摘要
目的探讨转染大鼠白细胞介素10（IL-10）基因的β2m^-／Thy-1^＋骨髓源性肝干细胞（BDLSC）移植对大鼠肝纤维化的治疗效果。方法分选Wistar大鼠的β2m^-／Thy-1^＋ BDLSC，转染大鼠IL-10基因。Wistar大鼠皮下注射CCl4，制作肝纤维化模型，分为3组：（1）模型组，经大鼠门静脉分支输注无菌生理盐水1ml；（2）BDLSC组，经大鼠门静脉分支输注β2m^-／Thy-1^＋ BDLSC悬液1ml（含2×10^5个细胞）；（3）IL-10组，经大鼠门静脉分支输注转染IL-10基因的β2m^-／Thy-1^＋ BDLSC悬液1ml（含2×10^5个细胞）；另以皮下注射橄榄油的Wistar大鼠为对照（正常组）。用细胞核染色剂二脒基苯基吲哚（DAPI）标记β2m^-／Thy-1^＋ BDLSC，以观察该细胞在肝内的定位情况。观察各组大鼠肝组织病理学改变和胶原的沉积情况，检测其肝功能和凝血功能。结果大鼠肝组织中可见DAPI标记的β2m^-／Thy-1^＋ BDLSC。模型组大鼠的肝组织病理学改变明显，BDLSC组病理学改变有所减轻，IL-10组的组织形态最接近正常组。模型组大鼠肝组织内胶原沉积明显增多，BDLSC组胶原沉积较少，IL-10组胶原沉积明显减少。IL-10组大鼠的血清丙氨酸转氨酶（ALT）、天冬氨酸转氨酶（AST）、碱性磷酸酶（ALP）和胆红素总量（TBIL）以及全血凝血酶原时间（PT）和活化部分凝血激酶时间（APTT）均低于模型组，差异均有统计学意义（P〈0．05），其中ALT、TBIL、PT和APTT均降至正常组水平，与BDLSC组相比较，差异有统计学意义（P〈0．05）。结论转染大鼠IL-10基因的β2m^-／Thy-1^＋ BDLSC移植对大鼠肝纤维化有一定的治疗效果。
Objective To explore the curative effectiveness of transplantation of bone marrow- derived liver stem cells （BDLSCs） transfected with rat interleukin-10 （IL-10） gene in the treatment of liver fibrosis in rats. Methodsβ2m^-/Thy-1^＋ BDLSCs isoJated from Wistar rats were transfected with adenovirus-mediated rat IL-10 gene. Wistar rats were subcutaneously injected with CCl4 to induce liver fibrosis, and randomly divided into three groups as follows; （1） the model group, infused with 1 ml normal saline （NS）; （2） the BDLSC group, infused with NS containing untreated β2m^-/Thy-1^＋ BDLSCs （2 ×10^5 cells）; （3） the IL-10 group, infused with NS containing β2m^-/Thy-1^＋ BDLSCs transfected with IL-10 gene （2 × 10^5 cells）. Infusion was done via the portal vein. Rats subcutaneously injected with olive oil served as control （the normal group）. The BDLSCs labeled with diamidine phenylindole dihydrochloride （DAPI） in the liver were localized. Pathological changes and collagen area in liver tissues were observed. Liver function and blood blotting function were tested. Results β2m^-/Thy-1^＋ BDLSCs labeled with DAPI were observed in liver tissues of rats. Significant pathological changes of liver tissues were observed in the model group. Compared with the model group, pathological changes were alleviated to some extent in the BDLSC group. The morphology of liver tissue in the IL-10 group was mostly close to that in the normal group. Collagen deposition of liver tissues was increased obviously in the model group. However, transplantations of untreated and IL-10-transfected BDLSCs both reduced collagen area. Compared with the BDLSC group, collagen deposition was significantly suppressed in the IL-10 group. Transplantation of IL-10-transfected BDLSCs suppressed obviously the levels of ALT, AST, ALP, TBIL, PT and APTT as compared with the model group （P〈0. 05）. The levels of ALT, TBIL, PT and APTT in the IL-10 group were significantly reduced to the normal levels as compared with those in the BDLSC group （P〈0. 05）. Conclusion Transplantation of BDI.SCs transfected with rat IL-10 gene was effective in treating liver fibrosis in rats. This combined strategy of IL-10 gene and BDLSCs may represent a feasible, effective and safe therapy form for liver fibrosis.
出处
《中华器官移植杂志》
CAS
CSCD
北大核心
2009年第7期403-406,共4页
Chinese Journal of Organ Transplantation
基金
国家自然科学基金（30500236）
