CAJ | 학술논문

背景：近来国内外研究证明，来自其他组织的干细胞能够归巢到肝脏，并可能参与肝组织的再生，这为发展干细胞治疗肝脏疾病提供了新的希望。目的：探究人脐带源间充质干细胞的分离、培养方法，观察脐带间充质干细胞移植对大鼠肝纤维化模型的修复作用，为脐带间充质干细胞的临床应用提供可靠的理论依据。方法：自然贴壁法分离、纯化人脐带间充质干细胞并进行体外培养和扩增，用皮下多点注射CCl4制备肝纤维化大鼠模型。将22只模型大鼠随机分为模型损伤组(n=11)和细胞移植组(n=11)。细胞移植组在模型制备成功后的第1，2，3周经尾静脉给予1×106脐带间充质干细胞治疗，4周后将大鼠处死，收集各组大鼠血液检测肝功能；摘取肝脏行苏木精-伊红染色，观察病理变化；免疫组化法观察库普弗细胞的数量及分布；免疫组化法观察治疗组脐带间充质干细胞的定位情况。结果与结论：脐带间充质干细胞经尾静脉移植入肝硬化大鼠后，大鼠的肝功能均明显改善，与对照组比较，差异有显著性意义(P<0.05)；肝组织苏木精-伊红染色提示，肝纤维化程度明显改善；免疫组化法观察库普弗细胞的数量提示，库普弗细胞数量明显减少；免疫组化方法利用抗BrdU抗体在治疗组大鼠肝脏观察到BrdU标记的脐带间充质干细胞。说明脐带间充质干细胞移植可以改善大鼠外周血液的血生化特性和肝的组织学结构。
배경：근래국내외연구증명，래자기타조직적간세포능구귀소도간장，병가능삼여간조직적재생，저위발전간세포치료간장질병제공료신적희망。목적：탐구인제대원간충질간세포적분리、배양방법，관찰제대간충질간세포이식대대서간섬유화모형적수복작용，위제대간충질간세포적림상응용제공가고적이론의거。방법：자연첩벽법분리、순화인제대간충질간세포병진행체외배양화확증，용피하다점주사CCl4제비간섬유화대서모형。장22지모형대서수궤분위모형손상조(n=11)화세포이식조(n=11)。세포이식조재모형제비성공후적제1，2，3주경미정맥급여1×106제대간충질간세포치료，4주후장대서처사，수집각조대서혈액검측간공능；적취간장행소목정-이홍염색，관찰병리변화；면역조화법관찰고보불세포적수량급분포；면역조화법관찰치료조제대간충질간세포적정위정황。결과여결론：제대간충질간세포경미정맥이식입간경화대서후，대서적간공능균명현개선，여대조조비교，차이유현저성의의(P<0.05)；간조직소목정-이홍염색제시，간섬유화정도명현개선；면역조화법관찰고보불세포적수량제시，고보불세포수량명현감소；면역조화방법이용항BrdU항체재치료조대서간장관찰도BrdU표기적제대간충질간세포。설명제대간충질간세포이식가이개선대서외주혈액적혈생화특성화간적조직학결구。
BACKGROUND:Recent studies verified that other tissues-derived stem cells can be homing to the liver, possibly participate in the regeneration of liver tissues, which provides new hope for stem cells in treatment of liver disease. OBJECTIVE:To observe isolation and culture of human umbilical cord-derived mesenchymal stem cells, to observe the repairing effect of umbilical cord-derived mesenchymal stem celltransplantation on hepatic fibrosis, and then to provide a reliable theoretical basis for further clinical application of umbilical cord-derived mesenchymal stem cells. METHODS:Human umbilical cord-derived mesenchymal stem cells were isolated and purified by natural adherent method, and then cultured and amplified in vitro. A rat model of hepatic fibrosis was prepared using subcutaneous multi-point injection of CCl4. A total of 22 rat models were randomly assigned to model injury group (n=11) and celltransplantation group (n=11). At 1, 2 and 3 weeks after model induction, the rats in the celltransplantation group were treated with 1×106 umbilical cord-derived mesenchymal stem cells via caudal vein. Four weeks later, the rats were sacrificed. Blood of rats was col ected from each group to detect liver functions. The liver was removed to receive hematoxylin-eosin staining, and pathological changes were observed. The number and distribution of Kupffer’s cells were observed using immunohistochemistry. The localization of umbilical cord-derived mesenchymal stem cells from treatment group was observed using immunohistochemistry.RESULTS AND CONCLUSION:After umbilical cord-derived mesenchymal stem cells were infused into rats with cirrhosis via caudal vein, liver function was significantly improved, which showed significant differences as compared with the control group (P<0.05). Hematoxylin-eosin staining revealed that hepatic fibrosis was apparently improved. Immunohistochemistry results demonstrated that the number of Kupffer’s cells was obviously reduced, and BrdU-labeled umbilical cord-derived mesenchymal stem cells were visible in rat liver of the treatment group using anti-BrdU antibody. These results suggested that umbilical cord-derived mesenchymal stem celltransplantation could improve biochemical characteristics of rat peripheral blood and histological structure of liver.