本文選題：血管平滑肌細(xì)胞 + β-甘油磷酸�。� 參考：《臨床心血管病雜志》2017年01期

【摘要】：目的:探討骨形成蛋白-2(Bone morphogenetic protein 2,BMP-2)在高磷誘導(dǎo)的大鼠胸主動(dòng)脈血管環(huán)鈣化中的作用。方法:選取8~10周齡健康雄性SD大鼠,體外培養(yǎng)大鼠胸主動(dòng)脈血管環(huán),血管環(huán)按隨機(jī)數(shù)字表法隨機(jī)分為2組:正常對(duì)照組和高磷組。血管環(huán)培養(yǎng)7d和14d后,采用von Kossa染色及鄰甲酚酞絡(luò)合酮比色法檢測大鼠胸主動(dòng)脈血管環(huán)鈣化情況;免疫組織化學(xué)方法檢測血管環(huán)BMP-2的表達(dá)。體外采用組織塊貼壁法培養(yǎng)原代大鼠胸主動(dòng)脈平滑肌細(xì)胞,利用10mmol/Lβ-甘油磷酸制備血管平滑肌細(xì)胞鈣化模型,細(xì)胞隨機(jī)分為2組:正常對(duì)照組、高磷組(10mmol/Lβ-甘油磷酸)。采用茜素紅染色及鄰甲酚酞絡(luò)合酮比色法檢測細(xì)胞鈣化情況,RT-PCR和Western blot方法檢測細(xì)胞培養(yǎng)7d和14d的BMP-2mRNA及蛋白表達(dá),并觀察短時(shí)間內(nèi)不同時(shí)間點(diǎn)BMP-2蛋白表達(dá)情況。結(jié)果:體外血管環(huán)培養(yǎng)7d、14d后,與正常對(duì)照組相比,高磷組鈣含量明顯增加(P0.05);與7d高磷組血管環(huán)鈣含量相比,14d高磷組血管環(huán)鈣含量明顯增加(P0.05)。免疫組織化學(xué)結(jié)果顯示,與正常對(duì)照組相比,高磷組BMP-2表達(dá)增加(P0.05);與7d高磷組相比,14d高磷組血管環(huán)鈣含量明顯增加(P0.05)。細(xì)胞培養(yǎng)7d、14d后,與正常對(duì)照組相比,高磷組鈣鹽沉積及鈣含量明顯增高(P0.05);RTPCR和Western Blot顯示,與正常對(duì)照組相比,高磷組BMP-2mRNA及蛋白表達(dá)增加。進(jìn)一步動(dòng)態(tài)觀察BMP-2蛋白的表達(dá)變化,結(jié)果顯示正常對(duì)照組及高磷組BMP-2蛋白表達(dá)隨時(shí)間延長呈增強(qiáng)趨勢(P0.05)。結(jié)論:BMP-2可能參與了高磷誘導(dǎo)的血管鈣化的發(fā)生發(fā)展。
[Abstract]:Aim: to investigate the role of bone morphogenetic protein-2 bone morphogenetic protein 2 (BMP-2) in calcification of thoracic aortic rings in rats induced by high phosphorus. Methods: healthy male SD rats aged 8 ~ 10 weeks were selected and cultured in vitro. The rings were randomly divided into two groups: normal control group and high phosphorus group. After cultured for 7 and 14 days, the calcification of thoracic aortic rings in rats was detected by von Kossa staining and orthophenolphthalein complexone colorimetry, and the expression of BMP-2 in vascular rings was detected by immunohistochemistry. Primary rat aortic smooth muscle cells were cultured by tissue mass adherence in vitro. Vascular smooth muscle cells (VSMCs) calcified by 10mmol/L 尾 -glycerophosphoric acid (10mmol/L 尾 -glycerophosphate) were randomly divided into two groups: normal control group and high phosphorous group (10 mmol / L 尾 -glycerophosphate). The calcification of cells was detected by alizarin red staining and o-cresol phthalein complex ketone colorimetry. RT-PCR and Western blot methods were used to detect the expression of BMP-2mRNA and protein at 7 and 14 days of cell culture, and the expression of BMP-2 protein was observed at different time points in a short period of time. Results: compared with the normal control group, the calcium content in the high phosphorus group was significantly higher than that in the control group for 14 days, and the calcium content in the vascular ring in the high phosphorus group was significantly higher than that in the high phosphorus group on the 7th day after 14 days of culture, and the calcium content in the vascular ring in the high phosphorus group was significantly higher than that in the high phosphorus group on the 14th day. The results of immunohistochemistry showed that compared with the normal control group, the expression of BMP-2 increased in the high phosphorus group and the calcium content in the vascular ring of the high phosphorus group on the 14th day was significantly higher than that in the high phosphorus group on the 7th day. Compared with the normal control group, the calcium salt deposition and calcium content in the high phosphorus group were significantly higher than those in the control group. The expression of BMP-2mRNA and protein in the high phosphorus group was higher than that in the normal control group, and the expression of BMP-2mRNA and protein in the high phosphorus group was significantly higher than that in the control group. The expression of BMP-2 protein was observed dynamically. The results showed that the expression of BMP-2 protein in normal control group and high phosphorus group increased with time. Conclusion: BMP-2 may be involved in the development of vascular calcification induced by high phosphorus.
【作者單位】： 河北醫(yī)科大學(xué)第四醫(yī)院腎內(nèi)科;
【基金】：河北省自然科學(xué)基金資助項(xiàng)目(No:H2012206157) 河北省科技計(jì)劃項(xiàng)目(No:16397733D);河北省科技計(jì)劃項(xiàng)目(No:20150351);河北省科技計(jì)劃項(xiàng)目(No:20150310)
【分類號(hào)】：R692;R54

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