输注转染人IL-10基因的骨髓间充质干细胞延长异种肝移植大鼠存活时间

Perfusion of MSCs transfected by human IL-10 gene prolonging survival of rat liver xenografts

目的探讨人白细胞介素10（hIL-10）修饰的豚鼠骨髓间充质干细胞（hIL-10-MSCs）输注对异种肝移植大鼠存活的影响及其可能机制。方法以携带hIL-10基因的慢病毒（hIL-10／LOX-cwGFP）为载体构建hIL-10-MSCs。以豚鼠为供者，Wistar大鼠为受者，制作非协调性异种原位肝移植模型，将受者随机分成3组，每组60只：空白对照组大鼠术前3天和术后1天各静脉注射生理盐水和地塞米松1次；MSCs组大鼠术前3天和术后1天各静脉注射MSCs和地塞米松1次；hIL-10-MSCs组大鼠术前3天和术后1天各静脉注射hIL-10-MSCs和地塞米松1次。记录供者手术时间、供肝冷缺血时间、受者手术时间、肝上下腔静脉（SVC）吻合时间、无肝期及受者存活时间，测定术后各组受者血清丙氨酸转氨酶（ALT）、天冬氨酸转氨酶（AST）和乳酸脱氢酶（LDH）的含量，酶联免疫吸附试验法测定各组受者移植肝组织中自细胞介素2（IL-2）、IL-4、hIL-10、IL-12、IL-15、IL-18和TGF目的含量，逆转录聚合酶链反应法检测各组受者移植肝组织中上述各细胞因子mRNA的表达。结果hIL-10-MSCs组受者存活时间为（72．0±5．5）h，空白对照组和MSCs组受者的存活时间分别为（29．0±2．2）h和（48．0±4．6）h，前者明显长于后二者，差异有统计学意义（P〈0．05，P〈0．05）。hIL-10-MSCs组移植肝组织中IL-4和TGF-β。mRNA的表达明显高于空白对照组和MSCs组（P〈0．05，P〈0．05），而IL-2、IL-12、IL-15、IL-18mRNA的表达明显低于空白对照组和MSCs组（P〈0．05，P〈0．05），hIL-10-MSCs组肝组织中hIL-10mRNA的表达明显高于空白对照组和MSCs组。结论静脉输注hIL-10-MSCs可延长异种肝移植大鼠的存活时间，其机制可能与hIL-10-MSCs抑制IL-2、IL-12、IL-15、IL-18等细胞因子的表达，促进IL-4和TGF-β1的表达有关。

Objective To observe the influence of hlL-10 modified MSCs from guinea pigs on survival of rat discordant liver xenografts and the possible mechanisms. Methods hIL-10-MSCs were constructed by transfection with lentivirus hIL-10/LOX-cwGFP. Model of guinea pig to rat discordant liver xenotransplantation was set up. The recipients were divided into 3 groups （each group having 60 rats） ： the control group, injected with NS and dexamethasone; the MSCs group, injected with MSCs and dexamethasone; the hIL-10 MSCs group, injected with hIL-10-MSCs and dexamethasone. Operating time of donors and recipients, cold ischemia time of donor liver, anastomosis time of SVC, anhepatie time, and survival time of recipients were recorded. The contents of ALT, AST, and LDH after operation were determined. The levels of IL-2, IL4, hIL-10, IL-12, IL-15, IL-18 and TGF-β1 mRNA and protein in the liver were detected by RT-PCR and ELISA, respectively. Results Survival time of hIL-10-MSCs, control and MSCs groups was （72. 0 ± 5.5）, （29. 0 ± 2. 2） and （48. 0 ± 4. 6） h, respectively. The survival time in hIL-10-MSCs group was significantly longer than in control and MSCs groups （both P〈0. 05）. The expression levels of IL-4 and TGF-β1 mRNA were significantly increased in hIL-10-MSCs group as compared with control and MSCs groups （both P（0. 05）, but those of IL-2, IL-12, IL-15, and IL-18 mRNA were significantly reduced （both P〈0.05）. hIL-10 mRNA expression level in the liver tissues in hIL-10-MSCs group were significantly higher than in control and MSCs groups. Conclusion Intravenous perfusion of hIL-10 MSCs can prolong survival of rat liver xenografts by decreasing the expression of IL-2, IL-12, IL-15 and IL 18, and increasing the expression of IL-4 and TGF-β1.