不同剂量不同载体不同途径的肝再生增强因子重组质粒抗大鼠肝纤维化疗效比较  

Comparison of different doses,carriers and introducing routes of augmenter of liver regeneration recombinant plasmids in their anti-fibrosis effects in Wistar rats

作　　者：李青[1] 刘殿武[1] 肖永红[1] 刘辉[1] 何海艳[1] 

机构地区：[1]河北医科大学公共卫生学院流行病学教研室,石家庄050017

出　　处：《第三军医大学学报》2006年第18期1833-1836,共4页Journal of Third Military Medical University

基　　金：河北省自然科学基金资助项目(302489)~~

摘　　要：目的筛选肝再生增强因子重组质粒基因治疗时较优的载体、途径和剂量及三者的最佳组合。方法制备肝纤维化大鼠模型,L9(33)正交设计分组,用3种不同的载体(裸基因、脂质体、多聚物)携带不同量(50、100、200μg/kg)的肝再生增强因子重组质粒,采用3种给药途径(肌肉注射、静脉注射、腹腔注射)对模型大鼠进行干预,通过肝组织病理学、肝纤4项等指标来判断疗效并进行对照。结果剂量因素:200μg/kg组纤维化半定量计分和肝纤4项综合得分明显低于50μg/kg组和100μg/kg组,但50μg/kg组和100μg/kg组的结果无显著性差异。载体因素:裸基因组、脂质体组和多聚赖氨酸组3组之间的纤维化半定量计分均无显著性差异。脂质体组和多聚赖氨酸组的肝纤4项综合得分均明显低于裸基因组。途径因素:肌肉注射组、静脉注射组和腹腔注射组3组之间的纤维化半定量计分均无显著性差异。腹腔注射组的肝纤4项综合得分明显高于肌肉注射组和静脉注射组。结论200μg/kg的肝再生增强因子重组质粒的抗大鼠免疫性肝纤维化的疗效优于100μg/kg和50μg/kg;由脂质体携带DNA和由多聚物携带DNA的疗效明显优于裸DNA。肌肉注射和静脉注射的疗效明显优于腹腔注射。由脂质体或多聚物携带、静脉或肌肉注射200μg/kg的肝再生增强因子重组质粒,能取得较好的抗大鼠免疫性肝纤维化的疗效。Objective To select an optimal combined set of carrier, introducing route and dose of recombinant plasmid of augmenter of liver regeneration （ALR） for ALR gene therapy. Methods A total of 40 Wistar rats received intraperitoneal porcine serum administering to develop the immune hepatic fibrosis, and then were divided into 9 groups with orthogonal design L9 （33 ） , each representing one time of test. Hepatic fibrosis rats were given 3 different doses （50, 100, and 200 g/kg） pcDNA3-ALR recombinant plasmids carried by 3 different carriers [ naked plasmid, liposomes and glyco-poly-L-lysine （GPLL） ] through 3 different introducing routes （intramuscular, intravenous and intraperitonea] injection）. The effects were estimated and compared with histopathological changes of liver tissue and 4 fibrosis serum markers, such as hyaluronicacid （HA） , precollagen Ⅲ （ PC Ⅲ ）, collagen Ⅳ （ Ⅳ C ）, and laminin （ LN ）. Results The semiquantitative scoring system （SSS） of liver tissue and the general scoring of 4 fibrosis serum markers were significantly low in 200 g/kg group compared with the other 2 dose groups, but no significantly difference between these 2. No significant difference in the SSS was found among naked gene, liposomes and GPLL group. The general scoring of 4 fibrosis serum markers were lower in liposomes and GPLL group than naked gene group but no significantly difference between liposomes and GPLL group. There was no significantly difference in the SSS among intramusclar,intravenous and intraperitoneal injection group. The general scoring of 4 fibrosis serum markers was higher in intraperitoneal injection group than in other 2 routes, but no significant difference was seen between the later 2. Conclusion The anti-fibrosis effects of 200 g/kg ALR recombinant plasmid are more adaptive than 100 g/kg and 50 g/kg. The liposomes and GPLL are more excellent carrier than naked gene. The intramusclar and intravenous injection are more appropriate gene delivery route than intrape

关 键 词：肝纤维化 肝再生增强因子 基因治疗 脂质体 多聚赖氨酸 正交设计 

分 类 号：Q78[生物学—分子生物学] R575.3[医药卫生—消化系统]