心力衰竭大鼠骨髓间充质干细胞形态及功能的改变

Morphological and functional changes of mesenchymal stem cells of rats with heart failure

目的研究心力衰竭大鼠的骨髓间充质干细胞(BMSC)在细胞形态、蛋白表达及细胞因子分泌能力的变化。方法本文采用部分缩窄腹主动脉法制备大鼠压力超负荷性慢性心力衰竭模型,分离和培养模型组和假手术组大鼠的BMSC,待细胞传代到第6代,观察细胞的形态和蛋白表达的变化情况。分别以1×108/L的细胞密度培养模型组和假手术组大鼠BMSC,72 h后收集上清。采用酶联免疫吸附试验(ELISA)技术检测BMSC培养上清中的HGF、IGF-1、PDGF、SCF、FGF和VEGF的含量。组间比较采用t检验,多组间比较采用单因素方差分析(ANOVA)。结果模型组BMSC的最小细胞直径、单个细胞的平均面积及单个细胞的蛋白表达量均显著低于假手术组,差异具有统计学意义(P<0.01)。模型组细胞培养上清中的SCF和PDGF的浓度分别为(6.06±1.18)ng/L和(1.25±0.32)mg/L,明显低于假手术组(12.56±1.15)ng/L和(3.52±0.23)mg/L,而模型组的VEGF浓度为(652.44±83.19)ng/L,明显高于假手术组(419.97±67.85)ng/L,差异具有统计学意义(P<0.01)。结论心力衰竭大鼠的BMSC与健康大鼠相比,细胞形态发生显著改变,单个细胞的蛋白表达量及细胞因子SCF、PDGF的分泌能力均降低,但VEGF的分泌量上升。

Objective To study the variation on bone marrow mesenchymal stem cells(BMSC)of rats with heart failure in morphology, protein expression and the production of cytokines. Methods The rat model of ventricular hypertrophy and heart failure was established by partially banding abdominal aorta. BMSC of sham-operated group and model group were isolated and cultured in vitro. BMSC was observed and the protein expression was detected by immunocytochemistry. The supernatants of BMSC(1×108/L)were collected after culture for 72 h and HGF, IGF-1,PDGF, SCF, FGF and VEGF were measured by enzymelinked immunosorbent assay(ELISA). Results TThe minimum diameter, the expression of CD29 and CD44, and average area of single BMSC in the heart failure group were obviously decreased than those in the sham-operated group(P < 0.01).The levels of SCF and PDGF in heart fialure group were(6.06 ± 1.18)ng/L and(1.25 ± 0.32)mg/L respectively, obviously lower than those in the sham-operated group(12.56 ± 1.15)ng/L and(3.52 ± 0.23)mg/L.However,VEGF(652.44 ± 83.19)ng/L was obviously higher than that in sham-operated group[(419.97 ± 67.85)ng/L, P < 0.01]. Conclusions The morphology of BMSC after heart failure was significantly changed, and production of SCF and PDGF was decreased compared with normal rats. But the secretion of VEGF was increased compared with the control.