胎肝细胞输注移植联合运动对阻塞性黄疸大鼠肝损伤修复与再生的影响

目的:观察胎肝细胞输注移植联合中等强度有氧运动干预对大鼠阻塞性黄疸致重症肝损伤的修复与再生的影响。方法:40只SD大鼠随机分为假手术、阻塞性黄疸致肝损伤、胎肝细胞及胎肝细胞联合运动干预4组,后3组均采用改良的胆总管结扎法构建大鼠阻塞性黄疸致重症肝损伤模型。自孕龄12～16天的SD孕鼠胚胎中提取肝脏组织,加工制备胎肝细胞。两胎肝细胞组于模型构建过程中的胆总管结扎后进行胎肝细胞肝门静脉输注移植。运动组进行坡度5%、速度16.8 m/min、每天45 min、每周5天、共12周的中等强度跑台训练。12周后大鼠禁食过夜,随后分离出肝组织并取材,应用HE染色法观察大鼠肝细胞形态;生物化学法测定血清总胆红素(TBIL)、谷丙转氨酶(ALT)、谷草转氨酶(AST)含量;免疫组化技术检测肝组织表皮生长因子(EGF)、肝细胞生长因子(HGF)、多药耐药蛋白(MPR2)的蛋白表达;RT-PCR技术检测EGF、FXR、MRP2 mRNA表达。结果:假手术组肝细胞形态结构正常,阻塞性黄疸致肝损伤组可见肝细胞广泛空泡样变性、片状坏死,胎肝细胞组肝细胞少量点状坏死,胎肝细胞联合运动组肝细胞未见明显坏死。胎肝细胞组和胎肝细胞联合运动组大鼠血清TBIL、AST、ALT含量显著低于阻塞性黄疸致肝损伤组(P<0.05)。胎肝细胞组和胎肝细胞联合运动组大鼠HGF、EGF阳性表达显著低于阻塞性黄疸致肝损伤组(P<0.01),而MRP2阳性表达显著高于阻塞性黄疸致肝损伤组(P<0.05)。胎肝细胞组和胎肝细胞联合运动组大鼠EGF mRNA表达显著低于阻塞性黄疸致肝损伤组(P<0.01),而MRP2、FXR mRNA含量表达显著高于阻塞性黄疸致肝损伤组(P<0.05)。结论:胎肝细胞肝门静脉输注移植以及胎肝细胞肝门静脉输注移植联合中等强度有氧运动对阻塞性黄疸导致的重症肝损伤有明显的修复与再生作用,其机制可能是通过促进肝细胞再生相关因子的表达和肝细胞的增殖,从而发挥修复与再生作用。

The Effect of Fetal Liver Cell Infusion and Exercise on the Repairing and Regenerating Liver Injury in Rats with Obstructive Jaundice

Objective To observe the effect of moderate intensity aerobic exercise and fetal liver cell infusion on the repairing and regenerating liver injury in rat with obstructive jaundice.Methods Forty SD rats were divided into sham control(C),obstructive jaundice induced liver damage(OB),fetal liver cells(FL) and fetal liver cells plus exercise intervention(FL+E) groups.From the gestational 12 to 16 day-old SD rat embryos,fetal liver cells were prepared.FL group and FL+E group were given fetal liver cell through ...