Development of organ specific transfection methods by using liposomes.

我们比较了由不同脂质组成的带正电脂质体的转染效率。在所使用的阳离子脂质体中，由TMAG、DOPE和DLPC组成的脂质体对质粒DNA的包裹效率要高得多，并且在没有或存在血清的情况下都能高效转染。阳离子脂质体的转染效率受脂质组成、脂质体类型或血清存在的影响。我们研究了阳离子脂质体是否能在爱情白血病病毒（BLV）长末端重复序列的控制下，有效地将白喉毒素a链（DT-A）的基因传递到BLV感染的细胞中，并且也适合在体内使用。阳离子脂质体可作为blv感染细胞的高效转染试剂，在体内可用于向blf感染细胞内递送DT-A基因。通过在大鼠角膜、虹膜、睫状体和视网膜内注射脂质体，可以实现功能基因高效、稳定的转移。我们发现，将特定种类脂质体携带的表达质粒载体滴入眼液中，可以将基因转移到大鼠的神经神经节细胞中，而不会引起任何炎症。这种非手术、方便的方式将基因传递到视网膜，将促进严重眼内疾病治疗的发展。用脂质体转染法研究了靶向原癌基因和细胞周期相关核蛋白的反义核苷酸对系膜细胞的抑制增殖作用。反义寡核苷酸抑制系膜细胞增殖。反义tgf - β和PDGF-B链抑制th1型肾炎模型细胞外基质的积累和系膜细胞的增殖。这些结果表明反义寡核苷酸作为一类新的治疗肾小球疾病的药物是可行的。

We compared the transfection efficiency of the positively charged liposomes composed of various lipids. In the cationic liposomes used, the liposomes composed of TMAG,DOPE,and DLPC showed an much higher efficiency for the plasmid DNA entrapment, and gave highly efficient transfection in the absence or presence of serum. The transfection efficiency of the cationic liposomes is affected by the lipid composition, type of liposomes, or the presence of serum.We investigated whether cationic liposomes are efficient at delivering the gene fo diphtheria toxin A-chain (DT-A) under the control of the long terminal repeats of lovine leukemia virus (BLV) in to BLV-infected cells and are also sutable for in vivo use. The cationic liposomes may be efficient transfection reagent for the BLV-infected cells and can be utilized for DT-A gene delivery into BLF-infected cells in vivo.Efficient and stable transfer of the functional gene could be achived by injection of liposomes in the cornea, iris, ciliary body, and retina of rats. We showed that instillation as eye drops of an expression plasmid vector carried by the specific kinds of liposomes could transfer the gene to the reinal ganglion cells of rat, without causing any inflammation. This non-surgical, convenient way for gene delivery to the reina would facilitate the development of treatment for vsrious inraocular desease.Anti-proliferative effect of antisense oliponucleotides targeting the proto-oncogenes and cell-cycle associated nuclear proteins on mesangial cells by tranfection of liposome methods were examined. The antisense oligonucleotides suppressed the mesagial cell proliferation. Antisense TGF-beta and PDGF-B chain inhibited accumulation of extracellular matrix and mesangial cell proliferation in Thy 1 glomelonephritis model. These results indicate the feasibility of antisense oligonucleotide as a novel class of therapeutic agents in the treatment of glomerular diseases.