中文摘要：

目的探讨骨髓间充质干细胞（MSCs）治疗大鼠急性肝衰竭（ALF）的机制,并示踪MSCs在ALF大鼠体内的分布、迁移。方法①以流式细胞仪检测hrGFP慢病毒感染的UE7T-13细胞株（hrGFP-MSCs）和氯甲基苯甲酰氨（CM-DiI）标记MSCs（MSCs-DiI）的绿色荧光蛋白阳性表达率或红色荧光染料标记阳性率。②CCK-8体外检测MSCs、hrGFP-MSCs和MSCs-DiI的细胞增殖情况。③取SD大鼠38只,建立急性肝损伤模型,30只造模成功,随机分为3组：CCL4组（A组）、CCL4/hrGFP-MSCs组（B组）和CCL4/MSCs-DiI组（C组）。B组和C组造模12 h肝内注射MSCs悬液。④分别于造模24 h、72 h、7天取肝组织以及肺组织观察移植hrGFP-MSCs、MSCs-DiI分布、迁移情况。造模后72 h,检测血清IL-10,TNF-α炎症因子水平,另取肝组织行PCNA免疫组化染色。结果①hrGFP-MSCs的hrGFP表达阳性率达99.21%,MSCs-DiI的CM-DiI标记率为99.98%。②CCK8检测示MSC-DiI、hrGFP慢病毒标记MSCs均不影响其增殖能力。③冰冻切片示CM-DiI标记MSCs可更好地示踪移植细胞,造模24 h、72 h及7天非注射部位肝组织和肺组织MSCs-DiI均可见散在的移植细胞团,而hrGFP-MSCs未见明确荧光阳性细胞。④MSCs治疗ALF大鼠模型下调了系统性炎症应答,造模72 h A组的TNF-α、IL-10水平大于B组及C组;PCNA示MSCs治疗促进了宿主肝细胞增殖。结论 CM-DiI标记MSCs能更好地示踪少量散在的移植细胞。异种移植MSCs可通过下调系统性炎症应答,促进肝细胞增殖,修复ALF大鼠肝组织。

英文摘要：

Objective To assess the therapeutic mechanisms of bone marrow mesenchymal stem cells（MSCs） on experimental rats with acute liver failure（ALF）,and to display the distribution,migration of MSCs in vivo.Methods ①hrGFP positive rate of MSCs infected with hrGFP lentivirus（hrGFP-MSCs） and CM-DiI labeling rate of MSCs labeled with CM-DiI（MSCs-DiI） were detected with flow cytometry.②MSCs,hrGFP-MSCs and MSCs-DiI were monitored for cell growth with CCK-8.③The acute liver injury models were induced in 38 SD rats.Thirty rats were established successfully and were randomly divided into CCL4 group（group A）,CCL4/hrGFP-MSCs group（group B） and CCL4/MSCs-DiI group（group C）.MSCs suspension was injected into the liver of group B and C at 12 h after modeling.④The distribution and migration of hrGFP-MSCs or MSCs-DiI in liver and lung tissue were observed at 24 h,72 h and 7 days after modeling.The serum IL-10 and TNF-α levels were detected,and liver tissue was used for PCNA immunohistochemical staining.Results ①The hrGFP positive rate of hrGFP-MSCs was 99.21% and CM-DiI labeling rate was 99.98%.②Detection in vitro with CCK-8 showed the proliferation of hrGFP-MSCs and MSC-DiI were not affected.③Frozen sections showed CM-DiI was a better cell tracing in vivo.Scattered transplanted cell clusters were seen in the non-injection site liver tissue and lung tissue in group C at 24 h,72 h,7 days after modeling,while there were not specific fluorescence-positive cells in group B.④MSCs therapy down-regulated systemic inflammation response.TNF-α and IL-10 levels of group A were higher than those of group B and C at 72 h after modeling,respectively.PCNA showed MSCs enhanced liver regeneration at 72 h after modeling.Conclusion CM-DiI is a better cell tracing for small amount of scattered cells in vivo than hrGFP with lentivirus.Xenotransplantation MSCs can down-regulate systemic inflammation response and enhance liver regeneration to repair the damage liver of rats with ALF.