靈芝蛋白LZ-8促進傷口癒合研究

LZ-8是一種來自酵母菌重組而成的免疫調節蛋白。過去的研究發現LZ-8可以活化人類T細胞、巨噬細胞、樹突細胞進而造成多種細胞激素（cytokines）及生長因子的產生。然而在傷口癒合早期,急性免疫反應佔有重要角色,其中包含嗜中性球的聚集以及免疫細胞的活化等。因此推論LZ-8與加速傷口癒合具有相關性。我們研究顯示連續七天在C57BL/6小鼠傷口上塗抹3.5 mg LZ-8具有較佳的傷口閉合。經由分析傷口面積,塗抹3.5 mg LZ-8的小鼠在第一天減少近40%的傷口面積。而控制組只有減少10%的傷口面積。因此我們認為LZ-8確實會加速傷口癒合。之後利用10 μg/ml LZ-8處理人類肺纖維母細胞在24小時候有較佳的移動活性,而處理10, 50, 100, 200, 500 μg/ml LZ-8後的人類肺纖維母細胞在24小時候相較於控制組有很好的細胞覆蓋率。在細胞增生測試中,利用2 μg/ml的LZ-8處理有最高的細胞存活率。利用飛行時間二次離子質譜儀分析其350 mg/ml LZ-8中離子含量,發現鈣離子及鎂離子含量很低,因此我們認為其傷口癒合效果與靈芝中的高金屬離子含量無關。結果顯示LZ-8 不管在動物實驗或細胞實驗中都具有提升傷口癒合的效果。

關鍵字

靈芝蛋白傷口癒合

並列摘要

Ling Zhi-8 (LZ-8) derived from yeast recombination is considered as immunomodulatory protein. It has been demonstrated that LZ-8 can activate varieties of immune cells, leading to increased expression and production of several cytokines and growth factors. Those cytokines and growth factors may modulate leukocyte activation and infiltration in the acute inflammatory during wound healing process. Therefore, LZ-8 can potentially affect the recovery from tissue damage. Currently, our studies demonstrated that LZ-8 accelerates wound healing in animal model. The data shows C57BL/6 mice treated with 3.5 mg LZ-8 displayed a significantly smallest wound remaining after treatment days 7. Followed by measurement, analysis and comparison, near to 40 percent improvement in the size of wound recovery in the mice treated with 3.5 mg LZ-8, comparing to the control group without administration of LZ-8. It suggests that LZ-8 can accelerate tissue repairing and healing. We also performed cell migration and wound healing studies. The results demonstrated the human lung fibroblast WI-38 cell treated with 10 μg/ml LZ-8 with various concentration ranged from 10, 50, 100, 200, 500 μg/ml possess better migration activity with improved wound-recovered area compared to control group after 24 hour treatments. In the cell proliferation assay, WI-38 cell treated with 2μg/ml displayed significantly higher cell viability than the control group. To investigate the role of calcium and magnesium in the LZ-8 mediated wound healing acceleration, the Time-of-Flight Secondary Ion Mass Spectrometry (ToF-SIMS) analysis was performed. The date reveals that calcium and magnesium played a very limited role in LZ-8-mediated wound recovery. No calcium and magnesium were detected in our LZ-8 formula, suggesting the healing effect in our animal model wasn’t associated with metal ion. In our preliminary data, it suggests that LZ-8 can accelerate the healing process.

並列關鍵字

LZ-8 wound healing skin

參考文獻

1. Lin, W.H., Hung, C.H., Hsu, C.I. & Lin, J.Y. Dimerization of the N-terminal amphipathic alpha-helix domain of the fungal immunomodulatory protein from Ganoderma tsugae (Fip-gts) defined by a yeast two-hybrid system and site-directed mutagenesis. J Biol Chem 272, 20044-20048 (1997).

4. Hsu, H.Y., et al. Reishi immuno-modulation protein induces interleukin-2 expression via protein kinase-dependent signaling pathways within human T cells. J Cell Physiol 215, 15-26 (2008).

5. Lin, Y.L., et al. An immunomodulatory protein, Ling Zhi-8, induced activation and maturation of human monocyte-derived dendritic cells by the NF-kappaB and MAPK pathways. J Leukoc Biol 86, 877-889 (2009).

6. Yeh, C.H., Chen, H.C., Yang, J.J., Chuang, W.I. & Sheu, F. Polysaccharides PS-G and protein LZ-8 from Reishi (Ganoderma lucidum) exhibit diverse functions in regulating murine macrophages and T lymphocytes. J Agric Food Chem 58, 8535-8544 (2010).

7. Haak-Frendscho, M., Kino, K., Sone, T. & Jardieu, P. Ling Zhi-8: a novel T cell mitogen induces cytokine production and upregulation of ICAM-1 expression. Cell Immunol 150, 101-113 (1993).

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靈芝蛋白LZ-8促進傷口癒合研究

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