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-  2015 

Triton X-100对脂质体介导的BMP-2基因转染大鼠BMSCs的作用

DOI: doi:10.7507/1002-1892.20150016

Keywords: BMSCs, Triton X-100, 脂质体, 基因治疗

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Abstract:

目的探讨Triton X-100对脂质体介导的BMP-2基因转染大鼠BMSCs的作用。 方法8周龄 Wistar大鼠1只,雄性,体质量120 g。取Wistar大鼠股骨、胫骨骨髓,采用贴壁培养法分离培养BMSCs并鉴定。取第3代BMSCs,采用MTT法筛选Triton X-100适宜浓度。实验分成3组,实验组:BMP-2+脂质体+0.010% Triton X-100+BMSCs,常规转染组:BMP-2+脂质体+BMSCs,空白对照组:BMSCs+血清培养基。转染后48 h,倒置荧光显微镜观察各组绿色荧光蛋白表达情况;转染后72h,采用实时荧光定量PCR检测BMP-2 mRNA表达。 结果0.010%Triton X-100既能使BMSCs保持一定的活力,又可达到一定的作用效果,故确定其为适宜浓度。转染后48 h,倒置荧光显微镜下见实验组和常规转染组均有绿色荧光蛋白表达,空白对照组无绿色荧光蛋白表达。转染后72 h,实验组BMP-2 mRNA相对表达量为5.94±0.12,常规转染组为4.99±0.08,差异有统计学意义(t=360.28,P=0.02)。实验组转染效率较常规转染组平均提高了19%。 结论0.010%Triton X-100可以促进脂质体介导的BMP-2基因转染大鼠BMSCs,并提高其转染效率

References

[1]  1. 催少千, 范广宇, 李建军, 等. 腺病毒介导的BMP-2基因转染兔骨髓基质干细胞对其增殖、成骨分化的影响. 东南大学学报:医学版, 2009, 28(1):22-25.
[2]  10. 张昌明, 吴永寿, 余宏. 阳离子脂质体基因载体的研究进展. 医学综述, 2009, 15(12):1768-1770.
[3]  11. Shin J, Quails MM, Boomer JA, et al. An eficient new route to plasmenyl-typo lipids:synthesis an d cytotoxicity of a plasmenylcholine an alogue of the an titumor ether lipid ET-18-OMe. J Am Chem Soc, 2001, 123(3):508-509.
[4]  15. Managit C, Kawakami S, NishikawaM, et al. Targeted and sustmned drug delivery using PEGylated galactosylated liposomes. Int J Pharm, 2003, 266(1-2):77-84.
[5]  16. Mohammadabadi MR, El-Tamimy M, Gianello R, et al. Supramolecular assemblies of zwitterionic nanoliposomepolynucleotide complexes as gene transfer vectors:Nanolipoplex formulation and in vitro characterisation. J Liposome Res, 2009, 19(2):105-115.
[6]  20. 沈明, 蒋海鹰. TritonX-100在培养细胞免疫组化染色中的应用. 临床与实验病理学杂志, 1998, 14(2):197-199.
[7]  22. Johnsson M, Bergstrand N. Phase behavior of DOPE/TritonX100(reduced) in dilute aqueous solution:aggregate structure and pHdependence. Colloids Surf B Biointerfaces, 2004, 34(2):69-76.
[8]  3. 赵冰冰, 张玮, 王琪, 等. TIZ基因全长扩增和携带其基因的真核表达载体的构建. 广西医科大学学报, 2012, 29(1):45-47.
[9]  4. Bryksin AV, Matsumura I. Overlap extension PCR cloning:a simple and reliable way to create recombinant plasmids. Biotechniques, 2010, 48(6):463-465.
[10]  5. Minmer M, Simanek E. Nonviral Vectors for Gene Delivery. J Chem Rev, 2009, 109(2):259-302.
[11]  6. 魏云波, 高平, 刘书花, 等. 阳离子脂质体转染效率的主要影响因素. 北方药学, 2012, 9(11):86.
[12]  7. Hu YL, Huang B, Zhang TY, et al. Mesenchymal stem cells as a novel carrier for targeted delivery of gene in cancer therapy based on nonviral transfection. Mol Pharm, 2012, 9(9):2698-2709.
[13]  8. 陈彦祥, 乔卫红, 刘栋良, 等. 阳离子脂质体的转染机制及转染效率影响因素. 生物工程学报, 2007, 23(5):776-780.
[14]  12. Guosen H, Min F, Xin L, et al. Design, synthesis and in vitro evaluation of a novel "stealth" polymeric gene vector. Int J Pharm, 2008, 350(1):344-350.
[15]  17. 王冰, 张树彪, 周集体, 等. 阳离子脂质体介导的基因转移机制. 中国组织工程研究与临床康复, 2011, 15(8):1459-1462.
[16]  18. Ko IK, Ziady A, Lu S, et al. Acid-degradable cationic methacrylamide polymerized in the presence of plasmid DNA as tunable non-viral gene carrier. Biomaterials, 2008, 29(28):3872-3881.
[17]  19. Amidzadeh Z, Behbahani AB, Erfani N, et al. Assessment of different permeabilization methods of minimizing damage to the adherent cells for detection of intracellular RNA by flow cytometry. Avicenna J Med Biotechnol, 2014, 6(1):38-46.
[18]  2. 夏德林, 刘道华, 贾娟, 等. 兔骨髓基质干细胞体外分离培养及生物学特性研究. 中国美容医学, 2011, 20(8):1251-1254.
[19]  9. Zhang Y, Rong Qi X, Gao Y, et al. Mechanisms of co-modified liver-targeting liposomes as gene delivery carriers based on cellular uptake and antigens inhibition effect. J Control Release, 2007, 117(2):281-290.
[20]  13. Incani V, Tunis E, Clements BA, et al. Palmitic acid substitution on cationic polymers for effective delivery of plasmid DNA to bone marrow stromal cells. J Biomed Mater Res, 2007, 81(2):493-504.
[21]  14. Zabner J, Fasbender AJ, Moninger T, et al. Cellular and molecular barrier to gene transfer by a cationic lipid. J Biol Chem, 1995, 270(32):18997-19007.
[22]  21. 姜笃银, 钱春华, 周兵, 等. 不同脱细胞方法对无细胞真皮基质抗原成分的影响. 中国临床康复, 2005, 9(6):35-37.

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