柱层析法分离白蛋白在肝损伤大鼠中的保护效果研究

Column chromatographyanalysison the protective effect of albuminon liver-injured in rats

目的研究采用柱层析工艺直接从血浆中分离的白蛋白在肝损伤大鼠中的保护效果。方法将血浆按1∶4比例稀释后,按照柱层析工艺G25-DEAE-CM-S200获得白蛋白纯品。将其分为A组(5mg/mL)、B组(2mg/mL),另取市售低温乙醇法制备的白蛋白为C组(5mg/mL)。3组按尾静脉注射方式分别对肝损伤大鼠模型进行给药,并以正常组(未造模)及模型组(造模未给药)为参照,比较给药后大鼠肝体系数差异、生理学指标表达水平及肝脏病理损伤程度,以验证2种工艺对蛋白纯度和肝损伤保护效果的影响。结果柱层析工艺最终获得终浓度5mg/mL、纯度98%以上白蛋白;A、B、C3组均能减少血清中谷丙转氨酶的含量,并提高血清中白蛋白含量(P<0.05),但A组保护效果优于B、C组。病理结果进一步证实,A组大鼠仅轻微肝纤维化,B、C组大鼠肝纤维化程度较A组严重。结论不同分离工艺得到的白蛋白,对于小鼠肝损伤程度均有一定缓解,其中以柱层析法所得白蛋白效果较显著。

Objective To evaluate the protective effect of directalbuminchromatographic separation on rats with liver injury.Methods Pure albuminswas obtained by the chromatography standard G25-DEAE-CM-S200 froma 1 ∶4 diluted plasma sample. The albumins were divided into group A( 5 mg/m L) and group B( 2 mg/m L).Albumins prepared by conventional low-temperature ethanol method was assigned as group C( 5 mg/m L).The 3 groups were injectedby a tail vein injection model into liver-injuredrats. Meanwhile a normal group( nom-modeled) and a model group( modeled butnot injected) were set up asreferences. Differences in physiologic expression and pathological liver injury were compared to verify the effects of different treatment on protein activity.Results The chromatography approach obtained a final albumin concentration of 5 mg/m L with more than 98% purity.Group A,B and C could all reduce serum alanine aminotransferase content,and increase serum albumin( P<0. 05). Group A showed superior protectiveeffect than group B and C. Pathology results confirmed that group A rats suffered only minimal hepatic fibrosis. Conclusion The albumin obtained from different separation technology has some relief for the degree of liver injury in mice.Chromatography-separated albumin showed superior effect among all methods.