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）诱导肝纤维化大鼠肝细胞再生的影响。方法纯系Fisher344雄性大鼠10只，为BDLSC供体大鼠。体外将AduPA转染雄性大鼠BDLSC。纯系Fisher344雌性大鼠36只，均分为正常组（皮下注射橄榄油）、模型组（CCI4造模，尾静脉注射0．9％氯化钠）、BDLSC组（CCl4造模，尾静脉输入BDLSC）、转基因组（CCl4造模，尾静脉输入转基因BDLSC）。检测大鼠肝功能和肝组织胶原面积。半定量RT—PCR方法检测大鼠肝组织肝细胞生长因子（HGF）及其受体c—metmRNA表达水平。免疫组织化学法检测大鼠肝组织增殖细胞核抗原（PCNA）蛋白表达。结果正常组、模型组、BDLSC组、转基因组肝组织胶原染色面积分别为0．12％±0．03％、14，49％±1．40％、8．25％±0．82％、5．12％±0．40％，组间差异均有统计学意义（P值均〈0．05）。与模型组和BDLSC组相比，转基因组大鼠肝功能明显改善，血清透明质酸（HA）、血清Ⅲ型前胶原（PCUI）、肝组织羟脯氨酸含量明显降低，肝组织HGF和c—metmRNA水平均显著上调，PCNA蛋白表达显著增加。结论uPA基因修饰BDLSC移植可能诱导肝细胞增殖，从而改善CCl4诱导肝纤维化大鼠的肝功能。
Objective To explore the effects of urokinase type plasminogen activator （uPA） gene-modified bone marrow derived liver stem cells （BDLSC） transplantation on hepatocyte regeneration in CCl4 induced liver fibrosis rats. Methods Ten male Fisher 344 rats were donor rats of BDLSC. The BDLSC of male rat was transfected with AduPA. Thirty-six female Fisfier 344 rats were equally divided into normal group （injected subcutaneously with olive oil）, model group （CCL induced the model, injected through tail vein with 0.9% sodium chloride）, BDLSC group （CCl4 induced the model, injected through tail vein with BDLSC） and gene transfected group （CCl4 induced the model, injected through tail vein with gene transfected BDLSC）. Liver function and area of collagen were observed. The expression of hepatic growth factor （HGF） and its receptor c-met mRNA in rats＇ liver tissues were tested by semiquantitative RT-PCR. The expressions of proliferating cell nuclear antigen （PCNA） in rats＇ liver tissues were determined by immunohistochemistry staining. Results The areas of collagen in normal group, model group, BDLSC group and gene transfected group was 0. 12% ± 0.03%, 14.49%±1.40%, 8.25% ±0.82% and 5.12%±0.40% accordingly, there were significant differences between groups （P〈0.05）. Compared with model group and BDLSC group, the liver function of gene transfected group significantly improved, the serum levels of hyaluronic acid （HA）, procollagen Ⅲ （PCⅢ） and the content of hydroxyproline in liver tissues decreased dramatically. The expression of HGF and c-met at mRNA levels were up-regulated significantly, and the expression of PCNA protein in liver tissues increased obviously. Conclusion uPA gene-modified BDLSC transplantation may induce proliferation of hepatocytes, and then improve the liver functions of fibrotic rats induced by CCl4.
出处
《中华消化杂志》
CAS
CSCD
北大核心
2011年第5期307-311,共5页
Chinese Journal of Digestion
基金
国家自然科学基金资助项目（30500236）
上海市教委科研资助项目（05BZ49）
上海市科委医学临床研究重点科技攻关资助项目（064119527）
上海市教委优秀青年教师科研专项基金资助项目（jdy06040）
关键词
尿纤溶酶原激活物
干细胞移植
肝细胞
肝硬化
肝细胞生长因子
增殖细胞核
抗原
四氯化碳
疾病模型
动物
Uninary plasminogen activator
Stern cell transplantation
Hepatocytes
Liver cirrhosis
Hepatocyte growth factor
Proliferating cell nuclear antigen
Carbon tetrachloride
Disease models, animal
