转基因骨髓源性肝干细胞移植对大鼠肝纤维化的影响  

Effects of gene-transfected bone marrow-derived liver stem cells transplantation on liver fibrosis in rats

作　　者：宗春华[1] 孙超[1] 汪保灿[1] 陈源文[1] 陈颖伟[1] 孙巧玲[1] 李定国[1] 

机构地区：[1]上海交通大学医学院新华医院消化内科,上海200092

出　　处：《上海交通大学学报（医学版）》2007年第4期364-368,共5页Journal of Shanghai Jiao tong University:Medical Science

基　　金：国家自然科学基金(30500236);上海市教委基金(05BZ49);上海市科委基金(064119527)~~

摘　　要：目的探讨尿激酶型纤溶酶原激活物(uPA)基因修饰骨髓源性肝干细胞(BDLSC)移植对四氯化碳(CCl4)诱导的大鼠肝纤维化的影响。方法采用淤胆型肝损伤大鼠模型和含5%淤胆血清的培养液筛选和扩增BDLSC,使用RT-PCR方法检测培养2周时肝干细胞标志表达。体外将携带人uPA基因的腺病毒(AduPA)转染BDLSC并移植入CCl4诱导的肝纤维化大鼠体内,观察大鼠肝功能、胶原面积及病理组织学的变化。结果病理条件培养液培养2周的骨髓细胞形成肝细胞样集落,并表达胚胎期肝干细胞特征性蛋白。体内研究显示,与模型组和BDLSC组相比,BDLSC-uPA组大鼠血清丙氨酸转氨酶(ALT)、碱性磷酸酶(AKP)及总胆红素(TBIL)水平均有不同程度的降低,凝血酶原时间(PT)时间缩短,白蛋白(ALB)水平升高,肝组织细胞外基质(ECM)水平及羟脯氨酸(Hyp)含量明显降低;肝组织结缔组织增生程度减轻,假小叶形成不明显,胶原面积明显减少。结论uPA基因修饰BDLSC移植能有效抑制CCl4诱导的大鼠肝纤维化,改善纤维化大鼠的肝功能,从而为转基因BDLSC移植治疗肝纤维化的临床应用提供理论依据和实践基础。Objective To explore the effects of urokinase type plasminogen activator（uPA） gene-modified bone marrow-derived stem cells （BDLSCs） transplantation on CCl4- induced liver fibrosis in rats. Methods BDLSCs were selected and amplified by setting up cholestatic hepatic injured model and preparing a pathological conditioned medium containing 5% cholestatic serum. RT-PCR method was applied to determine the expression of characteristics of liver stem cells. BDLSCs were transfected by AduPA in vitro and transplanted into rats with CCl4-induced liver fibrosis. Liver functions, areas of collagen and pathohistology were observed. Results Bone marrow cells were able to form hepatocyte-like colonies in the presence of pathological conditioned medium, and express characteristical proteins of fetal hepatocytes. Studies in vivo showed that in the BDLSC-uPA group the serum levels of alanine aminotransferase （ALT） , alkaline phosphatase （AKP） and total bilirubin （TBIL） decreased dramatically, prothrombin time （PT） shortened, the serum albumin level （ALB） was higher, and serum levels of extracellular matrix （ECM） and hydroxyproline （Hyp） content of the liver were much lower, the extent of the fibrotic tissues of the liver was much less, and the formation of pseudolobules were not obvious, the areas of collagen were markedly smaller, compared with those in the control group of rats infused by normal saline and BDLSCs. Conclusion uPA gene-modified BDLSCs transplantation may suppress hepatic fibrosis and ameliorate liver functions in fibrotic rats induced by CCIa injection, which provides theoretical basis and practical foundation for the clinical application of gene-modified BDLSCs in treating liver fibrosis.

关 键 词：尿激酶型纤溶酶原激活物 骨髓源性肝干细胞 肝纤维化 

分 类 号：R575.2[医药卫生—消化系统] R-332[医药卫生—内科学]