输注转染人IL-10基因的骨髓间充质干细胞减轻大鼠异种移植肝脏的细胞凋亡

The role of hIL-10-MSCs in apoptosis of rat discordant liver xenotransplantation

目的 探讨转染人白细胞介素10（Hil-10）基因的骨髓间充质干细胞（MSC）对大鼠异种移植肝脏细胞凋亡的影响.方法 以携带Hil-10基因的慢病毒Hil-10/LOX-cwGFP转染豚鼠的MSC.以豚鼠为供者,Wistar大鼠为受者,利用两套管法行原位肝移植.空白对照组的受者分别于术前24 h和术后24 h经股静脉注射生理盐水2 ml＋地塞米松0.5 ml（2 mg/ml）各1次;MSC组的受者分别于上述时间点自股静脉注射2.0×106/ml的MSC单细胞悬液2 ml＋地塞米松0.5 ml（2mg/ml）各1次;Hil-10-MSC组的受者分别于上述时间点自股静脉注射2.0×106/ml的Hil-10-MSC单细胞悬液2 ml＋地塞米松0.5 ml（2 mg/ml）各1次.术后12 h切取移植肝脏,观察移植肝组织学变化,测定肝组织中Hil-10、凋亡蛋白酶-3（Caspase-3）、Fas和FasL的表达,观察肝细胞的凋亡情况.结果 与空白对照组、MSC组相比,Hil-10-MSC组的病理改变最轻,IL-110表达明显增加,FasL明显减少.Hil-10-MSC组的Fas和Caspase-3的阳性区域面积分别为11.5%和25.1%,明显低于空白对照组的35.3%和70.8%（P＜0.05）.Hil-10-MSC组的凋亡指数为32.5%,明显低于空白对照组的74.1%和MSC组的50.3%（P＜0.05）.结论 Hil-10-MSC可明显减轻大鼠异种移植肝脏的细胞凋亡,其机制可能与其抑制Fas/FasL的表达有关.

Objective To observe the influence of hIL-10-MSCs on apoptosis of rats subject to discordant liver xenotransplantation. Methods The model of guinea pig to rat discordant liver xenotransplantation was set up. The orthotopic liver transplantation model was established by using modified two-cuff technique. Following groups were designed： control group, MSCs group and hIL-10MSCs group. Before and after operation, the recipients in control group were injected with normal sodium （2 ml） and decaesadril （0.5 ml） ;Those in MSCs group were injected with MSCs （4.0× 106/ml）and decaesadril （0.5 ml）; Those in hIL-10-MSCs group were injected with hIL-10-MSCs （4.0 × 106/ ml）and decaesadril （0.5 ml）. Twelve h after operation, the livers of recipient were observed by HE staining and electron microscope, and apoptosis of the livers was detected by using TUNEL. The expression levels of hIL-10, caspase-3, Fas and FasL were assayed by Western blot or immunohistochemistry. Results As compared with control group and MSCs group, the pathological changes was alleviated, the expression of IL-10 was significantly increased, the expression of FasL was significantly reduced in hIL-10-MSCs group. The positive area size of Fas and Caspase-3 in hIL-10MSCs group was 11.5 % and 25.1 %, respectively, significantly smaller than those in control group （35.3 % and 70.8 % respectively, P＜0.05. Apoptosis index in hIL-10-MSCs group was 32.5 %,which was significantly lower than in control group （74.1%） and MSCs group （50. 3 % ）. ConclusionhIL-10-MSCs can notably decrease apoptosis of the transplanted liver probably by inhibiting theexpression of Fas/FasL.