鮑魚菇蛋白 Pcy 對小鼠腹腔巨噬細胞的活化作用

A Protein Pcy Isolated from Pleurotus cystidiosus Induces the Activation of Mouse Peritoneal Macrophages

鍾易蓁(Yi-Chen Chung)

指導教授 ： 許輔

國立臺灣大學/生物資源暨農學院/園藝暨景觀學系/碩士(2017年)

https://doi.org/10.6342/NTU201603265

摘要

鮑魚菇 (Pleurotus cystidiosus) 是臺灣常見菇類，為側耳科 (Pleurotaceae) 側耳屬 (Pleurotus) 真菌，過去文獻指出，鮑魚菇萃取物具有抗高血壓、抗癌細胞增生與降低血糖等多項生物活性，然而卻少有實驗使用純物質進行研究。因此本研究目的以純化鮑魚菇 14 kDa 之蛋白 (Pcy) 為材料，並研究 Pcy 對小鼠腹腔巨噬細胞 (peritoneal macrophage, pMø) 的免疫調節活性與活化路徑。
經實驗結果發現，將 12.5 μg/mL Pcy 與 pMø 共培養，可以活化 pMø 並增強 pMø 吞噬大腸桿菌之活性，增加表面抗原 CD86 與 MHC class II 表現，提高細胞激素 TNF-α 與 IL-1β 分泌量，同時也提高 TNF-α、IL-1β、IL-12p35、IL-12p40、CD38、EGR2 和 IL-10 等細胞激素之基因表現量。另外，Pcy 可以增加 M1 型 CCL3、CCL4 等趨化素基因之表現，以及 M2 型趨化素 CCL17 和 CCL22 基因表現量也會增加。在訊息傳導途徑方面，分別使用 TLR2 與 TLR4 基因剔除小鼠，發現 Pcy 會活化 TLR2-/- 小鼠腹腔巨噬細胞活化產生 TNF-α 與 IL-1β，然而於 TLR4-/- 小鼠之 TNF-α 與 IL-1β 分泌量皆不明顯，因此推測 Pcy 活化小鼠腹腔巨噬細胞之途徑和 TLR4 相關，然而還需進一步研究驗證。由以上實驗結果可知 Pcy 能活化小鼠腹腔巨噬細胞及其表面標誌，提升宿主免疫反應。

關鍵字

鮑魚菇 ； 免疫調節蛋白 ； 小鼠腹腔巨噬細胞 ； 類鐸受體

並列摘要

Abalone mushroom (Pleurotus cystidious) is one of the most popular edible mushrooms in Asia. Previous studies have shown that extracts from P. cystidious possessed several biological activities including antihypertension, antiproliferation of tumor cells and hypoglycemic effect. However, little research put emphasis on purified protein from P. cystidious. In this study, a 14 kDa immunomodulatory protein, Pcy, was purified from P. cystidious, and the effects of Pcy on activation, differentiation and TLR-signaling pathways of mouse peritoneal macrophages were investigated. According to the results, Pcy could induce phagocytic activity, increase the secretion of TNF-α and IL-1β, and enhance the expression of surface markers CD86 and MHC class II of mouse peritoneal macrophages. Pcy could upregulate mRNA expression of TNF-α, IL-1β, IL-12p35, IL-12p40, CD38, EGR2, and IL-10 cytokines. Pcy could also increase mRNA expression of both M1 and M2 type chemokine genes. Finally, TLR2 and TLR4 knockout mice were used to investigate the signaling pathway. Pcy could still induce macrophages to secrete TNF-α and IL-1β from TLR2 knockout mice, but the same activation was not obvious in TLR4 knockout mice. We suggest that the activation of Pcy on macrophage might relate to TLR4 signaling pathway, and more researches should be done for further confirmation. In conclusion, Pcy is an immunomodulatory protein, which could activated macrophages and strengthen the host immunity.

並列關鍵字

Pleurotus cystidiosus ； immunomodulatory protein ； mouse peritoneal macrophage ； Toll-like receptor

參考文獻

江金標. (1973). 鮑魚菇栽培之基礎研究. 中國園藝, 19(6), 410-415.

Aderem, A., & Underhill, D. M. (1999). Mechanisms of phagocytosis in macrophages. Annual review of immunology, 17(1), 593-623.

Akira, S., Takeda, K., & Kaisho, T. (2001). Toll-like receptors: critical proteins linking innate and acquired immunity. Nature immunology, 2(8), 675-680.

Akira, S., Uematsu, S., & Takeuchi, O. (2006). Pathogen recognition and innate immunity. Cell, 124(4), 783-801.

Auffray, C., Sieweke, M. H., & Geissmann, F. (2009). Blood monocytes: development, heterogeneity, and relationship with dendritic cells. Annual review of immunology, 27, 669-692.