評估類黃酮以固體脂質微粒製劑為運輸載體的生物活性

Investigation the bioactivity of solid lipid nanoparticles as carriers of flavonoids

蔡雅婷(Ya-Ting Tsai)

指導教授 ： 蔡義弘

高雄醫學大學/藥學院/藥學研究所/碩士(2009年)

摘要

類黃酮是天然的抗氧化劑，本實驗挑選了五類共八種類黃酮(apigenin、baicalein、epicatechin、genistein、kaempferol、myricetin、naringenin、quercetin)，篩選了三種皆可同時包埋八種類黃酮的處方(formulation-1、formulation和formulation-3)，製成固體脂質微粒製劑，分別加入五種不同癌細胞(HepG2、Hep3B、Ca9-22、MRC-5、MCF7、MDA-MB-231)中，測試其體外癌細胞之毒殺能力。毒殺結果發現八種類黃酮包覆於處方F-3中之抗癌效果明顯優於其他兩種(F-1、F-2)以及未製成劑型的solution，處方F-3獲得有效的抗癌能力，且IC50值遠低於其餘兩處方，可見添加之賦形劑對劑型活性有很大影響。
為進一步探討類黃酮製成固體脂質微粒製劑的生物活性，因此再選用處方F-3與未製成劑型的solution進行抗氧化與清除自由基的活性實驗測試，但實驗結果處方F-3之活性遠低於solution，並不如預期有效，推測可能是因處方F-3之包埋率皆高達90%以上，藥物完全被包覆於處方中無法發揮作用，且此實驗設計只偵測加入藥物後80秒的反應，而類黃酮無法於如此短的時間中將藥物釋出，因此以此試驗無法看出類黃酮固體脂質微粒製劑的生物活性，可能須再修改實驗設計方法加以討論。
　處方F-3的安定性相較於處方F-1和F-2下稍不穩定，但抗癌效果極佳，故本實驗針對處方F-3稍做修飾加強其安定性，修飾處方安定性結果以處方代號TM-PEG+G(62/5)最為安定，因此以此處方再作體外癌細胞之毒殺能力測試，但實驗結果不如原本處方F-3佳。
另一方面以此處方TM-PEG+G(62/5)再衍伸作細胞保護實驗，以過氧化氫為破壞神經細胞(PC 12)的誘導因子，觀察處方TM-PEG+G(62/5)是否有抑制過氧化氫對神經細胞(PC12 cell)產生之傷害，以達保護作用之活性測試。且由流式細胞儀測試結果得知，處方TM-PEG+G(62/5) 確實有被神經細胞吞入，且依處方濃度越高其吞噬量越明顯。並由流式細胞儀得知藥物確實進入細胞中。

關鍵字

類黃酮 ； 固體脂質微粒

並列摘要

Recently, flavonoids constituets with biological activity, such as anticarcinogens, antioxidatives, have received attention. The purpose of the experiment was to evaluate a series of flavonoids derivatives the anticarcinogens, antioxidant activity, free radical scavenging activity and Neuroprotective Effects against H2O2-Induced Cytotoxicity in PC12 Cells in solid lipid nanoparticles formulation. Stability issues from the particles are evaluated in all kinds of materials composed of solid lipid nanoparticles. In addition, the materials composed of solid lipid nanoparticles were discussed the advantage of target on all kinds of cancer cells in this experiment. All compounds were performed cell toxicology (MTT assay), antioxidant activity and free radical scavenging activity.
From the lectures, the materials consisting of solid lipids have an effect on their encapsulation capacity and controlled release properties. It was correlation between antioxidant activity and anticarcinogens of flavonoids derivatives. Bioactivity in flavonoid-SLN wasn`t expected in antioxidant activity and free radical scavenging activity. It was conjectured that flavonoids were covered in solid lipid nanopaticles. Flavonoid-SLN possessed the high Entrapment efficiency. In my study, Formulation were modified to turn into the more stabe one in Flavonoid-SLN. The modified formulation were tested in neuroprotective effects against H2O2-Induced cytotoxicity in PC12 Cells. The survey of the experiment revealed the modified formulation pocessed the neuroprotective effects. The modified formulation were t examined in flow cytometry. We find the formulation were uptaked into cell.

並列關鍵字

flavonoids ； solid lipid nanoparticles

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