摘要
目的探讨炎症对高磷所致大鼠血管平滑肌细胞钙化的影响及可能机制。方法原代培养大鼠血管平滑肌细胞,分为空白对照组、高磷组、炎症组及高磷+炎症组。高磷组、高磷+炎症组在加入4.8 mmol/L磷的基础培养基中培养,建立高磷诱导的大鼠血管平滑肌细胞钙化模型。高磷+炎症组加入10μg/m L细菌脂多糖(LPS),炎症组只加入10μg/m L细菌脂多糖(LPS),空白对照组在基础培养基中培养。各组均培养48 h后,检测培养液C反应蛋白(CRP)水平、细胞碱性磷酸酶(ALP)活性及钙沉积量;Realtime-PCR法检测细胞骨形成蛋白-2(BMP-2)、肿瘤抑制基因4(Smad4)、肌肉片段同源盒2(Msx2)及成骨细胞特异性转录因子(Osterix)mRNA相对表达量。结果高磷组、炎症组及高磷+炎症组培养液中CRP水平、细胞ALP活性及钙沉积量均高于空白对照组(P均<0.05),且高磷+炎症组培养液中CRP水平、细胞ALP活性及钙沉积量均高于高磷组及炎症组(P均<0.05),炎症组培养液中CRP水平和细胞钙沉积量均低于高磷组(P均<0.05)。高磷组、炎症组及高磷+炎症组细胞中BMP-2、Smad4、Msx2及Osterix mRNA相对表达量均高于空白对照组(P均<0.05),且高磷+炎症组细胞中BMP-2、Smad4、Msx2及Osterix mRNA相对表达量均高于高磷组及炎症组(P均<0.05),炎症组细胞中Smad4、Msx2及Osterix mRNA相对表达量均低于高磷组(P均<0.05)。结论炎症可显著加重高磷所致的大鼠血管平滑肌细胞钙化,其作用机制可能与上调BMP-2、Smad4、Msx2、Osterix表达及ALP活性等有关。
Objective To study the effects and possible mechanisms of inflammation on the calcification of rat vascu-lar smooth muscle cells （VSMCs）primarily caused by high phosphate.Methods Rat VSMCs were primarily cultured and then were divided into the control （Ctr）group,high phosphate （Pi）group,inflammation （LPS）group,high phosphate and inflammation （Pi＋LPS）group.Rat VSMCs in the Pi group and Pi＋LPS group were cultured in basal medium contai-ning 4.8 mmol/L Pi to establish the calcification models successfully.Rat VSMCs in the Pi ＋LPS group were cultured in basal medium containing 4.8 mmol/L Pi and 10 μg/mL LPS.Rat VSMCs in the LPS group were cultured in basal medium only containing 10 μg/mL LPS.Rat VSMCs in the Ctr group were cultured in basal medium.The C-reactive protein （CRP）level of culture medium,alkaline phosphatase （ALP）activity and calcium deposition of cells in each group were respectively measured after 48-hour culture;real-time PCR was used to detect the relative mRNA expression of bone mor-phogenetic protein-2 （BMP-2）,Smad4,Msx2 and Osterix.Results The CRP level of culture medium,ALP activity and calcium deposition of cells in the Pi group,LPS group and Pi＋LPS group were higher than those of the Ctr group （all P〈0.05）.The CRP level of culture medium,ALP activity and calcium deposition of cells in the Pi＋LPS group were higher than those in the Pi group and LPS group （all P〈0.05）.The CRP level of culture medium and calcium deposition of cells in the LPS group were lower than those of the Pi group （all P〈0.05）.The relative mRNA expression of BMP-2,smad4,＆amp;nbsp;msx2,and Osterix in the Pi group,LPS group and Pi＋LPS group was higher than that of the Ctr group （all P〈0.05 ）. The relative mRNA expression of BMP-2,smad4,msx2 and Osterix in the Pi＋LPS group was higher than that of the Pi group amd LPS group （all P〈0.05 ）.The relative mRNA expression of smad4,msx2 and Osterix in the LPS group was lower than that of the Pi group （all P〈0.05）.Conclusions The calcification of rat VSMCs cultured in vitro could be in-duced by both high phosphate and inflammation.Inflammation combined with high phosphate could aggravate the calcifica-tion of rat VSMCs cultured in vitro significantly.The mechanisms may be ralted with the up-regulation of BMP-2,Smad4, Msx2,Osterix expression and the ALP activity.
出处
《山东医药》
CAS
北大核心
2016年第46期15-19,共5页
Shandong Medical Journal
基金
四川省教育厅项目基金资助项目(09ZC118)
关键词
炎症
高磷
平滑肌细胞
血管钙化
终末期肾病
inflammation
high phosphate
smooth muscle cells
vascular calcification
end-stage renal disease
