吳郭魚膠原蛋白與虎杖萃取物在發展生醫材料之潛力

膠原蛋白是人體結構組織中含量最豐富的結構性蛋白。適用於傷口敷料，促進傷口修復、防止細菌感染以及快速止血的效果，但由於人與禽畜共通疾病的流行，使得魚類成為膠原蛋白較安全的來源。根據文獻指出中草藥虎杖擁有許多治療功效，可減輕傷口感染，促進傷口癒合。本研究目的是萃取吳郭魚魚皮和魚骨中的膠原蛋白，加上中草藥虎杖萃取物，以體外試驗探討其殺菌、抗發炎及促進纖維母細胞(fibroblast)生長激素分泌的功能。結果顯示：利用掃描式電子顯微鏡 (SEM)觀察兩種不同乾燥方法的膠原蛋白膜結構，熱乾燥的膠原蛋白膜結構平滑無孔洞，冷凍乾燥的膠原蛋白結構整齊多孔洞，可得知膠原蛋白在高溫下容易破壞原有的結構，使其變性。藉由抗菌實驗結果不僅得知以虎杖丙酮萃取物抗菌能力佳，其次為虎杖95%乙醇萃取物，也發現虎杖適合利用極性高的溶劑萃取，其中以丙酮溶劑萃取虎杖有效的抗菌成份效果為最佳。經過細胞存活試驗，發現經過低濃度的虎杖萃取物培養後，可促進纖維母細胞的分裂生長，不刺激巨噬細胞引發發炎反應。更進一步分別進行抗發炎與纖維母細胞生長因子試驗，其結果顯示低濃度虎杖水萃取物能夠有效刺激纖維母細胞生長因子的分泌與減少巨噬細胞生成TNF-α，具有降低發炎的功效。綜觀上述研究，魚膠原蛋白適合採用冷凍乾燥方式製作魚膠原蛋白膜，可保持膠原蛋白結構與性質，降低變性風險。虎杖具有良好的抗菌功效之外，能降低發炎，並有促進纖維母細胞生長因子的生成，未來將可運用於製作照護傷口的敷料上。

關鍵字

魚膠原蛋白；虎杖；抗菌；抗發炎；纖維母細胞生長因子

並列摘要

Collagen is the most abundant in the structure of the human body structural proteins. Apply to wound dressings, wound repair, prevention of bacterial infections and rapid hemostatic effect. But because of the common prevalence of human and livestock diseases, making fish become a safe source of collagen. According to the literature, the Chinese herbal medicine Polygonum cuspidatum has many therapeutic effects, which can reduce wound infection and promote wound healing. In current research, extraction of collagen from tilapia’s skin and bone, coupled with Chinese herbal extracts of P. cuspidatum were used for in vitro experiments to explore their sterilization, anti-inflammatory and promote fibroblast growth hormone secretion function. The results show that with a scanning electron microscope (SEM) observed two kinds of collagen film structures of different drying methods, heating-dry collagen film is smooth structure without holes, freeze-dry collagen film is multi-mesh holes, it can be seen that collagen easily degrades the original structure at high temperatures and denatures it. According to the results of the antibacterial experiment, it is not only known that the antibacterial ability of the water extract of P. cuspidatum is good, followed by the 95% ethanol extract of P. cuspidatum, it was also found that the P. cuspidatum was suitable for the use of polar high solvent extraction, in which the effective antibacterial component of the P. cuspidatum was best extracted by acetone solvent. After the cell survival test, it was found that after a low concentration of P. cuspidatum extract cultured, fibroblasts could be promoted to divide and grow, but had no significant effect on the growth of macrophages and doesn’t stimulate macrophages to trigger an inflammatory response. Further anti-inflammatory and fibroblast growth factor tests were performed. The results showed that the low concentration of P. cuspidatum aqueous extract can effectively stimulate the secretion of fibroblast growth factor and effectively reducing macrophage production of TNF-α has the effect of reducing inflammation. Looking at the above research, fish collagen is suitable for the production of fish collagen membrane by freeze-drying method, which can maintain the structure and properties of collagen and reduce the risk of degeneration. In addition to its good antibacterial effect, P. cuspidatum can reduce inflammation and promote the production of fibroblast growth factor, which will be used in the future to make dressings for wound care.