摘要
目的:骨髓间充质干细胞分泌的因子(mesenchymal stem cell-derived molecules,MSC-CM)对急性肝衰竭小鼠高迁移率族蛋白B1(high mobility group box1,HMGB1)肝细胞胞浆移位和释放的影响.方法:D-氨基半乳糖(D-galactosamine,D-GaIN)和脂多糖(lipopolysaccharides,LPS)诱导建立Balb/c小鼠急性肝衰竭模型;贴壁筛选法培养纯化小鼠BMSCs,传至第2-3代更换0.05%胎牛血清培养液24h获得MSC-CM.36只健康Balb/c小鼠随机均分为肝衰竭对照组和MSC-CM治疗组.Kaplan-meier法进行生存分析,生化检测1wk内不同时间点各实验组丙氨酸转氨酶(alanine aminotransferase,ALT)/谷草转氨酶(aspartate transaminase,AST),24h取肝脏进行肝脏病理检测.ELISA检测血清HMGB1、白介素-1(interleukin-1,IL-1、肿瘤坏死因子-(tumor necrosis factor-,TNF-、IL-6和IL-10水平,免疫组织化学分析HMGB1的肝细胞表达和胞浆移位.结果:MSC-CM治疗组1wk生存率为88.9%,显著高于对照组的16.7%(P<0.05),MSC-CM治疗组24h的ALT/AST峰值显著低于对照组(P<0.01).MSC-CM治疗组在6、12和24hHMGB1水平以及24h的TNF-、IL-1、IL-6水平显著低于肝衰竭对照组(P<0.01),而抗炎因子IL-10显著高于对照组(P<0.01).MSC-CM治疗组肝脏炎症坏死和肝细胞HMGB1胞浆移位较对照组明显减轻.结论:MSC-CM治疗抑制急性肝衰竭肝细胞HMGB1胞浆移位和释放,减轻肝脏炎症反应,降低死亡率.
AIM： To explore the influence of mesenchymal stem cell-derived molecules （MSC-CM） on cyto- plasmic translocation of HMGB1 in hepatocytes in an animal model of acute live failure.METHODS： D-galactosamine （D-GaIN） and lipo- polysaccharide （LPS） were used to induce acute liver failure in Balb/c mice. BMSCs were cultured,purified and passaged to 2-3 generations, and the medium was replaced with DMEM containing 0.05% fetal bovine serum. After further culture for 24 h, the medium containing MSC-CM was obtained. Thirty-six healthy Balb/c mice were randomly divided into two groups： an acute liver failure group （control group） and a MSC-CM treat- ment group. The Kaplan-Meier method was used for survival analysis. Serum ALT/AST levels were measured at different time points, and liver tissue pathological examination was performed. Serum levels of HMGB1, interleukin-α（IL-I~）, tumor necrosis factor-α （TNF-α）, IL-6 and IL-10 were de- tected by ELISA. HMGB1 expression and cytoplas- mic translocation in hepatocytes were detected by immunohistochemistry.RESULTS： The survival rate was significantly higher in the MSC-CM treatment group than in the control group （89.9% vs 50%, P 〈 0.05）. MSC-CM treatment decreased the peak levels of AST/ALT at 24 h compared to the control group （P 〈 0.01）. Serum levels of HMGB1 at 6, 12, and 24 h and TNF-a, IL-113, IL-6 at 24 h were also decreased in the MSC-CM treatment group （all P 〈 0.01）; however, IL-10 level was increased in MSC-CM treatment group （P 〈 0.01）. Inflam- mation or necrosis in liver tissue and cytoplasm translocation of HMGB1 in hepatocytes were decreased in the MSC-CM treatment group com- pared to the control group （both P 〈 0.01）.
出处
《世界华人消化杂志》
CAS
北大核心
2013年第21期2009-2015,共7页
World Chinese Journal of Digestology
基金
国家自然科学基金资助项目
No.81160065
江西省科技支撑基金资助项目
No.2009JX0096
南昌市科技局重点基金资助项目
No.洪财政[2012]37号社会发展支撑计划-10~~
关键词
骨髓间充质干细胞分泌因子
高迁移率族蛋白B-1
胞浆移位
急性肝衰竭
MSC-CM treatment decreasescytoplasm translocation of HMGB1 in hepa-tocytes and attenuates liver inflammation andmortality in acute liver failure.
