骨髓间充质干细胞对急性肝衰竭的治疗作用及机制  

Therapeutic effect and mechanism of mesenchymal stem cells on acute liver failure

作　　者：马虎成[1] 王鑫 吴旻娜 袁献温[1] 施晓雷[1] 

机构地区：[1]南京大学医学院附属鼓楼医院肝胆外科,210008 [2]南京中医药大学第一临床医学院

出　　处：《中华肝胆外科杂志》2017年第10期694-699,共6页Chinese Journal of Hepatobiliary Surgery

基　　金：国家自然科学基金（81670566,81500478）

摘　　要：目的探索骨髓间充质干细胞（MSCs）分泌的炎症相关因子及其对急性肝衰竭（ALF）的治疗作用。方法获取并鉴定sD大鼠MSCs。收集MSCs条件培养基（MSCs—CM）检测炎症相关因子。SD大鼠分为三组：ALF＋达尔伯克氏改良伊格尔培养基（DMEM）组：D－氨基半乳糖（D—Gal）诱导大鼠ALF后静脉输注DMEM 1ml；ALF＋MSCs组：诱导ALF后静脉输注MSCs 1ml（约1×10^6）；ALF＋MSCs—CM组：诱导ALF后静脉输注MSCs—CM 1ml。检测各组大鼠生化指标、4周存活率、组织病理学和炎症因子水平。评估直接输注外源性重组大鼠白介素10（IL-10）与给予抗白介素10抗体后输注MSCs-CM对ALF大鼠转氨酶的影响。检测给予IL—10后ALF大鼠肝脏磷酸化STAT3（pSTAT3）水平，评估AG490（STAT3信号通路抑制剂）能否逆转IL-10的治疗作用。结果ALF＋DMEM组、ALF＋MSCs组和ALF＋MSCs—CM组给予D—Gal第3天血清ALT分别为1709．8±372．1、865．5±52．8、964．7±414．6U／L，AST分别为4234．0±807．3、2440．8±511．9、2739．8±587．3U／L，TBil分别为79．3±10．9、43．8±7．0、61．2±6．7μg／L。三组28天存活率分别为10．0％、80．0％、70．0％；炎症因子干扰素（IFN-γ）分别为69．8±4．7、46．4±4．3、54．6±2．4pg／ml，IL-113分别为58．5±7．6、40．5±6．9、44．1±6．0pg／ml，IL-6分别为71．9±16．1、38．4±7．7、45．3±9．0pg／ml，IL-10分别为38．3±6．0、75．4±11．1、59．6±11．9pg／ml。蛋白芯片检测提示MSCs-CM表达多种炎症相关细胞因子，其中IL-10的水平最为显著。与ALF组（ALT：1709．8±372．1U／L，AST：4234．0±807．3U／L）比较，单独使用IL-10（ALT：1126．9±419．3U／L，AST：2370．8±561．2U／L）能显著降低转氨酶水平，同时给予抗IL-10抗体（ALT：1568．5±325．4U／L，AST：4043．7±819．0U／L）则能中和ISCs-CM（ALT：964．7±414．6U／L，AST：2739．8±587．3U／L）的治�Objective To explore the therapeutic effects of soluble cytokines secreted by mesenchymal stem cells （MSCs） on acute liver failure （ALF）. Methods MSCs isolated from Sprague-Dawley rats were determined by FACS analysis. Conditioned medium derived from MSCs （MSCs-CM） was collected and analyzed by a cytokine microarray. SD rats were divided into 3 groups ： （ 1 ） ALF ＋ dulbecco＇s modified eagle medium （DMEM） group： 1 ml DMEM was injected into SD rats after D-Gal administration; （2） ALF ＋ MSCs group： 1 ml MSCs （1 × 10^6） was injected into SD rats after D-Gal administration; （3） ALF ＋ MSCs-CM group： 1 ml MSCs-CM was injected into SD rats after D-Gal administration. Biochemical indicators, survival rate, histology and inflammatory factors were studied. Exogenous recombinant rat IL-10, antirat IL-10 antibody and AG490 （STAT3 signaling pathway inhibitor） were administrated to explore the therapeutic mechanism of MSCs-CM. Results The respective serum biochemical indexes of ALF ＋ DMEM group, ALF ＋ MSCs group, and ALF ＋ MSCs-CM group were： ALT （ 1 709.8 ±372.1, 865.5± 52.8, 964.7 ± 414.6U/L）, AST （4234.0 ± 807.3, 2440.8± 511.9, 2739.8 ±587.3 U/L）, andTBil （79.3 ± 10.9, 43.8 ±7.0, 61.2± 6.7 μg/L）. The survival rates of the three groups were 10.0%, 80.0%, and 70.0%, respectively. The levels of inflammatory factors in each group were IFN-γ （69.8 ± 4.7, 46.4 ± 4.3, 54.6 ±2.4pg/ml）, IL-1β （58.5 ±7.6,40.5 ± 6.9,44.1±6.0pg/ml）, IL-6 （71.9 ± 16.1, 38.4 ±7.7, 45.3 ± 9.0）, and IL-10 （38.3 ± 6.0, 75.4 ± 11.1, 59.6 ±11.9 pg/ml）. Protein microarray results suggested that MSCs-CM expresses a variety of inflammatory-related cytokines, with IL-10 levels being most pronounced. IL-10 （ALT 1 126.9 ± 419.3 U/L, AST2370.8 ±561.2 U/L） alone significantly reduced transaminase levels compared with ALF group （ALT 1 709.8± 372.1 U/L, AST 4234.0 ± 807.3 U/L）, while anti-IL-10 antibody （ALT 1568.5 ±325.4 U/L, AST4043.7 ± 819.

关 键 词：骨髓间充质干细胞 急性肝衰竭 条件培养基 免疫调节 STAT3信号通路 

分 类 号：R575.3[医药卫生—消化系统]