美洲大蠊抗肝纤维化活性部位的筛选

Screening of anti liver fibrosis active fractions from Periplaneta Americana

目的结合体外活性筛选,对美洲大蠊提取物进行分离、纯化与活性追踪,以便获得抗肝纤维化活性较强部位。方法用5倍量95%,90%,80%,70%,60%,50%的乙醇溶液和水制备美洲大蠊冻干粉,分别命名为美蠊-A(ML-A)、美蠊-B(ML-B)、美蠊-C(ML-C)、美蠊-D(ML-D)、美蠊-E(ML-E)、美蠊-F(ML-F)、美蠊-G(ML-G)。将细胞分为3组:正常组、空白组(不含细胞)和给药组(美洲大蠊冻干粉溶液:1000,250,62.5,15.63,3.91,0.98μg·mL-1)。用MTT法对美洲大蠊不同溶剂提取物进行抗肝纤维化活性筛选,用HP20大孔树脂对筛选得到的活性部位进行分离与纯化,进行体外活性追踪。结果美洲大蠊提取物ML-D对HSC-T6细胞有一定的抑制作用,其48h的IC_50值为389μg·mL^(-1)。分离ML-D得ML-D_1、ML-D_2、ML-D_3、ML-D_4、ML-D_5、ML-D_6和ML-D_7洗脱部位,其中ML-D_5、ML-D_6、ML-D_7及其合并物ML-HB对HSC-T6细胞细胞的生长有良好的抑制作用,48h的IC_50值分别为115.6,116.5,121.8和110.0μg·mL-1。结论从美洲大蠊中提取分离得到活性部位ML-HB对HSC-T6细胞的增殖有较好的抑制作用。

Objective To combine with in vitro activity screening,the extracts of Periplaneta Americana will be separated,purified and traced to obtain the fractions with strong anti-hepatic fibrosis activity. Methods The lyophilized powder of Periplaneta Americana was prepared with 5 times of the amount of 95%,90%,80%,70%,60%,50% ethanol solution and water,named Mei lian-A( ML-A),Mei lian-B( ML-B),Mei lian-C( ML-C),Mei lian-D( ML-D),Mei lian-E( ML-E),Mei lian-F( ML-F),Mei lian-G( ML-G),respectively. The cells were divided into three groups: normal group,blank group( without cells), and administration groups( Periplaneta Americana lyophilized powder solution: 1000,250,62. 5,15. 63,3. 91,0. 98μg·mL-1). The anti-liver fibrosis activity of different solvent extracts of Periplaneta Americana was screened by MTT method. The active fractions were isolated and purified by HP20 macroporous resin,and their activities were tracked in vitro. Results The Periplaneta Americana extract ML-D had a certain inhibition on HSC-T6 cells,its IC50 value was 389 μg · mL-1 at 48 h. The in vitro activities of the seven eluted fractions,ML-D1,ML-D2,ML-D3,ML-D4,ML-D5,ML-D6 and ML-D7 obtained from ML-D,were determined,the results showed that ML-D5,ML-D6,ML-D7 and their combination ML-HB hadstrong inhibitory effects on the growth of HSC-T6 cells,with IC50 values of 115. 6,116. 5,121. 8 and 110. 0μg·mL-1 at 48 h. Conclusion The active fraction ML-HB obtained by extraction and isolation from Periplaneta Americana has a good inhibitory effect on the proliferation of HSC-T6 cell.