PLA-O-CMC纳米粒子培养的猪肝细胞大鼠腹腔内移植后凋亡相关蛋白的变化  

The changes of expression of apoptosis-related proteins of the transplantated hepatocytes after intraperitoneal transplantation in rats using porcine hepatocytes cultured with polylactic acid-O-carboxymethylated chitosan nanoparticles

作　　者：陈钟[1] 汤飞[1] 蔡鸿宇[1] 官伟军[1] 

机构地区：[1]南通大学附属医院肝胆外科,江苏南通226001

出　　处：《中国普通外科杂志》2008年第11期1088-1092,共5页China Journal of General Surgery

基　　金：江苏省自然科学基金资助项目(BK200544);南通大学创新基金资助项目(CX0301)

摘　　要：目的探讨聚乳酸-O-羧甲基壳聚糖(PLA-O-CMC)纳米粒子培养的猪肝细胞大鼠腹腔内移植后移植肝细胞凋亡相关蛋白Bcl-2,Bax和Fas表达的变化。方法原位胶原酶循环肝灌注法分离猪肝细胞。D-氨基半乳糖腹腔内注射制作大鼠急性肝衰竭模型。随机分为2组,48 h后分别将5 mLⅠ型胶原凝胶包埋的PLA-O-CMC纳米粒子黏附培养24 h的猪肝细胞(A组),5 mLⅠ型胶原凝胶固定培养24 h的猪肝细胞(B组)移植至急性肝衰竭大鼠腹腔内。移植量均为5.0×107个肝细胞。采用免疫组化法检测移植后3 d内移植肝细胞Bcl-2,Bax和Fas的表达情况。结果移植后1,2,3 d,A组Bcl-2阳性细胞百分数明显高于B组(P<0.05),Bax和Fas阳性细胞百分数明显低于B组(P<0.05)。结论PLA-O-CMC纳米粒子能上调移植猪肝细胞的抗凋亡蛋白的水平和下调凋亡蛋白的水平,具有良好的抗凋亡作用。Objective To investigate the changes of expression of apoptosis-related proteins Bcl-2, Bax and Fas of the transplantated hepatocytes after intraperitoneal transplantation in rats using porcine hepatocytes cultured with polylactic acid-O-carboxymethylated chitosan nanoparticles. Methods Recirculating collagenase perfusion method was used to isolate porcine hepatocytes. At 48h after D-galactosamine intraperitoneal injection, 5 mL polylactic acid-O-carboxymethylated chitosan nanoparticles-cuhured porcine hepatocytes embedded in collagen gel （ group A ） and 5 mL porcine hepatocytes embedded in collagen gel （ group B ） were transplanted into peritoneal cavity of SD rats, respectively. The amount of transplanted porcine hepatocytes was 5.0 × 10^7 hepatocytes in two groups. The expression of Bcl-2, Bax and Fas in transplanted hepatocytes three days after transplantation was detected with immunohistochemistry. Results At 1 , 2 and 3 d after transplantation, the rates of Bcl-2 positive ceils in group A was significant higher than those in group B, whereas the rates of Bax and Fas positive cells in group A were significant lower than those in group B （ P 〈 0. 05 ）. Conclusions Polylactic acid- O- carboxymethylated chitosan nanoparticles can upregulate antiapoptosis protein expression and downregulate apoptosis proteins expression of xenotransplanted porcine hepatocytes inrats, and the anti-apoptosis effect is better.

关 键 词：肝细胞/移植 异种肝细胞移植 肝功能衰竭 急性 纳米粒子 细胞凋亡 

分 类 号：R617[医药卫生—外科学]