载肝细胞生长因子的纳米粒子治疗急性肝衰竭大鼠

Therapeutic effects of hepatocyte transplantation using hepatocyte growth factor-loaded polylactic acid-O-carboxymethylated chitosan nanoparticles on treating rats with acute liver failure

目的 观察载肝细胞生长因子（HGF）的聚乳酸-O-羧甲基壳聚糖（PLA-O-CMC）纳米粒子培养的大鼠肝细胞腹腔内移植对急性肝衰竭（ALF）大鼠的治疗效果.方法 分别将5 ml培养24 h的大鼠肝细胞（5.0×107个）（Ⅰ组）、PLA-O-CMC纳米粒子培养的大鼠肝细胞（Ⅱ组）、载HGF的PLA-O-CMC纳米粒子培养的大鼠肝细胞（Ⅲ组）移植到ALF大鼠腹腔内,以PLA-O-CMC纳米粒子培养的大鼠肝细胞腹腔移植加每天静脉注射HGF 10 μg/kg×7 d（Ⅳ组）和5 ml RPMI 1640培养基腹腔内注射（Ⅴ组）作为对照.观察受体大鼠存活率、肝功能、肝组织光镜和电镜变化.结果 移植后14d大鼠的存活率Ⅰ～Ⅴ组分别为50.00％、68.75％、81.25％、75.00％和18.75％,各移植组高于对照组,Ⅲ组最高.移植后24 h,Ⅱ～Ⅳ组各项肝功能指标开始好转,至移植后7d,各组间除谷丙转氨酶（ALT）外,差异无统计学意义（P＞0.05）.Ⅲ组的肝功能和肝脏病理损害恢复最快,其次为Ⅳ、Ⅱ、Ⅰ组,Ⅴ组恢复最慢、最差.结论 应用载HGF的PLA-O-CMC纳米粒子培养的大鼠肝细胞腹腔内移植治疗ALF大鼠能逆转肝功能,提高生存率,较静脉途径给予HGF的治疗效果更好.

Objective To evaluate the therapeutic effects of intraperitoneal hepatocyte transplantation （HCT） of hepatocyte growth factor （HGF）-loaded polylactic acid-O-carboxymethylated chitosan （PLA-O-CMC） nanoparticles on acute liver failure （ALF） rats.Methods Five ml rat hepatocyte suspension cultured for 24 h （5.0 × 107 cells） （group Ⅰ）,rat hepatocytes co-cultured with PLA-O-CMC nanoparticles （group Ⅱ） and rat hepatocytes co-cultured with HGF-loaded PLA-O-CMC nanoparticles （group Ⅲ） were transplanted into the abdominal cavity of ALF rats respectively.Rat hepatocytes co-cultured with PLA-O-CMC nanoparticles were transplanted into the abdominal cavity of ALF rats,and HGF was intravenously injected （10 μg/kg ×7 day） （group Ⅳ）,or 5 ml RPMI 1640 was injected （group Ⅴ）.The 14-day survival rate,hepatic function and the pathological changes of the liver under light and electron microscope in ALF rats were observed.Results There was significant difference in the 14-day survival rate among group Ⅰ （50.00％）,group Ⅱ （68.75％）,group Ⅲ （81.25％）,group Ⅳ （75.00％） and group Ⅴ （18.75％） （P ＜0.05）.The survival rate in group Ⅲ was the highest in the five groups.Twenty-four h after HCT,the improvement of hepatic functional parameters showed significant difference between groups Ⅱ,Ⅲ,Ⅳ with group Ⅴ.At 7th day after HCT,there was no significant defference among all groups except alanine aminotransferase （ALT）.The hepatic function and pathological damage were quickly restored in group Ⅲ,follwed by group Ⅳ,group Ⅱ and group Ⅰ,with the slowest in group Ⅴ.Conclusion Intraperitoneal transplantation of HGF-loaded PLA-O-CMC nanoparticle-attached hepatocytes can increased the survival rate of ALF rats,can reverse hepatic function,and promote regeneration of the damaged recipient liver.