三黃瀉心湯抑制細胞發炎機制的探討

Evaluation of the Inhibition Mechanisms of Cellular Inflammation by Herbal Medicine San-Huang-Sei-Sin-Tang

吳章瑜(Chang-Yu Wu)

指導教授 ： 翁清松 ； 何豐名

中原大學/工學院/醫學工程研究所/碩士(2007年)

https://doi.org/10.6840/cycu200700859

摘要

單核球細胞經分化後會形成巨噬細胞(Macrophages)；而巨噬細胞是防禦外來感染原的重要防線之一，在人體中的防禦能力佔有很重要的地位，當身體受到外來物質（細菌或其他物質）入侵或感染時，身體中的巨噬細胞會主動因刺激而分泌許多細胞激素與一氧化氮（發炎指標物之一）來保護身體避免傷害，但如果一氧化氮分泌不足或過量，對於周遭細胞、組織、器官都將造成一定的危害。
中藥複方劑三黃瀉心湯（San-Huang-Shi-Sin-Tan；SHSST）-大黃、黃蓮、黃芩經溶劑萃取出的黃酮素的成份為Flavanones的結構異構物，也是一種抗氧化劑，而Flavanones對於細胞過度發炎及腫瘤的抑制，有一定的效果。
本研究首先利用SHSST 對人類單核球細胞（Human monocytic leukemia cell,簡稱THP1)之存活率及分泌一氧化氮的影響，再利用100ng/ml的脂多醣 ( Lipopolysaccharide, LPS )處理THP1，使細胞受脂多醣刺激而分泌大量一氧化氮，再利用添加SHSST的方式，抑制其發炎現象。最後利用西方墨點法證實，一氧化氮降低是因為添加三黃瀉心湯萃取物抑制了誘導型一氧化碳合成酶（iNOS, inducible nitric oxide synthase）。
實驗結果發現加入SHSST後，細胞不會因SHSST產生發炎反應，但細胞細胞經脂多醣處理後，會因添加SHSST而使一氧化氮產生量降低，並且依SHSST濃度增加使抑制能力上升。在蛋白質的結果方面，我們看到了一氧化氮合成酶(nitric oxide synthase, NOS)與第二型環氧化酶( Cyclooxygenase, COX-2 ) 濃度，會因SHSST增加而降低。所以我們大膽推論，適量的SHSST在正常的情況下，不會使細胞受到傷害，也不會引起細胞的發炎反應，在細胞發炎反應中，卻可以有效的抑制過度發炎的情況，經由在蛋白質時間點的表現上可明顯的得知先行抑制第二型環氧化酶反應後再抑制一氧化氮合成酶來降低一氧化氮的生成。

關鍵字

脂多醣 ； 三黃瀉心湯 ； 第二型環氧化? ； 一氧化氮合成? ； 巨噬細胞

並列摘要

Macrophages play a key role in the immune protection system against microorganisms or tumour cells since they partake in the processing of antigens and presentation to lymphocytes, as well as in the phagocytosis and killing of microbes. In this process, high amounts of nitric oxide (NO) and reactive oxygen intermediates are generated contributing to intracellular destructive mechanisms.
While parts of our body are infected with bacteria, macrophages will secrete cytokines to protect us from being injured. However, if the system secretes too much or too little cytokine, it will cause a threat to cells, tissues, or organs.
The compound of flavanones extracted from SHSST（San-Huang-Shi-Sin-Tan）of Chinese herbal medicine is a kind of antioxidant. Furthermore, flavanones can prevent bacteria and tumors from spreading.
Our study applied SHSST on cells, and it made an impact on the Human monocytic leukemia cell（ THP1). In addition, when we treated the THP1 with LPS, it stimulated cells to secrete a large amount of nitric oxide. Besides, we treated lipopolysaccharide (LPS) on THP1 and it stimulated cells to secrete nitric oxide. Also, the study used SHSST to treat the cells and restrained them from being inflamed. According to Western Blot, nitric oxide decreased because naringenin restrained inducible nitric oxide synthase (iNOS) from developing.
In the experiment, our results indicated that the cells would not be inflamed by SHSST. However, after treating LPS stimulation, nitric oxide could be reversed by SHSST. As for the expression of proteins after LPS treatment, the expression of inducible nitric oxide synthase and cyclooxygenase (COX-2) decreased when SHSST was increased. When SHSST was added to the cells, it was able to reduce the amount of iNOS, which diminished the nitric oxide, and COX-2. Therefore, it could be concluded that the cells would not be inflamed or damaged under the proper amount of SHSST.

並列關鍵字

SHSST ； inducible nitric oxide synthase (iNOS) ； lipopolysaccharide ； cyclooxygenase(COX-2) ； macrophages

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