CAJ | 학술논문

目的：探讨脐带间充质干细胞（hUCMSC）移植对急性肝功能衰竭大鼠的治疗作用，并优化移植治疗途径。方法采用组织块贴壁法收集 hUCMSC，流式细胞术检测细胞表面标志物。将 SD 大鼠随机分为经尾静脉注射移植治疗组，经肝叶注射移植治疗组，模型对照组，空白对照组。急性肝衰竭动物模型采用一次性腹腔注射2．5 mL／kg CCl4橄榄油（1∶1）混合溶液建立，24 h 后移植治疗组分别经尾静脉和肝叶注射 hUCMSC 细胞悬液；治疗后0 h、24 h、48 h、72 h、96 h和1周收集血清分析肝功能恢复情况；治疗后3 d、1周、2周留取肝脏标本，观察病理学恢复情况，用 Real-time PCR 检测鼠肝中人 CK8、CK18和 AFP 基因 mRNA 转录水平，免疫组化法检测人 CK18的表达。结果移植治疗组 TBil 和 ALT 含量较模型对照组有明显恢复（P ＜0．05），肝细胞再生程度提高，炎细胞减少，肝脏病理学修复作用增强。在造模后1周、2周时，经尾静脉移植治疗组和经肝叶注射移植治疗组的肝组织中，CK8、CK18和 AFP 基因 mRNA 的相对转录量与模型对照组相比差异有统计学意义（P ＜0．05）。经尾静脉注射移植治疗组和经肝叶移植治疗组中，移植后的 hUCMSC 在鼠肝中诱导分化后表达人肝细胞相关蛋白 CK18。经尾静脉和经肝叶注射移植治疗组的肝功能、移植细胞分化程度比较差异无统计学意义（P ＞0．05）。结论采用 hUCMSC 移植治疗急性肝衰竭大鼠模型后，能促进肝功能和肝脏病理学修复。移植后的 hUCMSC 自身能分化为具有肝细胞功能的类肝样细胞。经尾静脉与经肝叶注射移植疗效相似。
목적：탐토제대간충질간세포（hUCMSC）이식대급성간공능쇠갈대서적치료작용，병우화이식치료도경。방법채용조직괴첩벽법수집 hUCMSC，류식세포술검측세포표면표지물。장 SD 대서수궤분위경미정맥주사이식치료조，경간협주사이식치료조，모형대조조，공백대조조。급성간쇠갈동물모형채용일차성복강주사2．5 mL／kg CCl4감람유（1∶1）혼합용액건립，24 h 후이식치료조분별경미정맥화간협주사 hUCMSC 세포현액；치료후0 h、24 h、48 h、72 h、96 h화1주수집혈청분석간공능회복정황；치료후3 d、1주、2주류취간장표본，관찰병이학회복정황，용 Real-time PCR 검측서간중인 CK8、CK18화 AFP 기인 mRNA 전록수평，면역조화법검측인 CK18적표체。결과이식치료조 TBil 화 ALT 함량교모형대조조유명현회복（P ＜0．05），간세포재생정도제고，염세포감소，간장병이학수복작용증강。재조모후1주、2주시，경미정맥이식치료조화경간협주사이식치료조적간조직중，CK8、CK18화 AFP 기인 mRNA 적상대전록량여모형대조조상비차이유통계학의의（P ＜0．05）。경미정맥주사이식치료조화경간협이식치료조중，이식후적 hUCMSC 재서간중유도분화후표체인간세포상관단백 CK18。경미정맥화경간협주사이식치료조적간공능、이식세포분화정도비교차이무통계학의의（P ＞0．05）。결론채용 hUCMSC 이식치료급성간쇠갈대서모형후，능촉진간공능화간장병이학수복。이식후적 hUCMSC 자신능분화위구유간세포공능적류간양세포。경미정맥여경간협주사이식료효상사。
Objective To explore treatment effect and optimize the approach of human umbilical cord mesenchymal stem cells (hUCMSCs)transplantation on acute hepatic failure in rats.Methods hUCMSCs were isolated from umbilical cord with tissue explants adherent method,and the surface antigens of which were tested by flow cytometry.Forty-eight male Sprague Dawley rats were randomly divided into four groups,including transplantation treatment via tail vein group, transplantation treatment via hepatic lobe group,liver failure animal model group,and control group.Animal model of acute liver failure was induced by injecting intraperitoneally with 50% olive oil solution of carbon tetrachloride 2.5 ml·kg-1 ,then treatment groups were injected with hUCMSCs suspension separately through tail vein or injection directly into the liverhepatic lobe after 24h post-animal model built 24h.Blood serum and liver tissues were collected in several times to analyze liver function improvement and histological repair.Real-time PCR was used to detect the expression of human CK8, CK18 and AFP mRNA in liver tissues.Immunohistochemistry was used to detect the expression of human CK18 in liver tissues.Results There were statistically significant differences among liver function indexes after transplantation (P <0.05).hUCMSCs improved histological status through enhancing hepatocellular regeneration and reducing inflammatory cells.Real-time PCR results showed that the expression of CK8,CK18 and AFP mRNA were obviously increased in the tail vein transplantation group and hepatic lobe injection transplantation group,when compared with the model group (P <0.05).Immunochemistry results revealed that transplanted hUCMSCs in animal liver could differentiate into functional hepatocyte-like cells that expressed human CK18 as hepatocyte-specific marker in both tail vein transplantation group and hepatic lobe injection transplantation group.No significant differences of histological repair and grade of differentiation were examined between the treatments through tail vein group or direct injection into hepatic lobe group (P >0.05).Conclusion There were statistically significant differences among liver function indexes after transplantation (P <0.05 ).hUCMSCs improved histological status through enhancing hepatocellular regeneration and reducing inflammatory cells.Real-time PCR results showed that the expression of CK8,CK18 and AFP mRNA were obviously increased in the tail vein transplantation group and hepatic lobe injection transplantation group,when compared with the model group (P <0.05).Immunochemistry results revealed that transplanted hUCMSCs in animal liver could differentiate into functional hepatocyte-like cells that expressed human CK18 as hepatocyte-specific marker in both tail vein transplantation group and hepatic lobe injection transplantation group.No significant differences of histological repair and grade of differentiation were examined between the treatments through tail vein group or direct injection into hepatic lobe group (P >0.05).