陳皮素對內皮細胞引發發炎反應之保護角色的探討

Evaluation of the Protective Effects of Hesperetin on Endothelial Cells under Inflammatory Stimulation

王秋雲(Chiu-Yun Wang)

指導教授 ： 何豐名 ； 翁清松

中原大學/工學院/醫學工程研究所/碩士(2006年)

https://doi.org/10.6840/cycu200600406

摘要

血管內皮細胞的功能在於調節血管功能並維持其恆定性，所以內皮細胞的功能異常被認為是血管病變的開始。當內皮細胞受傷時，大量分泌出的附著因子會增加單核球(Monocytes)對內皮細胞吸附的能力，進而移行(Migration)到內皮細胞內行使發炎反應，而心血管疾病中的動脈粥狀硬化，即是一種與血管功能異常相關的慢性發炎反應疾病。在許多形式的壓力下，有些細胞會誘導HO-1的表現去行使細胞層級的防衛機制，有效的調控HO-1的分泌可以應用在治療因發炎反應所引起的疾病。陳皮素(Hesperetin)是屬於生物類黃酮-柑橘黃酮中的一種，廣泛存在於芸香科植物和中草藥植物中，而生物類黃酮一族一直是廣泛用來做研究、保健、預防疾病生成的一群天然物。
因此在本研究中，利用TNF-α當作原發炎性刺激物，刺激人類臍帶內皮細胞(HUVECs)作為發炎反應的模式，並添加Hesperetin不同濃度(3μM、10μM、30μM與50μM)觀察此天然中草藥物Hesperetin是否可以保護細胞並降低受刺激後的發炎現象，如觀察內皮細胞的細胞存活率(MTT assay)、分泌IL-8分子和ICAM-1分子的表現量及THP-1附著在內皮細胞上之能力，以及啟動細胞自我防衛層級HO-1分泌的效力。
由實驗結果得知，在Hesperetin濃度為3μM、10μM、30μM下有保護細胞的生長，且在30μM下具有上升細胞受TNF-α(50ng/ml)傷害的細胞存活率。並且3μM、10μM與30μM濃度下皆可抑制TNF-α(50ng/ml)刺激細胞引起之IL-8分子的表現量。在此相同濃度下(3μM、10μM、30μM)，亦能抑制由TNF-α(50ng/ml)所引起的ICAM-1的分泌，並且有濃度的相依性。在THP-1附著的實驗裡，Hesperetin(3μM、10μM、30μM)皆能抑制單核球(THP-1)吸附在內皮細胞上，推論可降低血管發炎初期的反應。且能啟動內皮細胞內皮性HO-1的分泌，增加細胞自我防衛層級的表現，可去調控因發炎反應所引起的疾病。
所以，中草藥物-陳皮素(Hesperetin)能有效的抑制單核球附著到內皮細胞上，且可以啟動細胞的自我防禦機制，具有良好抗發炎性的作用，有可能成為新型開發應用於預防動脈粥狀硬化形成的新中草藥物。

關鍵字

陳皮素 ； 細胞間附著因子-1 ； 動脈粥狀硬化 ； 血基質氧化脢-1 ； 介白質-8

並列摘要

Attachment of leukocytes to the vascular endothelium and then migration into the vessel wall are the early events in atherogenesis. HO-1 is a critical protein in the response to oxidative injury. Its main function is associated with the degradation of heme to biliverdin, iron and carbon monoxide. HO-1, which functions as part of a cytoprotective mechanism, is based upon a series of antioxidant activities. Hesperetin belongs to the class of flavonoids called flavanones, which are abundant in citrus fruits. It has been long known that flavonoids possess bioactive potential for their bioavailability, metabolic fate, and health effects. Flavonoids are potent antioxidants in vitro, and therefore one of the main interests in the compounds has involved protection against cardiovascular disease.
In this study, we used the pro-inflammatory cytokine, TNF-α (10ng/ml, 50ng/ml), to induce the inflammatory reactions in human umbilical vein endothelial cells (HUVECs). Different concentrations of Hesperetin (3μM, 10μM, 30μM and 50μM), the extract of Chenpi, were added into HUVECs for 24 hours to study the cells’ proliferation, the expressions of ICAM-1 and IL-8, the ability of THP-1 adhesion on endothelial cells, and the protein expressions of HO-1.
The results indicated that three concentrations of Hesperetin (3μM, 10μM, 30μM) could increase cell proliferations. The Hesperetin could also show the protective affection on the cells under the stimulation of TNF-α. In the different doses of 3μM, 10μM and 30μM, Hesperetin suppressed the cytokine-induced expressions of IL-8. Hesperetin has shown the ability to reduce the cytokine-induced expressions of ICAM-1, the ability of THP-1 adhesion on endothelial cells, and also the ability to induce the protein expressions of HO-1.
Finally, our results suggest that the extract of Chenpi, Hesperetin, has the possibility to be an anti-inflammatory product.

並列關鍵字

Atherogenesis ； Intercellular Adhesion Molecule-1 ； Hesperetin ； Interleukin-8(IL-8) ； Heme Oxygenase-1(HO-1)

參考文獻

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