摘要
目的 :观察钙化血管细胞凋亡的现象和细胞凋亡基因的表达 ,以认识细胞凋亡在血管钙化发病中的意义。方法 :利用维生素D3 (VitaminD3 )和尼古丁 (nicotine)制备大鼠血管钙化模型 ,检测血管钙含量、碱性磷酸酶(ALP)活性、45Ca摄入和血管骨钙素含量 ;应用原位缺口末端标记 (TUNEL)、免疫组织化学及DNALadder方法检测钙化血管的凋亡情况 ;应用免疫组织化学方法和3 H TdR参入方法观察钙化血管的增殖情况。结果 :血管的钙含量、ALP活性、45Ca摄取、骨钙素含量均显著增高。钙化血管中存在凋亡细胞 ,表现为细胞缩小、核浓缩、核碎裂等。TUNEL结果显示 ,钙化血管的凋亡指数比正常组高 36 % ,且Caspase 3和Bax的表达明显增强 ;Bcl 2的表达亦高于正常组 ;而NF κB基因表达比正常组低 2 6 %。钙化组可见清晰的DNA阶梯状条带 ,正常组则未出现。钙化组主动脉平滑肌细胞的增殖指数较正常组增加 ,3 H TdR参入量增加了 2 .5倍。结论
Objective:To observe the expression of the apoptosis genes in calcified vessels. Methods: Vascular calcification model was established in rats by using vitamin D 3 and nicotine. Vascular calcium content, alkaline phosphatase (ALP) activity, 45 Ca uptake and the content of osteocalcin in vessels were measured. Apoptosis was measured by TUNEL method, DNA ladder, and immunochemistry for the expression of apoptotic genes. Morphological changes were examined under microscope. Proliferation was measured by immunochemistry for PCNA and 3 H TdR incorporation. Results: Compared with those of control rats, the aortic calcium content,vascular ALP activity, 45 Ca uptake and the content of osteocalcin in rats of calcified group were obviously increased, TUNEL stainings were positive. The apoptosis index increased by 36%and the expression of Caspase 3 and Bax and Bcl 2 increased markedly, but the expression of NF κB was decreased. There was clear DNA ladder in calcified group, but not in the control group. The proliferation index was also increased in calcified group, and 3H TdR incorporation was increased by 2.5 times. Conclusion: The expression of apoptotic and proliferative genes were increased in calcified vessels.
出处
《北京大学学报（医学版）》
CAS
CSCD
北大核心
2003年第1期45-49,共5页
Journal of Peking University:Health Sciences
基金
国家重点基础研究发展规划项目 (G2 0 0 0 0 5 690 5 )基金资助~~
关键词
血管钙化
凋亡
增殖
细胞凋亡
Vascular calcification
Apoptosis
Proliferation
