The Anti-Inflammatory Effect of Alpha-1 Antitrypsin in Hepatocyte Transplantation
The Anti-Inflammatory Effect of Alpha-1 Antitrypsin in Hepatocyte Transplantation
复制标题
Alpha-1抗胰蛋白酶在肝细胞移植中的抗炎作用
DOI：
10.1097/01.tp.0000543016.63078.e3
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发表时间：
2018
期刊：
影响因子：
6.2
通讯作者：
Lee C
中科院分区：
文献类型：
--
作者：
Lee C
体外方法：使用管环模型来分析当人肝细胞（HH）与ABO匹配的血液和4 mg/ml AAT接触时免疫应答的激活。分析血小板和白色细胞计数、补体和细胞因子表达。体内：从250 g雄性Sprague道利大鼠中分离原代大鼠肝细胞。在脾内移植2x 107个eGFP雄性肝细胞之前，对雌性同窝仔进行AAT（120 mg/kg）或盐水（对照）的尾静脉注射。分别于24 h、48 h和1 wk取肝组织行免疫组化染色和DNA提取检测Y染色体（SRY）基因表达。分离肝单核细胞（HMNC），并使用流式细胞术分析免疫细胞浸润。结果在体外：在环模型中，与仅血液对照相比，HH引起血栓形成的血小板计数显著下降（51 × 109细胞/L±15 vs 173 × 109细胞/L±8，P< 0.001）。N= 6）。含AAT和HH的环无血小板计数下降，无血栓形成（140 × 109 cell/L±13，P> 0.05）。N= 6）。AAT降低促炎细胞因子IL-1β（1.7 vs 3. 9ng/ml）、IL-6（0. 1ng/ml）和IFN-γ（0.4 vs1.与未处理的袢（913 vs 719 ng/ml）相比，具有更高的抗炎IL-1 RA浓度（N = 5，P< 0.05）。体内：在24小时和48小时，与未处理的相比，在AAT处理的肝脏中存在显著更多的eGFP肝细胞（24小时; 4.5±1.4%对1.0± 0.2%，** P< 0.01和48小时2.1± 0.5%对1.0± 0.3%，* P< 0.05）。通过使用SRY分析，AAT处理组中雄性肝细胞的植入在48小时时比对照组高10倍，在1周时高4倍（N= 3）。与未处理的对照相比，AAT在48小时降低了CD 86 + ve活化的巨噬细胞的百分比（10% vs 14. 6%）、1周（2.3% vs 6.9%）和1个月（2.1% vs 3.6%）。AAT降低促炎细胞因子的浓度; IL-1β，IL-1，IL-6和IL-2相比，对照组1h posttransplantation.ConclusionsAAT通过抑制炎性细胞因子和免疫细胞浸润提高移植肝细胞的植入。AAT可以显著延长细胞存活和功能，改善该技术用于患有肝代谢疾病的儿童的临床结果。
MethodsIn Vitro: A tubing loop model was used to analyse activation of the immune response when human hepatocytes (HH) were in contact with ABO-matched blood and 4mg/ml AAT. Platelet and white cell counts, complement and cytokine expression were analysed. In vivo: Primary rat hepatocytes were isolated from 250g male Sprague Dawley rats. Female littermates underwent tail vein injection of AAT (120mg/kg) or saline (control) prior to the intrasplenic transplantation of 2x107 eGFP male hepatocytes. At 24h, 48h and 1wk, liver was collected for immunohistochemical analysis and DNA isolated for Y chromosome (SRY) gene expression. Hepatic mononuclear cells (HMNC’s) were isolated and immune cell infiltration analysed using flow cytometry. Blood samples were used to analyse cytokine expression analysed using a Milliplex Map rat cytokine/bead panel.ResultsIn Vitro: In the loop model, HH elicited a significant drop in platelet count with thrombus formation compared to blood only controls (51x109cell/L±15 vs 173x109cell/L±8, P< 0.001. N= 6). Loops containing AAT and HH showed no drop in platelet count with no thrombus formation (140x109cell/L±13, P> 0.05. N= 6). AAT decreased pro-inflammatory cytokines IL-1β (1.7 vs3. 9ng/ml), IL-6 (0.4 vs1. 1ng/ml) and IFN-γ (0.4 vs1. 3ng/ml)(N= 5, P< 0.05) and had higher concentrations of anti-inflammatory IL-1RA compared to non-treated loops (913vs719ng/ml)(N= 5, P< 0.05). In vivo: There were significantly more eGFP hepatocytes in AAT treated livers compared to untreated at 24h and 48h (24hrs; 4.5±1.4% vs 1.0±0.2%,** P< 0.01 and 48hrs 2.1±0.5 vs 1.0±0.3%,* P< 0.05). Engraftment of male hepatocytes by analysis using SRY was 10-fold higher in the AAT treated group at 48h compared to control and 4-fold higher at 1wk (N= 3). AAT decreased the percentage of CD86+ ve activated macrophages compared to untreated controls at 48hrs (10% vs14. 6%), 1-week (2.3% vs 6.9% and 1 month (2.1% vs 3.6%). AAT decreased concentrations of pro-inflammatory cytokines; IL-1β, IL-1, IL-6 and IL-2 compared to controls 1h post transplantation.ConclusionsAAT improves engraftment of transplanted hepatocytes by inhibition of inflammatory cytokines and immune cell infiltration. AAT may significantly prolong cell survival and function, improving the clinical outcomes of the technique for children with liver based metabolic disease.