β-Lapachone加速皮膚燙傷傷口癒合

β-Lapachone Accelerates the Recovery of Burn-Wounded Skin

傅詩宸(Shih-Chen Fu)

指導教授 ： 盧國賢

國立臺灣大學/醫學院/解剖學暨生物細胞學研究所/碩士(2009年)

https://doi.org/10.6342/NTU.2009.02578

摘要

β-lapachone 為一種由南美洲Lapacho tree 樹幹中萃取出來的物質，我們實驗室已證實β-lapachone 具有促進老鼠皮膚傷口癒合的能力。本實驗採用C57BL/6鼷鼠和巨噬細胞株 (RAW264.7 cells) 分別進行活體及細胞培養的研究，探討β-lapachone 對皮膚燙傷傷口癒合以及巨噬細胞增生與釋放生長因子能力的影響，實驗結果如下：(1) 動物實驗結果顯示：經內含100μM β-lapachone 的凡士林軟膏處理後，實驗組老鼠燙傷傷口 (6mm 直徑圓形，100℃，60 秒) 癒合的速度較僅敷上凡士林軟膏之控制組快。在燙傷後14 天，實驗組老鼠燙傷傷口直徑已縮為原來傷口長度的5.4%，但控制組老鼠傷口長度仍為原來的40%，具顯著差異；(2) 蘇木紫-伊紅染色和螢光免疫組織化學染色結果顯示：經100μMβ-lapachone 處理後的燙傷傷口，其組織切片的表皮層、真皮層和皮下組織都較快恢復原有的結構並出現較多的巨噬細胞；(3) 細胞培養的結果顯示：巨噬細胞在0.5μM β-lapachone 處理後24 小時，細胞增生比例為127.9%，與控制組相比，具顯著性差異，且巨噬細胞株在0.5μM β-lapachone 處理後3~24 小時，與同時間未經藥物處理的控制組相比較，細胞增生比例為105% ~ 117%；(4) 酵素免疫分析法結果顯示：先以10ng/ml TNF-α 處理5 分鐘、再加入0.5μM β-lapachone 的處理組可以使巨噬細胞在短時間內釋放最多表皮生長因子 (EGF)，與控制組相比，在15 和30 分鐘時達到顯著差異；但僅加入0.5μM β-lapachone 的處理組可以使巨噬細胞釋放最多血管內皮生長因子 (VEGF)。
綜合以上結果顯示：β-lapachone 可加速燙傷傷口癒合的過程，可能的機制為透過增加巨噬細胞的增生，使其釋放較多的表皮生長因子和血管內皮生長因子，進而達到促進燙傷傷口癒合的目的。

關鍵字

巨噬細胞 ； 傷口癒合

並列摘要

β-lapachone is a quinine of lapachol extracted from the bark of lapacho tree. Our recent findings demonstrated that punched skin wounds of mice healed much faster in β-lapachone treated group than in the control group (Kung et al., 2008). In this study, C57BL/6 mice were used to investigate the effects of β-lapachone on burn-wounded skin. Cell studies were done by using RAW264.7 cell line as a model for scrutinizing the effect of β-lapachone on proliferation and secretion of growth factors of macrophages. In vivo experiments revealed that wounds treated with 100μM β-lapachone recovered faster then those treated with control ointment. H&E and immunohistochemistry staining showed that wounds treated with ointment containing 100μM β-lapachone recovered faster in its epidermis, dermis, and connective tissues, and more macrophages appeared than those treated with control ointment. By MTT assay, We found that after being treated with 0.5μM β-lapachone for 24 hours, the proliferation ratio of RAW264.7 cells raised to 127.9%. The results of time course (3hr~24hr) revealed that the proliferation ratio of 0.5μM β-lapachone-treated RAW264.7 cells raised from 105%~177%, indicating that β-lapachone has reached its maximal effect within 3 hours. The effects of TNF-α and β-lapachone on the secretion of growth factors by macrophages were also analyzed by ELISA. Results indicated that the secretion of EGF by macrophages treated with “5min 10ng/ml TNF-α pretreatment + 0.5μM β-lapachone” was the highest among all five different treatments, and significance was reached at 15 and 30 minutes compared with control. In contrast, the secretion of VEGF by macrophages treated with 0.5μM β-lapachone alone was the highest.
The results obtained from the present study showed that β-lapachone might play an important role in accelerating the burned wound healing process by increasing the proliferation of macrophages and enhancing their ability to secrete EGF and VEGF.

並列關鍵字

β-Lapachone ； EGF ； VEGF

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