靈芝多醣體對內毒素處理小鼠及人類主動脈平滑肌細胞之介白素-1表現的影響及其機轉

The effects of Ganoderma lucidum polysaccharides regulated Interleukin-1 expression in Lipopolysaccharides-treated mice and human aortic smooth muscle cells and its mechanisms

林佳穎(Chia-Ying Lin)

指導教授 ： 陳玉怜

國立臺灣大學/醫學院/解剖學暨生物細胞學研究所/碩士(2008年)

https://doi.org/10.6342/NTU.2008.01256

摘要

Ganoderma lucidum (Reishi; 靈芝) 長久以來被使用做保持健康長壽的漢方藥物，研究發現靈芝中有許多具有生物活性的成份，進而調控身體生理功能，目前已知靈芝有抗腫瘤以及免疫調節的作用。F3 (Fraction 3; EORP) 是從靈芝中萃取出的水溶性多醣體，研究證實，F3 可促使人類巨噬細胞表現細胞素 (cytokines)，如：介白素-1 (interlukin-1β)，增加體內免疫系統功能且增強抵抗力。動脈硬化屬於慢性發炎疾病，細胞素在發炎反應中扮演重要角色，因此本實驗主要探討利用 F3 處理，是否影響 LPS 刺激人類主動脈平滑肌細胞 ICAM-1 及 IL-1 的表現及其相關機制。
實驗結果顯示：以 F3 處理人類主動脈平滑肌細胞，可抑制細胞受到 LPS 刺激所產生的 ICAM-1 表現， 免疫細胞化學染色也觀察到相同結果，並且 F3 處理可降低因 LPS 所引發的單核球沾黏作用；然而，F3 卻可增加人類主動脈平滑肌細胞受 LPS 刺激所表現的 proIL-1 和 IL-1ra，且可促使細胞外 IL-1ra 表現量增加。此外，F3 可調控人類主動脈平滑肌細胞內因 LPS 刺激活化的 MAP kinases (ERK1/2、JNK 及 p38)，F3 可抑制 ERK1/2 的活化，但會促進 JNK 及 p38 的活化。加入 MEK1 抑制劑 (PD98059) 後，以 LPS 刺激發現可抑制 ICAM-1 表現；加入 JNK 抑制劑 (SP600125) 可抑制因 LPS 刺激所表現的 proIL-1 和 IL-1ra。動物實驗方面，F3 與 LPS 短期刺激下，可降低 C57BL/6 小鼠體內 IL-1β和 IL-1ra 表現量；長期刺激下，F3 可降低因 LPS 刺激所表現的 IL-1β，卻可增加 IL-1ra 的表現量，利用 免疫組織化學染色觀察胸主動脈管壁 IL-1β和 IL-1ra 表現，發現也有相同結果。
由以上實驗結果推論：F3 可影響人類主動脈平滑肌細胞內 ERK1/2、JNK 磷酸化，而調控因 LPS 所誘發的 ICAM-1、proIL-1 和 IL-1ra ，並且降低發炎細胞 (如：單核球) 的沾黏，降低發炎細胞引發的組織發炎反應，F3 的餵食也可影響當動物體受 LPS 刺激時 IL-1 的表現，實驗證明 F3 可保護人類主動脈平滑肌細胞受 LPS 刺激後的發炎反應，降低血液中發炎細胞沾黏導致的發炎作用，也可透過調控動物體受 LPS 刺激後 IL-1 表現量的改變，以達到保護動物體避免受 LPS 感染而引發的血管病變。

關鍵字

靈芝多醣體 ； 介白素-1 ； 人類主動脈平滑肌細胞

並列摘要

Ganoderma lucidum (Reishi, Ling-Zhi), a popular home remedy, has been known for its beneficial activities in human health and longevity for centuries. Accumulated studies attempting to understand the role of Reishi in regulating various body functions revealed that the crude or purified components of Reishi extracts possess anti-tumor and immunomodulating activities. A glucan-containing extract of Reishi-derived polysaccharides (EORP; F3) in human macrophages exerts immuno-modulating activities by stimulating the expression of interleukin-1β (IL-1β), a key mediator in chronic vascular inflammatory response. In the present study, we used F3 to test its functional role in LPS-treated smooth muscle cells, which play a key role in atherogenesis and restenosis. LPS treatment increased intercellular cell adhesion molecule-1 (ICAM-1) expression in a time- and dose-dependent manner in human aortic smooth muscle cells (HASMCs). Pretreatment with F3 significantly suppressed the LPS-induced expression of ICAM-1, and the effect was mediated by the inhibition of ERK phosphorylation. F3 also significantly reduced the binding of the human monocytic cell line, U937, to LPS-treated HASMCs. In addition, F3 could regulate the expression of IL-1βand IL-1 receptor antagonist (IL-1 Ra) in LPS-treated HASMCs as well as in plasma and vascular wall in LPS-treated C57BL6 mice by ELISA and Western blotting. These results suggest that F3 may play important roles in the protection of atherosclerosis and inflammatory responses.

並列關鍵字

Ganoderma lucidum ； polysaccharides ； Interleukin-1 ； human aortic smooth muscle cells

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