过氧化物酶体增殖物激活受体γ抑制高磷诱导血管平滑肌细胞钙化  

PPAR γ inhibits calcification in vascular smooth muscle cells induced by high phosphate

作　　者：刘亮[1] 肖堂利[1] 余彦霖[1] 何婷[1] 徐新丽[1] 管旭[1] 杨可[1] 张道海[1] 郑昌玲[1] 赵景宏[1] 

机构地区：[1]第三军医大学新桥医院肾内科,全军肾脏病中心,重庆400037

出　　处：《第三军医大学学报》2016年第2期124-128,共5页Journal of Third Military Medical University

基　　金：国家自然科学基金面上项目(81270290)~~

摘　　要：目的探讨过氧化物酶体增殖物激活受体γ(peroxisome proliferators activated receptorγ,PPARγ)对高磷诱导的小鼠血管平滑肌细胞(vascular smooth muscle cells,VSMCs)钙化的抑制作用。方法体外培养小鼠血管平滑肌细胞,先分正常对照组、高磷组(HP,2.6 mmol/L),观察高磷对VSMCs的影响,再分为正常对照组、高磷组(HP,2.6 mmol/L)、罗格列酮组(RGL,10μmol/L)、高磷(2.6 mmol/L)+罗格列酮(10μmol/L)组(HP+RGL),观察PPARγ激动剂罗格列酮对高磷诱导的VSMCs钙化的抑制作用。茜素红S(alizarin red S)染色观察高磷对细胞钙盐沉积的影响。Western blot检测PPARγ、血管平滑肌22alpha蛋白标志物(SM22α)、骨形成蛋白2(BMP2)和成骨特异性转录因子2(Runx2)的表达变化。结果与正常对照组相比,高磷处理后的VSMCs的钙盐沉积明显升高[(0.08±0.02)vs(0.19±0.03),P<0.01],成骨细胞标志物BMP2[(0.26±0.02)vs(0.74±0.03),P<0.01]及Runx2表达[(0.29±0.03)vs(0.91±0.04),P<0.01)],明显升高。同时PPARγ[(0.93±0.04)vs(0.58±0.02),P<0.05]及平滑肌细胞标志物SM22α表达减少[(1.02±0.09)vs(0.77±0.03),P<0.05],而加入罗格列酮后,高磷状态下VSMCs的钙盐沉积明显降低[(0.19±0.02)vs(0.12±0.03),P<0.05],PPARγ[(0.63±0.04)vs(0.85±0.03),P<0.05]和SM22α[(0.69±0.02)vs(0.99±0.03),P<0.01]表达明显上升,相反,BMP2[(1.02±0.04)vs(0.48±0.05),P<0.01]及Runx2[(1.00±0.06)vs(0.67±0.03),P<0.01]的表达则明显受抑。结论高磷诱导VSMCs向成骨细胞分化和钙化可能与下调PPARγ的表达有关,而罗格列酮可通过激活PPARγ抑制高磷状态下VSMCs钙化的发生。Objective To determine the role of peroxisome proliferators activated receptor γ（ PPARγ） in calcification of vascular smooth muscle cells（ VSMCs） induced by high phosphate. Methods Cultured mouse VSMCs were divided into normal phosphate（ NP） group and high phosphate（ HP,2. 6 mmol / L）group,and the effect of HP on VSMCs was observed. Then the VSMCs were divided into NP group,HP（ 2. 6 mmol/L） group,rosiglitazone（ RGL,10 μmmol/L） group,and HP ＋ RGL（ 2.6 mmol/L ＋10 μmmol/L）group,and the inhibitory effect of PPARγ agonist RGL on the calcification of VSMCs induced with HP was observed. Alizarin red S staining was used to observe HP-induced calcification of VSMCs. Expression level of PPARγ,SM22α,BMP2,and Runx2 were measured by Western blotting. Results Compared with the NP group,calcium salt deposition of HP-treated VSMCs was higher（ 0. 08 ± 0. 02 vs 0. 19 ± 0. 03,P〈0. 01）,the expression levels of BMP2（ 0. 26 ± 0. 02 vs 0. 74 ± 0. 03,P〈0. 01） and Runx2（ 0. 29 ± 0. 03 vs 0. 91 ±0. 04,P〈0. 01） were increased significantly,and the expression levels of PPARγ（ 0. 93 ± 0. 04 vs 0. 58 ±0. 02,P〈0. 05） and SM22α（ 1. 02 ± 0. 09 vs 0. 77 ± 0. 03,P〈0. 05） were decreased significantly. After incubation with RGL,calcium salt deposition of HP-treated VSMCs was reduced（ 0. 19 ± 0. 02 vs 0. 12 ±0. 03,P〈0. 05）,the expression levels of PPARγ（ 0. 63 ± 0. 04 vs 0. 85 ± 0. 03,P〈0. 05） and SM22α（ 0. 69 ± 0. 02 vs 0. 99 ± 0. 03,P〈0. 01） were markedly elevated,and the expression levels of BMP,（ 1. 02 ± 0. 04 vs 0. 48 ± 0. 05,P〈0. 01） and Runx2（ 1. 00 ± 0. 06 vs 0. 67 ± 0. 03,P〈0. 01） were dropped markedly. Conclusion HP-induced osteoblast differentiation and calcification of VSMCs may be related with the down-regulation of PPARγ,and HP-induced calcification of VSMCs can be inhibited by RGL.

关 键 词：平滑肌细胞 PPARγ 血管钙化 高磷血症 

分 类 号：R322.12[医药卫生—人体解剖和组织胚胎学]