| 赖丽莎,陈俊伟,王劲,李丹,李征然,单鸿.骨髓间充质干细胞移植治疗大鼠急性肝衰竭[J].中国医学影像技术,2011,27(2):232~236 |
| 骨髓间充质干细胞移植治疗大鼠急性肝衰竭 |
| Effects of mesenchymal stem cells transplantation on rats with acute liver failure |
| 投稿时间：2010-10-28 修订日期：2010-11-19 |
| DOI： |
| 中文关键词: 间充质干细胞 肝衰竭,急性 氯甲基苯甲酰氨 肝再生 |
| 英文关键词:Mesenchymal stem cells Liver failure, acute CM-DiI Liver regeneration |
| 基金项目:国家自然科学基金(30770628)。 |
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| 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. |
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