Syndecan-1对大鼠心肌成纤维细胞生物学行为影响

Influence of syndecan-1 on biological behaviors of rat myocardial fibroblasts

目的评估syndecan-1(Sdc1)对大鼠心肌成纤维细胞生物学行为的影响,为心肌梗死(MI)后心室重构的防治提供依据。方法体外分离培养SD大鼠乳鼠心肌成纤维细胞,分别转染Sdc1 siRNA质粒和Sdc1高表达质粒,并设立空白对照组、无关序列和空载质粒作为阴性对照。采用流式细胞术检测细胞增殖,CCK-8法检测细胞活力,酶联免疫法(ELISA)检测胶原(羟脯氨酸)分泌能力及Transwell细胞迁移实验检测细胞迁移能力。结果Sdc1 siRNA组心肌成纤维细胞S期比例、增殖指数、细胞活力、胶原(羟脯氨酸)分泌量和迁移细胞数分别为(8.36±0.23)%、(13.2±0.5)%、(1.72±0.12)、(1162.3±60.4)pg/mL和170个/孔,明显高于空白对照和阴性对照组(P<0.01);Sdc1高表达质粒组心肌成纤维细胞S期比例、增殖指数、细胞活力、胶原(羟脯氨酸)分泌量和迁移细胞数分别为(6.48±0.22)%、(10.8±0.6)%、(1.30±0.11)、(346.8±52.1)pg/mL和50个/孔,明显低于空白对照和空载质粒组(P<0.01)。结论 Sdc1能够抑制心肌成纤维细胞的增殖、活力、迁移和分泌能力。

Objective To explore the influence of syndecan-1 （Sdcl） on biological behaviors of myocardial fibro- blasts,and to provide an experimental basis for the prevention of ventricular remodeling after myocardial infarction. Methods Myocardial fibroblasts of Sprague-Dawley （SD） rats were cultivated in vitro. Sdcl siRNA plasmid and Sdcl high expression plasmid were transferred into myocardial fibroblasts, respectively. Blank control, unconcerned sequence and empty vector plasmid were used as negative control. Flow cytometry was used to detect cell proliferation. Cholecysto- kinin octapeptide （CCK-8） was used to detect cell viability. Enzyme-linked immunosorbent assay （ELISA） was used to detect collagen （hydroxyproline） secretion. Transwell method was used to evaluate vertical migration of the cells. Results The proportion of S phase cells （8. 36% ±0. 23% ） ,proliferation index （ 13.2% ±0. 5% ） ,cell viability （ 1.72 ±0. 12）, collagen （hydroxyproline） secretion （ 1162. 3 ±60. 4 pg/ml）, and the number of migration cells （ 170 cells/hole） for the myocardial fibroblasts transferred with Sdcl siRNA plasmid were all higher than those in the myocardial fibroblasts of blank control and negative control group （all P 〈 0. 01 ）. The proportion of S phase cells （6.48% ±0. 22% ）, prolifera- tion index （ 10.8% ± 0.6% ）, cell viability （ 1.30 ± 0. 11 ）, collagen （ hydroxyproline ） secretion （ 346. 8 ± 52. 1 pg/ml）, and the number of migration cells （50 cells/hole） for the myocardial fibroblasts transferred with high Sdcl expression plasmid were all lower than those of the myocardial fibroblasts of blank control and empty vector plasmid group （P 〈 0. 01 for all）. Conclusion Sdcl can inhibit proliferation, activity, collagen secretion and migration capacity of myocardial fibroblasts.