龍膽瀉肝湯對紅斑性狼瘡小鼠之調控機制研究(2-2)

全身性紅斑性狼瘡是一種慢性、多器官侵蝕、系統的發炎性，它和免疫系統失調及廣泛性地免疫複合體沈積有關，同時可以見到細胞激素和發炎細胞共同產生的炎症反應。其病理機轉最主要是導因於身體所產生不同種類的自體抗體，以致於造成身體各個系統及器官的功能喪失會危及生命的疾病。龍膽草(Gentianae Radix)是中醫臨床最常用的清熱瀉火劑之一，原為清溼熱及抑制發炎反應，但目前所使用的範圍則越來越廣，但真正在實驗性紅斑性狼瘡的作用機制卻一直缺乏科學性的探討。 本計劃以MRL lpr/lpr 小鼠為自發性的自體免疫疾病動物模式，小鼠在長大的過程中會出現類似人類全身性紅斑性狼瘡的症狀。實驗動物與以餵食龍膽草正丁醇萃取物二週後，收集尿液、血液與腎臟組織進行病理檢驗與免疫分析。收集血清進行抗雙股去氧核糖核酸抗體分析。其次在本研究中，我們嘗試以蛋白質體學的方法來探討對紅斑性狼瘡動物模式有療效的指標蛋白。實驗結果發現，龍膽草正丁醇萃取物可以降低紅斑性狼瘡小鼠腎臟的氧化壓力。其次對改善MRL 小鼠有嚴重的腎膈增生，腎臟組織IgG 染色沉積較輕。蛋白質體研究各組間有變化的標誌蛋白與腎臟滲透壓調控有密切的關係。有趣的是MRL 小鼠呈現明顯的脾臟 腫大的現象，餵食龍膽草正丁醇萃取物的小鼠脾臟明顯縮小 (1.1±0.34 v.s.0.45±0.21 g)，我們分析此作用與脾臟中調節性T 細胞的表現有關(2.0±0.2 v.s.2.9±0.2%)。這項優勢也表現在顯著降低血中Anti-dsDNA (34355±1459 v.s.13445±1250 ng/ml)、TNF-α (9.1±1.4 v.s.1.6±0.2 pg/ml)與IFN-γ (63±.4 v.s. 18±2 pg/ml)的濃度。藉由此研究，我們希望能進一步評估龍膽草具有治療人類全身性紅斑性狼瘡的潛力。
Systemic lupus erythematosus (SLE) is a chronic multisystem disorder of presumed autoimmune origin in which cytotoxic antibodies, or circulating immune complexes, give rise to tissue damage often resulting in end organ disease and even mortality. Gentianae Radix is a herbal medicine with clear shire and possess antiinflammation effects. In the present study we investigate 1-butanol extract of Gentianae Radix immunomodulatory effect on inhibiting autoimmune disease in MRL lpr/lpr mice. MRL lpr/lpr mice treats daily with 1-butanol extract of Gentianae Radix for 2 weeks. We will study the role of immunological effect of Treg and pathological effect of kidney tissue and progression in anti-ds-DNA production, lymphadenopathy after herbal medicine administration. The protein expression profiles in kidney tissue that are associated with the physiological and pathological change during SLE progression will be analyzed by two-dimension gel electrophoresis and image analysis. Histologic analysis of kidney tissue indicated that 1-butanol extract of Gentianae could inhibit the mesangial proliferation that was evident in lupus nephritis. In addition 1-butanol extract of Gentianae could promote SMP30 and MnSOD levels in kidney tissue of MRL mice. In addition, 1-butanol extract of Gentianae activated Treg (2.0±0.2 v.s. 2.9±0.2%) and markedly reduced anti-ds-DNA (34355±1459 v.s. 13445±1250 ng/ml) autoantibody, IFN-g (63±.4 v.s. 18±2 pg/ml), TNF-a (9.1±1.4 v.s. 1.6±0.2 pg/ml) and IgG immune-complex deposition in kidney tissue. The study would fill a long-existing gap in Chinese medicine in the pathophysiological disorder of SLE. It may also lead to the discovery of potentially useful herb drugs for treating SLE in human patients.