- 學位論文
口服聚合微膠體投予脊髓神經保護基因
In vivp oral neuroprotective genes delivery into mouse spinal cord by nano block biopolymeric micelles
摘要
脊髓損傷後的照顧和修復一直是近年來急欲研究的課題。雖然過去的方法皆以藥物 Methyprednisolone (MP) 來控制和治療，但是人們卻發現這樣的方法會造成較大之副作用。然而在過去的研究中顯示，脊髓損傷後,去髓鞘化與寡突狀細胞的死亡和軸索的傷害有很大的關係。因此，本實驗主要目的針對BCL-xL ，一與抑制寡突狀細胞壞死之相關性基因，和利用一段可以專一性表現在寡突狀細胞的啟動子MBP (myelin basic protein)做為標的基因，且利用polymeric Micelle (PM) 進行基因傳遞的方式架構出一個質體DNA (pMBP-Bcl-xL-EGFP)來進行基因治療評估。同時也觀察在給予 MP 的治療劑量下，同時觀察 Bcl-xL 之抗細胞壞死的基因表現量。在初步結果發現中，直接從脊髓組織之螢光的表現 EGFP 或是從Bcl-xL 蛋白質西方點墨法的或是由mRNA Bcl-xL 定量Real-time PCR ，質體DNA是可以在口服六個dose下，48小時後有其表現。而在給予 MP 的實驗中，我們也發現到，有 PM 包覆的 MP 和在 mRNA 的Bcl-xL 與，似乎有延長作用時間的效果。
關鍵字
聚合微膠體並列摘要
Abstract The treatment and repair of the spinal cord injury have been crucial subjects in recent years. In the past, MP was used to be treated with the acute spinal cord injury (SCI), though with great side effects.Recently research shows that after the spinal cord injury are very great relations of the demyelination with the death of oligodendrocyte cell and the damage of axon. To obviate the multiple genetic mechanisms of MP, the anti-apoptotic protein Bcl-xL gene could be one approvals of developing novel gene delivery systems to avoid the advance side of effect of MP. We have a nano-size of PM with Bcl-xL gene (pMBP-Bcl-xL-EGFP), driven by oligodendrcytes-specific promoters of Myelin basic protein(MBP) into the spinal cord. Our finding indicate those two days after six doses of oral delivery of 40 μg with 3 times a day, gene expression was observed at 48 h in the spinal cord by immunohistological observation, Bcl-xL western-blotting, m RNA of Bcl-xL by real time PCR, Furthermore, using PM incorporated with MP, we did observe the mRNA of Bcl-xL have sustained in the spinal cord.