聚乳酸-O-羧甲基壳聚糖纳米粒子对异种移植猪肝细胞凋亡的抑制作用  

Effect of polylactic acid-O-carboxymethylated chitosan nanoparticles on inhibiting the hepatocyte apoptosis after porcine hepatocyte xenotransplantation

作　　者：陈钟[1] 杨欣荣[1] 戴新征[1] 

机构地区：[1]南通大学附属医院肝胆外科,江苏省南通市226001

出　　处：《中国组织工程研究与临床康复》2007年第31期6178-6182,共5页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：江苏省自然科学基金(BK200544);江苏省卫生厅重大项目基金(H200215)~~

摘　　要：目的:观察聚乳酸-O-羧甲基壳聚糖纳米粒子对猪肝细胞异种移植后移植肝细胞凋亡的抑制作用。方法:实验于2004-03/2005-03在南通大学神经再生实验室完成,选用中国实验用小型猪(n=5)及SD大鼠(n=123)。①实验分组及方法:采用原位胶原酶循环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,按随机数字表法分成3组(n=41),分别在单纯肝细胞移植组、胶原肝细胞移植组、纳米胶原肝细胞移植组急性肝衰竭大鼠腹腔内移植震荡培养24h的猪肝细胞悬液、Ⅰ型胶原凝胶固定培养24h的猪肝细胞、Ⅰ型胶原凝胶包埋的聚乳酸-O-羧甲基壳聚糖纳米粒子培养24h的猪肝细胞。②实验评估:观察移植肝细胞的病理变化、培养和移植肝细胞的凋亡率、坏死率及活率。结果:①各组移植肝细胞的活率及凋亡率:纳米胶原肝细胞移植组移植肝细胞的存活时间最长。肝细胞体外培养1d后,3组肝细胞均存在不同程度的凋亡,以单纯肝细胞移植组凋亡率最高,纳米胶原肝细胞移植组最低。胶原肝细胞移植组、纳米胶原肝细胞移植组的移植肝细胞凋亡率随着移植时间的延长呈缓慢上升趋势,但较单纯肝细胞移植组为低;移植后1,2d,胶原肝细胞移植组、纳米胶原肝细胞移植组的移植肝细胞活率明显高于单纯肝细胞移植组(P<0.05);移植后3,5d时,纳米胶原肝细胞移植组移植肝细胞凋亡率明显低于胶原肝细胞移植组(P<0.05),活率则明显高于胶原肝细胞移植组(P<0.05)。②各组移植肝细胞的坏死率:移植后1d,胶原肝细胞移植组移植肝细胞坏死率明显上升;移植后2d,3组肝细胞坏死率均有不同程度的上升,胶原肝细胞移植组、纳米胶原肝细胞移植组明显高于单纯肝细胞移植组(P<0.05),胶原肝细胞移植组高于纳米胶原肝细胞移植组(P<0.05)。移植后3d,单纯肝细胞移植组肝细胞坏死率大幅度上升,胶原AIM： To evaluate the inhibition effect of polylactic acid （PLA）-O-carboxymethylated chitosan （CMC） nanoparticles on the apoptosis of the transplanted hepatocytes after porcine hepatocyte xenotransplantation. METHODS： The experiment was finished at Nerve Regeneration Laboratory of Nantong University from March 2004 to March 2005. Chinese miniature pigs （n =5） and SD rats （n =123） were used for experiment.①Recirculating collagenase perfusion method was used to isolate porcine hepatocytes, which then were cultured for 24 hours. Acute hepatic failure model in SD rats were established by the intraperitoneal injection of D-galactosamine. All the model rats were divided into 3 groups at random （n =41）, and were transplanted with free hepatocyte suspension （group A）, hepatocytes embedded in collagen gel （group B） and PLA-O-CMC nanoparticles-cultured hepatocytes embedded in collagen gel （group C） into peritoneal cavity, respectively. ②The pathological changes of the transplanted hepatocytes, the rate of apoptosis, viability and necrosis of the cultured and transplanted hepatocytes were observed. RESULTS： ①Viability and apoptosis of transplanted hepatocytes： The survival time of the transplanted hepatocytes was the longest in group C. The apoptosis of the cultured hepatocytes appeared in all groups on day 1 post-transplantation. The apoptosis rate was the highest in group A and the lowest in group C, It increased in lower level with the prolonged time of transplantation in groups B and C. The viability of the transplanted hepatocytes in groups B and C was obviously higher than that of group A on days 1 and 2 post-transplantation （P 〈 0.05）. The apoptosis rate of the transplanted hepatocytes in group C was significantly lower and the viability was significantly higher than that of group B on days 3 and 5 post-transplantation （P 〈 0.05）. ② The necrosis rate of the transplanted hepatocytes in group B increased significantly on day 1 post-transplantation. It incre

关 键 词：肝/细胞学 壳多糖 乳酸盐类 脱噬作用 肝功能衰竭 急性 细胞移植 

分 类 号：R318.08[医药卫生—生物医学工程]