體內口服基因聚合微膠體之評估

本研究之初步目標，在於評估以PEO-PBLA嵌段式聚合微膠體作為體內口服含有一段LacZ基因的質體DNA之載體。以每天投與3個劑量的方式，在口服投與6個劑量，48小時之後能達到最高的基因表現。而且，基因表現可以在十二指腸的絨毛隱窩和杯狀細胞以及胃組織的隱窩細胞經由X-gal定性染色的方法觀察到。此外，在全血和肝臟也可測得基因表現。另一方面，若預先給予EDTA後，甚至於在腦及睪丸亦可測得基因表現。另外，利用體外穿透試驗，評估此嵌段式共聚合物與質體DNA所形成的微膠體製劑在不同濃度的質體DNA下對十二指腸的影響。其結果顯示，在低濃度下(<130μg/μl)所測得的擬穿透係數，並未隨濃度的增加而增加，且達到(5.75±0.37) X 10-5 cm/s。此外，在分別以RGD和EDTA處置的結果得知，其擬穿透係數各為(0.95±0.57) X 10-5 cm/s以及(29.8±5.7) X 10-5 cm/s，可以推測此嵌段式共聚合物與質體DNA所形成的微膠體製劑可能是經由吞噬作用(endocytosis)的途徑進入組織，而且可以經開啟緊密結合區(tight junction)的作用來增加質體DNA進入組織的量，進而增加其基因轉染效率。總而言之，本實驗室以PEO-PBLA嵌段式聚合微膠體作為體內口服的基因載體，可以達到安全以及有效的基因表現

關鍵字

基因傳遞；非病毒載體；口服；聚合微膠體

並列摘要

The primary objective of this study was to investigate the feasibility of using PEO-PLBA non-ionic copolymeric micelles as a carrier for oral gene delivery of plasmid DNA with the LacZ gene in vivo. After six doses of oral delivery 3 times a day, the most intense gene expression was observed at 48 h. Reporter gene expression was detected around the villi, crypts, and goblet cells of duodenum and crypt cells of stomach tissues. Also reporter gene was expressed in the circulating blood, stomach and liver organs. Furthermore, brain and testis organs were observed the marker geneexpression after EDTA treatment. Assessment of the effect of the copolymers with plasmid on the duodenum in the in vitro penetration study showed that the P was dose independent at low concentrations (< 130 ?g/?l) and reach plateau at (5.75 ? 0.37) x 10-5 cm/s. Furthermore, after RGD and EDTA treatment, the transport mechanisms of the plasmid/block copolymeric micelles were found to occur through endocytosis in tissues by enhancement through tight junction pathways with P values to (0.95 ?0.57), and(29.8 5.7) x 10-5 cm/s, respectively. Taken together, we reported an efficient and stable gene transfer with PEO-PPO-PEO polymeric micelles through oral delivery can be achieved in mice.

並列關鍵字

gene delivery ； non-viral vector ； oral ； polymeric micelles

參考文獻

1. Anderson, W. F., Blaese, R. M., and Culver, K. (1990). The ADA human gene therapy clinical protocol: Points to consider response with clinical protocol, July 6, 1990. Hum Gene Ther. 1, 331-362.

2. Anderson, W. F. (1998). Human gene therapy. Nature 392, 25-30.

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5. Ronald G. C. (1995). Transfer of genes to humans: early lessons and obstacles to success. Science 270, 404-410.

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體內口服基因聚合微膠體之評估

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