高磷刺激大鼠血管平滑肌细胞后Cbfα1及Sox9基因动态变化的研究  

The dynamic change of Cbfα1 and Sox9 of rat vascular smooth muscle cells induced by β-glycerophosphate

作　　者：白亚玲[1] 徐金升[1] 石博[1] 张俊霞[1] 崔立文[1] 张慧然[1] 张胜雷[1] 

机构地区：[1]河北医科大学第四医院肾内科,石家庄050011

出　　处：《临床心血管病杂志》2014年第6期542-545,共4页Journal of Clinical Cardiology

基　　金：河北省自然科学基金(No:H2012206157)

摘　　要：目的:观察高磷刺激大鼠血管平滑肌细胞(VSMCs)后表型转化相关基因的动态变化,探讨VSMCs的表型转化方向。方法:组织块贴壁法原代培养大鼠胸主动脉VSMCs,经形态学和免疫细胞化学鉴定后,将其随机分为空白对照组和β-甘油磷酸组,RT-PCR法同管扩增检测VSMCs核心结合因子α1(Cbfα1)、性别决定区Ybox 9(Sox9)、Ⅱ型胶原a1(ColⅡa1)及Ⅹ型胶原a1(ColⅩa1)mRNA在不同时间点的动态表达情况。结果:随着高磷刺激时间的延长,Cbfα1mRNA的表达呈先上升,后下降,之后再次升高的趋势(P<0.05),即呈"峰-谷-峰"样;而Sox9则表现为"单峰"(P<0.05),且其达峰时间处于Cbfα1的谷值和第2峰值之间。作为Sox9下游基因的ColⅡa1的变化与之类似,而ColⅩa1则在初期维持基础的低水平,而在Cbfα1第2次出现峰值后表达显著升高(P<0.05)。结论:高磷刺激后Cbfα1和Sox9及其下游Ⅱ型和Ⅹ型胶原mRNA的动态变化符合软骨细胞分化的特点,进而推测VSMCs类似于成骨/软骨前体细胞、增生软骨细胞、肥厚软骨细胞的分化过程可能参与了β-甘油磷酸诱导的表型转化。Objective：To explore the effect of β-glycerophosphate on the transdifferentiation associated gene in rat vascular smooth muscle cells （VSMCs）. Method：Primary VSMCs, obtained from rat thoracic aorta and identi- fied with morphology and immunocytochemistry, were randomly divided into control and β-glycerophosphate group. RT-PCR was performed to assess the variation trend of core binding factor α1 （Chfα1）, sex-determining region Y-box 9 （Sox9）, type Ⅱ collagen al （Col Ⅱ al） and type Ⅹ collagenal （Col Ⅹ al） at different time points. Result：After stimulation of 10 mmol/L β-glycerophosphate, the variation trend of Cbfα1 characteristic with peak- valley-peak, the expression of Cbfα1 increased gradually at first, then fell down, and increased again （P〈0.05）. However, Sox9 showed a ＂single peak＂ （P〈0.05）, and the peak of Sox9 was located between the valley and the second peak of Cbfα1. Consistent with the variation trend of Sox9, Col Ⅱ al also showed a ＂single peak＂ （P〈 0.05）. While Col Ⅹ al was low expressed at the beginning and then significantly increased after the second peak of Cbfα1 （P〈0.05）. Conclusion：After stimulation of β-glycerophosphate, the dynamic change of Cbfα1, Sox9 and their downstream gene Col Ⅱ and Col Ⅹ were coincidence with their expression characteristic in the differeciation of ehondrocyte. The transdifferentiation of VSMCs induced by β-glycerophosphate was similar with differentiation of ostochondroprecursors, proliferating chondrocytes and hypertrophic chondrocytes.

关 键 词：β-甘油磷酸 血管平滑肌细胞 表型转化 

分 类 号：R543.5[医药卫生—心血管疾病]