三黃瀉心湯經由抑制p38 MAP kinase相關路徑治療內毒素引發之低血壓及肺部傷害

Therapeutic effects of SHXT on endotoxemia induced hypotension and lung injury by its inhibition of p38 MAP kinase pathway

林怡伶(Yi-Ling Lin)

指導教授 ： 羅怡卿

高雄醫學大學/醫學院/醫學研究所/碩士(2004年)

摘要

三黃瀉心湯 (San-Huang-Xie-Xin-Tang, SHXT) 是中國的傳統方劑，其典故出自於金匱要略，由黃芩(Huang Qui)、黃蓮 (Huanglian)及大黃 (Dauhuang) 以1:1:2的方式加以組成。在民間，三黃瀉心湯被廣泛的使用在清除肺熱、抗發炎及驅淤血的治療上，有鑑於此，本研究將進一步探討三黃瀉心湯在動物模式下，是否能治療由內毒素脂多醣體 (Lipopolysaccharides, LPS) 所誘發的全身性發炎反應及肺部損傷，並深入的探討其抗發炎的作用機制。在本實驗中，我們以靜脈注射的方式給予大鼠LPS，並在十分鐘後投予待測藥物，經過五小時後，取其肺及血液，以西方點墨法及酵素免疫分析法來分析肺部及血漿中蛋白質的變化。經由實驗結果顯示，三黃瀉心湯、baicalin 及dexamethasone (DEXA)皆能顯著的減少內毒素血症的死亡率，並具有對抗內毒素所引起的低血壓及肺水腫現象。經西方點墨法證實，三黃瀉心湯、baicalin及DEXA皆能顯著的抑制內毒素在肺組織所引起發炎相關蛋白質p38 MAP (mitogen activated protein) kinase、transforming growth factor-[beta] (TGF-β)、inducible nitric oxide synthase (iNOS)及intracellular adhesion molecule-1 (ICAM-1)的表現。而經由酵素免疫分析法也確認三黃瀉心湯、baicalin及DEXA能顯著抑制內毒素所引起的細胞激素interleukin-1[beta] (IL-1β)、tumor necrosis factor alpha (TNF-α)及Monocyote Chemoattractant Protein-1 (MCP-1)的產生。三黃瀉心湯在本實驗中已經證實具有抗發炎的作用，尤其三黃瀉心湯在TGF-β及ICAM-1的抑制及肺水腫的治療上有著比類固醇更優異的治療效果。由以上的結果發現，三黃瀉心湯其抗發炎機制是透過抑制p38 MAP kinase所主導衍生的發炎路徑，並且經由減少其下游路徑所產生的發炎物質而達到其抗發炎的效果。以上的研究結果，可作為三黃瀉心湯未來在臨床運用上的重要依據。

關鍵字

肺水腫 ； 內毒素血症 ； 三黃瀉心湯 ； 細胞激素 ； 低血壓

並列摘要

San-Huang-Xie-Xin Tang (SHXT) is a traditional Chinese medicinal formula containing Huang Qui, Huanglian and Dauhuang. In this study, SHXT was first examined for its therapeutic effects on LPS-induced systemic inflammatory reaction and lung injury and further to comparison with baicalin and dexamethasone (DEXA). Acute lung injury (ALI) caused by endotoxemia has a high mortality rate and various inflammatory mediators have been associated with in LPS-induced systemic inflammatory reaction and lung dysfunction. During which, p38 MAP kinase appear to play a major role in systemic inflammation and acute lung injury. In the downstream effects of p38 MAP kinase activation involve generation of NO production by increase iNOS, production of cytokines such as IL-1β, IL-8, TNF-α, adhesion factors and TGF-β. NO generated from iNOS in endotoxin shock plays an important role in vascular hyporeactivity and tissue damage through its cytotoxic function. In this study, SHXT, baicalin and DEXA significantly inhibited LPS-induced hypotension and increased survival rates of endotoxic shock. Western blotting analysis indicated that SHXT, baicalin and DEXA significantly inhibited LPS-induced iNOS expression in the lung. TGF-β plays an important role in the development of pulmonary edema during the early phase of acute lung injury. Epithelial injury and extensive fibrosis are correlated with mortality rate and that both may be mediated by activated TGF-β. Our results also indicated that SHXT inhibits TGF-β and ICAM-1 expressions in the lung and decreased edema index. Results from ELISA analysis indicated that SHXT, baicalin and DEXA decreased plasma levels of IL-1β, TNF-α and MCP-1 induced by LPS. In conclusion, SHXT inhibited LPS-induced hypotension and lung edema by its suppressions of cytokine productions such as IL-1β, TNF-α, MCP-1 and inhibitions of TGF-β, ICAM-1 and iNOS expressions, which might mediate by the inhibition of p38 MAP kinase activation. These results suggest that SHXT may be therapeutically useful for endotoxic hypotension and lung injury.

並列關鍵字

cytokine ； p38map kinase ； tnf ； mcp-1 ； endotoxemia ； shxt ； baicalin ； dexamethasone ； lps ； il-1 ； edema index ； pulmonary edema ； inflammation

參考文獻

參考文獻

Adams DM, Mathaway M and Shaw J. (1991) TGF-β induces T-lymphocyte migration in vitro. J Immunol, 147: 609.

Alderton WK, Cooper CE and Knowles RG. (2001) Nitric oxide synthase: structure, function and inhibition. Biochem J, 357: 593-615.

Anderson RR, Holliday RL, Driedger AA, Lefcoe M, Reid B and Sibbald WJ. (1979) Documentation of pulmonary capillary permeability in the adult respiratory distress syndrome accompanying human sepsis. Am Rev Respir Dis, 119: 869–877.

Annane D, Bellissant E, Sebille V, Lesieur O, Mathieu B, Raphael JC, Gajdos P. (1998) Impaired pressor sensitivity to noradrenaline in septic shock patients with and without impaired adrenal function reserve. British Journal of Clinical Pharmacology; 46:589-97.