平滑肌细胞在10 mmolβ-磷酸甘油酯的诱导下,随着时间的延长,在时间点24,,6 d时,p21、p53、c-abl的mRNA水平上升,第8天后mRNA水平下降。在前6 d中,p53和chk1磷酸化呈逐步上升趋势。结论在钙化过程的前期,p21、p53、c-abl的mRNA水平上升,p53和chk1磷酸化水平也呈上升趋势,这表明钙化的发生和DNA损伤响应分子有关。
Objective To study the changes of DNA damage response molecule expression during calcification. Methods The mRNA levels of p21, p53 and c-abl in calcified cells were determined by semi-quantitative method. The phosphorylation levels of p53 and chk1 were detected by immunoblotting. Results The mRNA levels of p21, p53 and c-abl mRNA in vascular smooth muscle cells increased with time after induced by 10 mmol β-glyceryl trinitrate at 24, 6 and 6 days, and decreased after 8 days. In the first 6 days, p53 and chk1 phosphorylation showed a gradual upward trend. Conclusion In the early stage of calcification, the mRNA levels of p21, p53 and c-abl increased, and the phosphorylation of p53 and chk1 also showed an upward trend, indicating that the occurrence of calcification was related to the DNA damage response molecule.