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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 investigate the inhibitory effect of peroxisome proliferators activated receptor γ (PPARγ) on the calcification of vascular smooth muscle cells (VSMCs) induced by high phosphorus in mice. Methods Vascular smooth muscle cells (VSMCs) were cultured in vitro and divided into normal control group and high phosphorus group (HP, 2.6 mmol / L) / L), rosiglitazone group (RGL, 10μmol / L) and high phosphorus (2.6mmol / L) + rosiglitazone group (HP + RGL) Inhibition of high phosphorus-induced calcification of VSMCs. Alizarin red S staining was used to observe the effects of high phosphorus on the deposition of cellular calcium. Western blot was used to detect the expression of PPARγ, 22alpha protein marker (SM22α), bone morphogenetic protein 2 (BMP2) and osteoblast-specific transcription factor 2 (Runx2). Results Compared with the normal control group, calcium deposition in VSMCs treated with high phosphorus significantly increased ([(0.08 ± 0.02) vs (0.19 ± 0.03), P <0.01] and osteoblast marker BMP2 [(0.26 ± 0.02 ) vs (0.74 ± 0.03), P <0.01] and Runx2 expression [(0.29 ± 0.03) vs (0.91 ± 0.04), P <0.01)]. Meanwhile, the expression of SM22α in PPARγ [(0.93 ± 0.04) vs (0.58 ± 0.02), P <0.05] and the smooth muscle cell marker decreased [(1.02 ± 0.09) vs (0.77 ± 0.03), P <0.05] (P <0.05), PPARγ [(0.63 ± 0.04) vs (0.85 ± 0.03), P <0.05], and the calcium deposition of VSMCs under high phosphorus stress was significantly lower than that of the control (P <0.01), and the expression of Runx2 [(1.00 ± 0.06) and SM22α [(0.69 ± 0.02) vs ) vs (0.67 ± 0.03), P <0.01] was significantly inhibited. Conclusion High-phosphorus-induced differentiation of VSMCs into osteoblasts and calcification may be related to the down-regulation of PPARγ expression. Rosiglitazone can inhibit the calcification of VSMCs under high-phosphorus stress by activating PPARγ.