奈米化中草藥對兔子骨髓間葉幹細胞生長影響之研究

本研究以骨髓多能性幹細胞作為評估體外誘導分化模組，探討奈米化中草藥刺激對細胞增生及分化之影響。物質奈米化後有著不同於傳統的特性，奈米化中草藥由於其粒徑小，體表面積增大，使吸收效率增加，因此奈米中草藥產業快速竄起，更被政府列為高值化計畫重點發展項目之ㄧ。將大葉骨碎補、川牛膝、續斷和健步丸複方四種骨傷科中草藥，利用奈米濕式球磨機，依時間長短研磨成不同粒徑大小，再以中藥水萃物做細胞刺激實驗；藉由定量聚合酶連鎖反應 (RT-PCR) 分析，探討骨細胞分化之骨化表現，如骨橋蛋白、骨鈣蛋白。本研究成功地將四種骨傷科中藥利用奈米濕式球磨機，經研磨時間上的比較，藥物粒徑達至300 nm 以下；水萃物以1 mg/mL的濃度作為最高濃度檢測，細胞並未顯示出細胞毒性的現象；再利用其增生參數刺激骨髓間葉幹細胞探討細胞分化，結果發現，皆有促進細胞分化，且時間上和藥效上皆比誘導組來得快且高，對照傳統粒徑的比較上，大葉骨碎補、川牛膝和健步丸複方皆有統計上差異；RT-PCR實驗表現出，四種藥物的刺激所分泌表現出的骨橋蛋白皆比誘導組顯著，而骨鈣蛋白比較於誘導組皆來得緩慢；經由藥物刺激又加上誘導劑的組別表現出有加乘的效果。本研究對中草藥微小化做效益和安全性及細胞刺激之評估，其釋放成分高低和藥物大小具有正向性的關係，並與傳統藥物做比較，提供未來進一步推展以動物實驗模式之參考，開發出對骨傷科更具療效的奈米化中草藥。

關鍵字

川牛膝；續斷；骨髓間葉幹細胞；大葉骨碎補；定量聚合?連鎖反應；健步丸；奈米化

並列摘要

There are many kinds of new properties of nano-materials comparing with the traditional one, so more and more people are interested in the nano-Chinese herb especially for its high efficient absorption. The therapeutic mechanisms of the nano-Chinese herb are still unknown. The purpose of this study is to compare the effects of nano and traditional Chinese herb on bone marrow stromal (BMS) cells proliferation and differentiation in vitro. Four kinds of bone traumatology Chinese herb medicine (Davallia divaricata HAYATA, Cyathula officeinalis KUAN, Dipsacus asper WALL and Jian Buh Wan) were grinded into different sizes according to different grinding period by the nano-scaled wet type ball machine and then the water extractions of them were tested on the cells. After the treating, the osteogenic marker (osteopontin and osteocalcin) during the bone cell differentiation would be evaluated by the quantity-real time polymerase chain reaction (RT-PCR). The results showed that there was no toxic effect of water extracting nano-Chinese herbs (particle size below 300 nm) when the concentrations were lower than 1 mg/mL on BMS cells. The proliferation and differentiation of BMS cells would be increased significantly at certain concentration of nano-Chinese herb. The cell differentiation of nano-scale herb group would be turn on earlier and stimulating effects were higher than the differentiation induced reagent group and the traditional-Chinese herb group. The secretion of osteopontin stimulated by the nano-Chinese herb group was increased significantly than the differentiation induced reagent group, but the response of the secretion of osteocalcin would be late in the nano-Chinese herb group. These effects of the nano-Chinese herb on BMS cells would be more apparently accompanying with the adding of the differentiation induced reagent group at the same time. The effects of the nano-scale Chinese herb on BMS cells are size and dose dependent. The results of this research will be very useful for the size and dosage suggestions in the animal test and to develop a more effective nano-scale Chinese herb type on the bone traumatology.

並列關鍵字

nano-scaled ； bone marrow stromal cells ； Cyathula officeinalis KUAN ； Dipsacus asper WALL ； Jian Buh Wan ； Davallia divaricata HAYATA ； quantity-real time polymerase chain reaction

參考文獻

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奈米化中草藥對兔子骨髓間葉幹細胞生長影響之研究

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