小鼠肝星状细胞(HSC)中ATF5促进TRAIL基因的表达及其对同种异体胰岛移植物的保护作用  

ATF5 promotes the expression of TRAILgene in murine hepatic stellate cells(HSCs) and its protective effects on allogeneic islets allografts

作　　者：黄文海 陈宗祐 项建斌 李震洋 顾晓冬 HUANG Wen-haI;CHEN Zong-you;XIANG Jian-bin;LI Zhen-yang;GU Xiao-dong(Department of General Surgery,Huashan HospitalFudan University Shanghai 200040..China)

机构地区：[1]复旦大学附属华山医院普外科,上海200040

出　　处：《复旦学报（医学版）》2018年第6期781-787,共7页Fudan University Journal of Medical Sciences

摘　　要：目的探讨小鼠肝星状细胞(hepatic stellate cells,HSCs)中转录激活因子5 (activating transcription factor 5,ATF5)促进肿瘤坏死因子相关凋亡诱导配体(tumor necrosis factor-related apoptosis-inducing ligand,TRAIL)的表达及其对同种异体胰岛移植物的保护作用。方法半定量RT-PCR检测HSCs中ATF5 mRNA、TRAIL mRNA的表达水平,流式细胞术检测HSCs表面TRAIL蛋白质水平。慢病毒(LV-ATF5-RNAi)感染HSCs,针对ATF-5基因进行RNA干扰,半定量RT-PCR检测慢病毒感染的HSCs中ATF5 mRNA及TRAIL mRNA水平。单向混合淋巴细胞反应(mixed lymphocyte reaction,MLR)观察HSCs和慢病毒感染的HSCs对T细胞的增殖反应。分别将单纯胰岛、胰岛联合HSCs、胰岛联合慢病毒感染的HSCs移植到小鼠的肾包膜下,比较移植胰岛的存活时间。结果γ-干扰素(interferon-γ,IFN-γ)上调HSCs中ATF5 mRNA和TRAIL mRNA水平(P<0.01),IFN-γ亦可上调HSCs表面TRAIL蛋白质水平。慢病毒(LV-ATF5-RNAi)感染HSCs后,HSCs中ATF5 mRNA水平明显下降(P<0.01),TRAIL mRNA水平亦下降(P<0.05)。IFN-γ作用于感染慢病毒的HSCs后,HSCs中ATF5 mRNA和TRAIL mRNA水平亦增加(P<0.01),但明显低于未被慢病毒感染的HSCs(P<0.01)。HSCs可明显抑制T细胞增殖反应(P<0.01),慢病毒感染的HSCs亦能抑制T细胞增殖反应(P<0.05),但抑制作用明显低于未被慢病毒感染的HSCs(P<0.01)。单纯胰岛移植组(A组)移植胰岛中位存活时间为11天;胰岛联合HSCs移植组(B组)移植胰岛中位存活时间为88天,B组胰岛移植物存活时间明显长于A组(P<0.01);胰岛联合慢病毒感染的HSCs移植组(C组)移植胰岛中位存活时间为18天,C组胰岛移植物存活时间长于A组(P<0.01),但明显短于B组(P<0.01)。结论小鼠HSCs中ATF5可促进TRAIL基因表达,表达TRAIL的HSCs可以抑制T细胞的增殖反应,此HSCs与胰岛细胞联合移植,明显延长小鼠同种异体胰岛移植物存活时间。Objective To investigate the role of activating transcription factor 5(ATF5)on promoting the expression of tumor necrosis factor-related apoptosis-inducing ligand(TRAIL)in murine hepatic stellate cells(HSCs)and its protective effects in allogeneic islets allografts. Methods The levels of ATF5 mRNA and TRAIL mRNA in HSCs were detected by semi-quantitative RT-PCR,the level of TRAIL protein on HSCs was detected by flow cytometry.RNA interference of ATF-5 gene was carried out in HSCs through infecting lentivirus(LV-ATF5-RNAi),the levels of ATF5 mRNA and TRAIL mRNA in HSCs infected with LV-ATF5-RNAi were detected by semi-quantitative RTPCR.Effects of HSCs and HSCs infected with LV-ATF5-RNAi on T cells proliferation were observed in one-way mixed lymphocyte reaction(MLR).We compared the survival time among islets alone,islets mixed with HSCs,and islets mixed with HSCs infected with LV-ATF5-RNAi transplanted to subcapsular space of recipient kidney. Results IFN-γupregulated the levels of ATF5 mRNA and TRAIL mRNA in HSCs(P<0.01),IFN-γalso upregulated the level of TRAIL protein on HSCs.The level of ATF5 mRNA was significantly decreased in HSCs infected with LV-ATF5-RNAi(P<0.01),the level of TRAIL mRNA was also decreased in HSCs infected with LV-ATF5-RNAi(P<0.05).The levels of ATF5 mRNA and TRAIL mRNA were also increased when HSCs infected with LV-ATF5-RNAi were exposed to IFN-γ(P<0.01),but much lower than that in HSCs(P<0.01).HSCs could significantly inhibit the proliferation of T cells(P<0.01).HSCs infected with LV-ATF5-RNAi could also inhibit the proliferation of T cells(P<0.05),but the inhibitory effect was significantly lower than that of HSCs(P<0.01).Median survival time in islets transplantation group(group A)was 11 days.Median survival time in islets mixed with HSCs transplantation group(group B)was 88 days,the survival time of islets grafts in group B was significantly longer than that in group A(P<0.01).Median survival time in islets mixed with HSCs infected with LV-ATF5-RNAi transplantation group(group C)was

关 键 词：肝星状细胞(HSC) 转录激活因子5(ATF5) 肿瘤坏死因子相关凋亡诱导配体(TRAIL) 胰岛移植 小鼠 

分 类 号：R458[医药卫生—治疗学]