基质谷氨酸蛋白在家猪主动脉瓣间质细胞钙化中的表达  

Matrix Gla protein expression in the process of calcification in aortic valve interstitial cells of susscrota domestica

作　　者：金洋[1] 杨军政[1] 付庆林[1] 韩培立[1] 

机构地区：[1]新乡医学院第一附属医院心血管外科,河南卫辉453100

出　　处：《新乡医学院学报》2016年第3期182-185,189,共5页Journal of Xinxiang Medical University

摘　　要：目的研究家猪主动脉瓣间质细胞（AVIC）体外钙化诱导过程中基质谷氨酸蛋白（MGP）的变化。方法处死健康成年雄性家猪6只,取下心脏,分离、培养AVIC,并进行细胞表型鉴定。取2~5代AVIC,分为在标准培养基中培养的对照组与在钙化诱导培养基（标准培养基内加入钙化诱导剂）中培养的钙化诱导组,分别采用茜素红染色及Western blot、半定量聚合酶链反应技术检测2组细胞在1、7、14 d的钙化情况及MGP在蛋白及mRNA水平的表达情况。结果对照组细胞在7、14 d出现少量钙化结节[每孔（2.38±0.97）个、（9.47±1.59）个],钙化诱导组细胞在7、14 d出现明显钙化结节[每孔（37.55±6.70）个、（78.29±9.14）个];钙化诱导组细胞在7、14 d钙化情况明显高于对照组（P〈0.05）。在7、14 d,钙化诱导组细胞内活化型MGP蛋白水平及MGP mRNA表达水平明显低于对照组（P〈0.05）。结论在钙化诱导条件下,AVIC内钙化抑制剂MGP抑制钙化能力下调。Objective To inveatigate the changes of matrix Gla protein（ MGP） expression in the process of calcification in aortic valve interstitial cell（ AVIC） of susscrota demestica. Methods Six healthy male susscrota domestica were sacrificed and AVIC were isolated from the aortic valve leaflets of them,then the cells were cultured,and the type of the isolated cells was identified before further research. AVIC grew in culture until they reached passages 2 ～ 5 before experimentation.Then,the cells were divided into control group and calcification group according to the absence or presence of osteogenic additives. After 1 day,7,14 days,alizarin red staining and assay were used to stain and measure the calcium deposit,and Western blot and semi-quantity polymerase chain reaction were used to detected the levels of MGP protein and MGP mRNA in AVIC in the two groups. Results After 7 days and 14 days,calcified nodules were formed in both control group[（ 2. 38 ± 0. 97） per hole,（ 9. 47 ± 1. 59） per hole] and calcification group[（ 37. 55 ± 6. 70） per hole,（ 78. 29 ± 9. 14） per hole],quantification of calcified nodules was obviously higher in calcification group than that in control group（ P 〈0. 05）. The levels of activated MGP protein and MGP mRNA were significantly lower in calcification group compared with that in control group after 7 days and 14 days（ P 〈0. 05）. Conclusion The anti-calcification effects of MGP is deficient in the process of calcification.

关 键 词：基质谷氨酸蛋白 主动脉瓣 瓣膜间质细胞 瓣膜钙化 家猪 

分 类 号：R654.2[医药卫生—外科学]