澤蘭素在空氣污染懸浮微粒誘發人類角質細胞發炎過程中之抗發炎及抗氧化機轉

The anti-inflammatory and antioxidant mechanisms of eupafolin on the factor of air pollution: “Particulate Matter (PM)” induced human keratinocytes inflammation

林資展(Zih-Chan Lin)

指導教授 ： 顏峰霖

高雄醫學大學/藥學院/香粧品學系碩士班/碩士(2014年)

摘要

空氣污染已知為人類健康危害的重要環境因子之一，其包含之懸浮微粒更能引發全身性發炎相關的疾病。本論文主要針對空氣污染懸浮微粒引起人類最大器官，皮膚，之致病機轉探討。研究發現，人類角質細胞(HaCaT)受到懸浮微粒刺激能誘導大量環氧化酶二型 (COX-2) 發炎蛋白表現及前列腺素E2生成，這些發炎反應均能受到不同訊號分子抑制劑所阻斷，包括: 抗氧化劑(DPI、APO、NAC)、MAPKs (SB202190、SP600125、U0126) 及 轉錄因子 NF-κB (Bay117082)或是利用專一性更高的 siRNAs ( p47與NOX2) 轉殖技術等，均能由西方墨點法、反轉錄聚合酶連鎖反應、酵素免疫分析及細胞內活性氧化物分析等實驗結果證實。此外，抗氧化劑亦能降低懸浮微粒誘發人類角質細胞p47的活化、細胞內活性氧化物質的生成及多種蛋白質激酶的磷酸化，包括ERK1/2、p38與JNK1/2。進一步，藉由核質分離技術發現調控環氧化酶二型之轉錄因子NF-κB的活化可受到ERK1/2、p38與JNK1/2等蛋白質激酶的影響，其中又以p38路徑的調控為最主要。透過上述實驗結果推測，懸浮微粒在人類角質細胞上可經由NOX2/p47/ROS/MAPKs/ NF-κB路徑誘發環氧化酶二型的大量表現。最後，本研究進一步加入由鴨舌癀植物純化出來的一種類黃酮化合物，澤蘭素，並探討澤蘭素對懸浮微粒刺激人類角質細胞引起上述病理機轉的影響，實驗結果證實澤蘭素
能減少細胞內活性氧化物的產生亦能抑制MAPKs及NF-κB的磷酸化活化，進而降低環氧化酶二型表現和前列腺素E2之生成。綜合以上研究結果，本論文發現澤蘭素可以有效地降低空氣污染懸浮微粒刺激人類角質細胞引起的氧化壓力及發炎，期望未來澤蘭素可添加在化妝保養品中，做為抗空氣污染化妝品活之活性添加物。

關鍵字

澤蘭素 ； 抗皮膚發炎 ； 抗氧化 ； 抗空氣污染化妝品

並列摘要

Air pollution is known to be an important environmental factor of the human health risk, the particulate matter (PM) in air pollution can cause inflammatory related systemic diseases. This thesis is to investigate the different pathogenesis mechanisms participating in PM-induced the largest organ of human body, skin. This study found that PM significantly increased prostaglandin E2 (PGE2) production associated with increased Cyclooxygenase-2 (COX-2) expression in human keratinocytes (HaCaT). This effect was blocked by inhibitors of several signaling transduction molecules including antioxidants (DPI, APO, NAC), MAPKs (SB202190, SP600125, U0126) and nuclear factor-κB (Bay117082) or using the specific siRNAs (p47 and NOX2), all results will show by the methods of western blot, reverse transcription-PCR, intracellular reactive oxygen species assay and ELISA. Moreover, pretreatment with antioxidants led to reduced PM-induced phosphorylation of ERK1/2, JNK1/2 and p38. Further, using cell fraction isolation to analyze HaCaT cell nuclear protein found that MAPK inhibitors attenuated PM-induced increase in the phosphorylation of NF-κB transcription factor and the p38 seems to be the main pathway to regulate. Our results suggest that in HaCaT cells, PM-stimulated COX-2 protein expression and PGE2 production might mediate through NOX2/p47/ROS/MAPKs/ NF-κB pathway. In addition, the study also using eupafolin, a major active component found in the methanol extracts of Phyla nodiflora, has been used to treat inflammation of skin. We examined its effects on COX-2 expression in PM-treated HaCaT cell. Our results reveal a mechanism for anti-inflammatory and anti-oxidative effects of eupafolin that involved inhibition of PM-induced ROS generation, suppression of MAPKs phosphorylation, activity of NF-κB and attenuated COX-2 expression leading to reduced production of prostaglandin E2 (PGE2). In conclusion, the present study suggested that eupafolin can add into cosmetic formulation for claiming the“anti PM cosmetic” through antioxidant and anti-inflammation activity.

並列關鍵字

eupafolin ； anti skin inflammation ； antioxidant ； anti PM cosmetic

參考文獻

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