摘要
目的:观察血管紧张素Ⅱ(AngⅡ)对培养人血管平滑肌细胞钙化的影响,以探讨血管钙化发生的机制。方法:体外培养人动脉平滑肌细胞,用β甘油磷酸(βGP)诱导细胞钙化。培养细胞分4组:正常对照组;βGP组:βGP终浓度为10mmol·L-1;AngⅡ组:AngⅡ终浓度为10-7mol·L-1;βGP+AngⅡ组:同时加入相同浓度的βGP和AngⅡ,连续培养10d。采用Vonkossa染色、细胞层钙沉积定量和细胞内碱性磷酸酶(ALP)活性测定判断钙化程度,用流式细胞仪测细胞周期分布和细胞凋亡率。结果:βGP组细胞Vonkossa染色见大量黑色颗粒状沉积物弥漫分布于细胞层,βGP+AngⅡ组细胞之间散在分布黑色沉着点;βGP+AngⅡ组细胞钙沉积显著低于βGP组[分别为(1.336±0.096)和(2.056±0.441)μmol·L-1,P<0.01];βGP+AngⅡ组细胞内ALP活性显著低于βGP组[分别为(277.57±87.81)和(691.27±128.06)IU·L-1,P<0.01]。正常对照组与AngⅡ组细胞比较,Vonkossa染色、细胞钙沉积量和ALP活性均无显著性差异。流式细胞仪结果显示,βGP+AngⅡ组细胞凋亡率显著低于βGP组,G0/G1期细胞比例降低,S期的比例升高(P<0.05)。结论:10-7mol·L-1的AngⅡ可抑制钙化血管平滑肌细胞的体外钙化进程。其作用可能与抑制ALP活性和促进细胞增殖、抑制凋亡有关。
Objective: To evaluate the effect of Angiotension-Ⅱ(AngⅡ) on calcium deposition and alkaline phosphatase(ALP) activity in the human vascular smooth muscle cells(HVSMCs), and to investigate the mechanism of AngⅡ to inhibit calcification of HVSMCs. Methods: The cell culture of human vascular smooth muscle was performed. The β-glycerophospahte sodium salt was used to induce cultured HVSMCs to calcify to establish a vascular calcification model in vitro. Cells were divided into four groups: the control, β-GP, AngⅡ and β-GP+ AngⅡ group. 10 -7 mol·L -1 AngⅡ was added into the culture media. Calcification was confirmed by the Vokossa staining. The calcium deposition of the cellular layer and the ALP activity of intracellular were quantified during a 10 days culturing process. The cell cycle and apoptosis rate was analyzed. Results: After 10 days of culture, the calcium deposition in β-GP+ AngⅡ group was less than in β-GP group[(1.336±0.096) μmol·L -1 versus (2.056± 0.441) μmol·L -1, P<0.01]. The cellular ALP activity in β-GP+ AngⅡ group was less than in β-GP group[(691.27±128.06) IU·L -1 versus (277.57±87.81) IU·L -1, P<0.01]. This process was time-dependent. The flow cytometeric DNA analysis revealed that 10 -7 mol·L -1 AngⅡ caused significantly decreasing of apoptosis rate, descending of G 0/G 1 phase and increasing of S phase compared with calcified HVSMCs. Conclusion: The angiotension-Ⅱ decreases calcium deposition of calcified HVSMCs. This may be through decreasing ALP activity, stimulating proliferation and inhibiting apoptosis of HVSMCs.
出处
《武汉大学学报（医学版）》
CAS
2005年第3期284-288,i005,共6页
Medical Journal of Wuhan University
基金
湖北省教育厅科研基金资助(编号:2003A003)
关键词
血管紧张素Ⅱ
钙化
血管平滑肌细胞
AngiotensionⅡ
Calcification
Vascular Smooth Muscle Cells
