积雪草提取物对小鼠脾淋巴细胞凋亡的影响

Effects of Centella asiatica extracts on apoptosis of mouse spleen lymphocytes

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。

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 compound＇s from Centella asiatica, which were identified by the assays of scavenging activity on 1,1-diphenyl-2-picrylhydrayl radical （ DPPI-I ~ ） and hydroxyl radical （ OH ~ ）. Flow cytometry was performed to observe the effects of Centella asiatica extracts on the apoptosis of spleen lym- phocytes in mice. RESULTS： Four extracts （B1, B2, B3 and IM） 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 ex- tract IM by HPLC analysis. The mouse spleen lymphocytes with extract Bd treatment showed a significant rise in the level of apoptosis compared with control group. CONCLUSION： Centella asiatica extract Bd induces apoptosis of spleen lympho- cytes, and quercetin is the major substance of extract I34.