积雪草提取物对小鼠脾淋巴细胞凋亡的影响

摘要

AIM:To isolate the active compounds of Centella asiatica and to investigate their effects on the apoptosis of mouse spleen lymphocytes. METHODS: Cold soak, polar extraction, silica gel column chromatography and Sephadex LH-20 column chromatography were performed to isolate the active compounds from Centella asiatica, which were identified by the assays of scavenging activity on 1, 1-diphenyl-2-picrylhydrayl radical ( DPPH·) and hydroxyl radical(OH·). Flow cytometry was performed to observe the effects of Centella asiatica extracts on the apoptosis of spleen lymphocytes in mice. RESULTS: Four extracts (Bl, B2, B3 and B4) were obtained by the separating procedure. Among them, the extract B4 exerted a potent scavenging effect on both DPPH·and OH·. Quercetin was identified from the extract B4 by HPLC analysis. The mouse spleen lymphocytes with extract B4 treatment showed a significant rise in the level of apoptosis compared with control group. CONCLUSION: Centella asiatica extract B4 induces apoptosis of spleen lymphocytes, and quercetin is the major substance of extract B4.%目的:分离纯化积雪草活性成份,并观察其对小鼠脾淋巴细胞凋亡的影响.方法:采用乙醇浸提、极性萃取、硅胶柱层析和Sephadex LH-20凝胶柱层析分离纯化积雪草活性成份,通过评价不同阶段积雪草提取物清除1,1-二苯基-2-苦基苯肼自由基(DPPH·)和抑制羟自由基(OH·)能力,筛选积雪草活性成份.采用流式细胞术观察积雪草提取物对小鼠脾淋巴细胞凋亡的影响.结果:(1)积雪草分离纯化后得到提取物B1、B2、B3和B4,其中提取物B4清除DPPH·及抑制OH·的能力最显著.经HPLC分析,积雪草提取物B4主要成分为槲皮素.(2)与对照组相比,积雪草提取物B4作用组小鼠脾淋巴细胞凋亡率显著增加.结论:积雪草提取物B4能诱导小鼠脾淋巴细胞凋亡,其主要成份可能为槲皮素.