Zhang Ye, Bradshaw-Pierce Erica L, Delille Alexandra, Gustafson Daniel L, Anchordoquy Thomas J
Department of Pharmaceutical Sciences, School of Pharmacy, University of Colorado Health Sciences Center, Denver, CO 80262, USA.
J Pharm Sci. 2008 Jan;97(1):237-50. doi: 10.1002/jps.21076.
To evaluate the in vivo biodistribution and expression of DOTAP-Chol/DNA complexes (lipoplexes) with different in vitro serum stability, quantitative real-time PCR, in vitro luciferase expression and whole body luminescence imaging were used. In general, less tissue biodistribution, lower luciferase expression and whole body luminescence were observed for DOTAP:Chol (mol/mol 1:4)/DNA lipoplexes which had higher in vitro serum stability as compared to DOTAP:Chol (mol/mol 1:1)/DNA lipoplexes. Plasmid DNA biodistribution and expression were mainly confined to the lungs, and the results suggest that in vitro serum stability may serve as a predictor of transfection in the lung. No correlation between plasmid DNA tissue biodistribution and gene expression was observed by simultaneous determination of the level of plasmid DNA tissue biodistribution and gene expression. While high doses of the formulation possessing increased in vitro serum stability did exhibit reduced entrapment in the lung, no corresponding increase in the plasmid levels of other tissues was observed. However, this formulation did show increased accumulation in tumors that was not further enhanced by PEGylation.
为评估具有不同体外血清稳定性的DOTAP-胆固醇/DNA复合物（脂质体）的体内生物分布和表达情况，采用了定量实时PCR、体外荧光素酶表达及全身发光成像技术。总体而言，与DOTAP:胆固醇（摩尔/摩尔1:1）/DNA脂质体相比，具有较高体外血清稳定性的DOTAP:胆固醇（摩尔/摩尔1:4）/DNA脂质体在组织中的生物分布较少、荧光素酶表达较低且全身发光较弱。质粒DNA的生物分布和表达主要局限于肺部，结果表明体外血清稳定性可作为肺部转染的预测指标。通过同时测定质粒DNA组织生物分布水平和基因表达水平，未观察到质粒DNA组织生物分布与基因表达之间存在相关性。虽然具有增强体外血清稳定性的高剂量制剂在肺部的截留确实减少，但未观察到其他组织中质粒水平相应增加。然而，该制剂在肿瘤中的蓄积增加，聚乙二醇化并未进一步增强这种作用。
