摘要
背景:课题组前期实验用I型胶原包埋聚乳酸-O-羧甲基壳聚糖纳米粒子黏附培养的猪肝细胞治疗急性肝衰竭大鼠取得了良好的疗效。目的:进一步观察聚乳酸-O-羧甲基壳聚糖纳米粒子在猪肝细胞腹腔内移植治疗急性肝衰竭大鼠过程中对免疫排斥反应的影响。设计、时间及地点:随机对照动物实验,于2005-05/2006-05在南通大学肝胆外科研究所和神经再生实验室完成。材料:原位胶原酶循环肝灌注法分离猪肝细胞;SD大鼠64只D-氨基半乳糖腹腔内注射制作急性肝衰竭模型。方法:64只模型大鼠随机分为4组:模型组不干预;纳米胶原肝细胞组将Ⅰ型胶原凝胶包埋的聚乳酸-O-羧甲基壳聚糖纳米粒子黏附培养24h的猪肝细胞植入大鼠腹腔内,并用大网膜适当包裹;胶原肝细胞组将Ⅰ型胶原凝胶固定培养24h的猪肝细胞植入大鼠腹腔内;单纯肝细胞组将振荡培养24h的猪肝细胞悬液直视下注入大鼠的小网膜囊内。移植在造模48h后进行,移植细胞量均为5.0×107个肝细胞。主要观察指标:移植后1,2,3,5,7d取大鼠血清检测白细胞介素2、干扰素γ、IgG、IgM浓度,以模型组数据为移植前指标;移植后1,3,7d取腹腔移植物光镜下观察移植肝细胞的病理变化。结果:移植后7d内各组血清白细胞介素2、干扰素γ水平差异无显著性。各组血清IgG水平在移植后逐渐升高,第3天达到最高峰,随后逐渐下降;移植后2,3d,纳米胶原肝细胞组血清IgG质量浓度低于其他2组(P<0.05),至移植后7d,纳米胶原肝细胞组血清IgG水平高于单纯肝细胞组(P<0.05)。各组血清IgM质量浓度在移植后第1,2天明显低于移植前(P<0.05),移植后第3～7天逐渐升高,其中纳米胶原肝细胞组IgM质量浓度高于其他2组(P<0.05)。移植后第3天胶原肝细胞组移植肝细胞数量明显减少,单纯肝细胞组则几乎找不到存活的肝细胞,而纳米胶原肝细胞组移植后第7天仍能找到存活肝细胞。结论:聚乳酸-O-羧甲基壳聚糖纳米粒子在猪肝细胞腹腔内移植治疗大鼠急性肝衰竭过程中对体液免疫排斥反应具有一定的抑制作用。
BACKGROUND： Previous research has proved good therapeutic effect on the acute hepatic failure by the treatment of polylactic acid-O-carboxymethylated chitosan （PLA-O-CMC） nanoparticles-cultured porcine hepatocytes embedded in type Ⅰ collagen.
OBJECTIVE： To investigate the effect of PLA-O-CMC nanoparticles on immunological rejection in intraperitoneal porcine bepatocyte xenotransplantation for treatment of acute liver failure rats. DESIGN,TIME AND SETTING： Randomized control animal experiments were carried out at the Institute of Hepatobiliary Surgery and Laboratory of Nerve Regeneration in Nantong University from May 2005 to May 2006.
MATERIALS： In situ recirculating collagenase perfusion method was used to isolate porcine hepatocytes. 64 SD rats were intraperitoneally injected with D-galactosamine to induce acute hepatic failure models.
METHODS：Totally 64 rats were divided into four groups： Model group received no intervention; Nano-collagen hepatocytes group was transplanted with PLA-O-CMC nanoparcicles-cultured porcine hepatocytes embedded in collagen gel, which was then wrapped up using greater omentum; Porcine hepatocytes embedded in collagen gel （collagen hepatocytes group） and free porcine hepatocytes suspension （pure hepatocytes group） were transplanted into peritoneal cavity of SD rats, respectively. At 48 hours following the modeling, the porcine hepatocytes transplantations were performed and the number of transplanted cells was 5.0× 10^7 hepatocytes in all groups.
MAIN OUTCOME MIEASURES： The serum interleukin-2, interferon- γ, IgG and IgM levels of rats were determined at days 1, 2, 3, 5 and 7. The pathological changes of transplants were observed under microscope at days 1, 3 and 7.
RESULTS： There were no significant differences in serum interleukin-2 and interferony levels of all groups during 7 days post-transplantation. Serum IgG levels increased with a peak at day 3 after hepatocytes transplantation, and then began to decline. Serum IgG levels in nano-collagen hepatecytes group were lower than those in collagen hepatocytes group and pure hepatocytes group at days 2 and 3 （P 〈 0.05）. Serum IgG levels in nano-collagen hepatocytes group were higher than those in pure hepatocytes group at day 7 （P 〈 0.05）. At days 1 and 2, serum IgM levels in three groups were lower than those pre-transplantation （P 〈 0.05）. From days 3 to 7. serum IgM levels increased with time in three groups, and the highest level was found in nano-collagen l hepatocytes group （P 〈 0.05）. At day 3, the number of transplanted hepatocytes in collagen hepatocytes group significantly Ⅰ decreased, and transplanted hepatocytes almost could not be found in pure hepatocytes group. On day 7, they could be found in nano-collagen hepatocytes group.
CONCLUSION： PLA-O-CMC nanoparticles can inhibit the immunological rejection in intraperitoneal porcine hepatocyte xenotransplantation for treatment of acute liver failure rats.
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2008年第32期6287-6291,共5页
Journal of Clinical Rehabilitative Tissue Engineering Research
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)~~
