骨髓间充质干细胞及Klotho基因移植对慢性缺氧大鼠心脏重构的影响
作者:
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(1. 昆明总医院地方干部病房,昆明 650032;2. 云南省干细胞工程实验室,昆明 650032)

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R541;R363

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国家科技支撑计划项目(2014BI01B0);国家973计划项目(2012CB5181060);云南省科技计划重点项目(2013CA005)


Effects of bone marrow mesenchymal stem cells and Klotho gene transplantation on cardiac remodeling in chronic hypoxic rats
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(1. Department of Civilian Cadre’s Ward, Kunming General Hospital of Kunming Military Command, Kunming 650032, China;2. Yunnan Provincial Laboratory of Stem Cell Engineering, Kunming 650032, China)

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    摘要:

    目的 观察骨髓间充质干细胞(BMSCs)及抗衰老基因(Klotho基因)移植对慢性缺氧大鼠心脏重构的影响。方法 建立慢性缺氧28 d大鼠心脏重构模型,分为模型组、BMSCs组和Klotho组,并另设正常对照组不给予缺氧处理。分离、培养、鉴定大鼠BMSCs;重组慢病毒介导Klotho基因。模型建立结束后分别通过尾静脉移植BMSCs及Klotho基因至慢性缺氧大鼠体内,4周后用ELISA法检测血清和心肌中Ⅰ型前胶原羟基端肽(PⅠCP)和Ⅲ型前胶原羟基端肽(PⅢCP)的浓度,HE染色法观察心肌细胞的形态,MASSON染色检测心肌胶原含量,Tunel检测心肌细胞凋亡率。采用SPSS 17.0软件进行数据处理。计量资料以均数±标准差([AKx-D]±s)表示,组间比较采用t检验。结果 移植BMSCs和Klotho基因4周后,BMSCs组和Klotho组血清和心肌中的PⅠCP及PⅢCP浓度、胶原容积分数(CVF)和心肌细胞凋亡率均较模型组显著降低(P<0.01)。与Klotho组比较,BMSCs组心肌细胞凋亡率显著降低(P<0.01)。结论 BMSCs和Klotho基因移植均能有效逆转心脏重构,BMSCs在抗心肌细胞凋亡方面优于Klotho基因。

    Abstract:

    Objective To investigate the effect of transplantation of bone marrow mesenchymal stem cells (BMSCs) and anti-aging gene (Klotho gene) on cardiac remodeling in rats under chronic hypoxic condition. Methods Rat model of cardiac remodeling was established in a hypoxic chamber of 10% oxygen for 28 d. The rats were divided into model group, BMSCs group and Klotho group, and another normal control group for isolation, culture and identification of BMSCs. Recombinant lentival vector of Klotho gene was constructed. After the establishment of the model, the BMSCs and lentival vector of Klotho were transplanted into the chronic hypoxia rats by tail vein. In 4 weeks after the infusion, the contents of procollagenⅠcarboxy-terminal propeptide (PⅠCP) and PⅢCP in the serum and myocardial tissues were detected by ELISA. The morphology of cardiomyocytes was observed by HE staining, the content of myocardial collagen was detected by MASSON staining, and the apoptotic rate of myocardial cells was detected by Tunel. SPSS statistics 17.0 was used to analyze the data. The measurement data were expressed as mean±standard deviation ([AKx-D]±s), and Student’s t test was employed for the comparison between groups. Results After 4 weeks’ transplantation of BMSCs and Klotho gene, the serum and myocardial contents of PⅠCP and PⅢCP, collagen volume fraction (CVF) and apoptotic rate of cardiomyocytes were significantly lower in the BMSCs group and Klotho group than the model group (P<0.01). Compared with the Klotho group, the BMSCs group had significantly lower apoptotic rate of myocardial cells (P<0.01). Conclusion BMSCs and Klotho gene transplantation can both effectively reverse cardiac remodeling, and BMSCs is superior to Klotho gene in resisting myocardial cell apoptosis.

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李慧萍,曹帅,王恩阳,朱向情,尹娜,魏玲,潘兴华.骨髓间充质干细胞及Klotho基因移植对慢性缺氧大鼠心脏重构的影响[J].中华老年多器官疾病杂志,2017,16(8):609~614

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  • 收稿日期:2017-05-15
  • 最后修改日期:2017-06-26
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  • 在线发布日期: 2017-08-25
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