Klotho基因转染骨髓间充质干细胞移植对慢性心力衰竭大鼠心肌纤维化的影响

发布时间：2018-02-11 08:10

本文关键词： Klotho基因骨髓间充质干细胞心力衰竭心肌纤维化　出处：《中国动脉硬化杂志》2017年06期 　论文类型：期刊论文

【摘要】：目的观察Klotho基因(抗衰老基因)修饰的骨髓间充质干细胞(BMSC)移植对慢性心力衰竭大鼠心肌纤维化的影响。方法分离、培养、扩增大鼠骨髓间充质干细胞。慢病毒介导Klotho转染至BMSC中,逆转录-聚合酶链反应(RT-PCR)检测BMSC中Klotho mRNA的表达。制备大鼠慢性心力衰竭模型,随机分为正常对照组、模型对照组、增强型绿色荧光蛋白-BMSC组(EGFP-BMSC组)、EGFP-Klotho-BMSC组。细胞移植28天后多普勒超声检测心功能,荧光显微镜观察细胞存活及分布,HE染色观察心肌组织病理变化。Masson染色检测心肌胶原沉积含量和心肌纤维化程度。结果 Klotho转染BMSC后,BMSC中Klotho mRNA表达明显增多(P0.05)。移植28天后超声心动图结果显示,与EGFP-BMSC组及模型对照组比较,EGFP-Klotho-BMSC组左心室射血分数(LVEF)提高(P0.05)。EGFP-BMSC组及EGFP-Klotho-BMSC组心肌纤维化程度较模型对照组减轻(P0.05),EGFP-KlothoBMSC组较EGFP-BMSC组心肌纤维化改善更为明显(P0.05)。结论 Klotho基因联合BMSC治疗可进一步抑制心肌纤维化,减少心肌间质胶原沉积,改善心功能。
[Abstract]:Objective to observe the effect of transplantation of bone marrow mesenchymal stem cells modified with Klotho gene on myocardial fibrosis in rats with chronic heart failure. Rat bone marrow mesenchymal stem cells were amplified. Lentivirus-mediated Klotho was transfected into BMSC, and the expression of Klotho mRNA in BMSC was detected by reverse transcription-polymerase chain reaction (RT-PCR). The rat model of chronic heart failure was randomly divided into normal control group and model control group. EGFP-Klotho-BMSC and EGFP-Klotho-BMSC were detected by Doppler ultrasound 28 days after transplantation. The survival and distribution of cells were observed by fluorescence microscope. The pathological changes of myocardial tissue were observed by HE staining. Masson staining was used to detect myocardial collagen deposition and myocardial fibrosis. Results after Klotho transfection into BMSC, the expression of Klotho mRNA increased significantly. After 28 days, echocardiography showed that, Compared with the EGFP-BMSC group and the model control group, the left ventricular ejection fraction (LVEF) of EGFP-Klotho-BMSC group increased P0.05N. EGFP-BMSC group and EGFP-Klotho-BMSC group decreased the degree of myocardial fibrosis compared with the model control group. Conclusion Klotho gene combined with BMSC can improve myocardial fibrosis more obviously than that of EGFP-BMSC group. Treatment can further inhibit myocardial fibrosis, Reduce myocardial interstitial collagen deposition and improve cardiac function.
【作者单位】：成都军区昆明总医院地方干部病房;云南省干细胞工程试验室;
【基金】：973计划项目(2012CB518100)
【分类号】：R542.23

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