人羊水干细胞移植治疗肝硬化大鼠的实验研究

Therapeutic effect of transplanting human amniotic fluid stem cells on rat cirrhosis

目的了解人羊水干细胞(hAFSC)移植对四氯化碳诱导肝硬化大鼠的治疗作用。方法使用贴壁法分离培养羊水干细胞,Western blot鉴定。Wistar大鼠24只随机分为对照组8只和肝硬化组16只,肝硬化组皮下注射四氯化碳植物油,7周后再随机分为肝硬化模型对照组(注射等体积PBS,n=8)、hAFSC移植组(n=8)。直接经肝内注射hAFSC,3周后处死所有大鼠,收取血样进行生化检查,肝组织石蜡切片进行病理学分析。结果分离的羊水干细胞均表达特异性标记物Oct-4、SSEA-4。与大鼠肝硬化模型组比较,hAFSC移植组血清Alb明显升高,ALT、AST显著降低。肝硬化程度明显减轻,假小叶附近脂肪化细胞减少,肝组织中胶原纤维的量明显减少。结论 hAFSC肝内移植可减轻大鼠肝硬化病变程度,改善肝功能。

Objective To study the effect of human arnniotic fluid stem cell （hAF5C） transplantation on rat liver cirrhosis induced by carbon tetrachloride. Methods hAFSC were isolated and cultured with adherent method and identified by Western blot. Experimental Wister rat （n = 24） were randomly divided into control group（n = 8） and liver cirrhosis group （n = 16）, which were induced by injection of 60% carbon tetraehloride and plant oil complex for 7 weeks. After 3 weeks rats were challenged with CCL4, liver cirrhosis group were randomly divided into model group （ only infusing saline, n = 8 ）, hAFSC transplantation group （n = 8）, which hAFSC was directly intrahepatic injected. All rats were sacrificed 3 weeks after transplantation. Blood biochemical examination was performed, liver tissue paraffin was histological analyzed. Results Serum albumin was significantly elevated in isolated hAFSC transplantation group, serum alanine aminotransferase （ALT）and aspartate aminotransferase （AST）were reduced, the extent of liver cirrhosis was significantly ameliorated, the numbers of fat cells surrounding pseudolobuli and liver tissue collagen fibers were markedly decreased, respectively. Conclusion hAFSC transplantation could ameliorate liver cirrhosis, and improve the function of liver.