NLRP3炎性复合体在异基因造血干细胞移植不同预处理方案所致肝损伤中的作用比较  

Role of NLRP3 Inflammasome in Hepatic Veno-Occlusive Disease After Allogeneic Hematopoietic Stem Cell Transplantation Undergoing Different Pre-Treatments

作　　者：黄玉金[1,2] 房婷[2] 付金玉[2] 吴进燕[2] 化静[2] 刘迷迷[2] 郭娜[2] 徐开林[2] 曾令宇[2] 

机构地区：[1]新沂市人民医院血液科,221400 [2]徐州医学院附属医院血液科实验室,221002

出　　处：《国际输血及血液学杂志》2014年第6期501-506,共6页International Journal of Blood Transfusion and Hematology

基　　金：国家自然科学基金资助项目（81070447、BK2012573、81270637）

摘　　要：目的 探讨NOD样受体蛋白（NLRP）3炎性复合体在异基因造血干细胞移植（allo-HSCT）不同预处理方案中所致肝损伤的作用.方法 选择50只无特殊病原体（SPF）级BALB/c（H-2Kd）雄性小鼠作为研究对象.将其采用计算机随机分组法分为实验组（n=40）和对照组（n=10）,再按照allo-HSCT前预处理方案不同,将实验组受鼠分为白消安/环磷酰胺（BU/CY）预处理组（n=20）和全身照射（TBI）预处理组（n=20）.实验过程中对动物的处置符合动物伦理学标准.于移植后第15天处死各组受鼠,收取其骨髓及肝脏标本,检测骨髓嵌合率和白细胞计数,并计算肝脏指数.将肝脏标本分成二部分,一部分通过苏木精-伊红（HE）染色、免疫组织化学染色及Masson 3色染色分析肝脏组织的形态学变化,进行肝静脉闭塞病（HVOD）病理学评分;另一部分采用实时荧光定量聚合酶链反应（PCR）技术检测各组受鼠肝脏组织中炎性细胞因子白细胞介素（IL）-1β、-18的相对表达量,评价细胞因子在移植后肝脏损伤中的作用;并检测细胞因子水解酶半胱氨酸天冬氨酸蛋白酶（caspase）-1与NLRP3炎性复合体在各移植受鼠肝脏组织中的相对表达量,分析其在不同预处理方案所致HVOD中的作用.结果 ①实验组受鼠骨髓细胞均成功植入,在移植后第15天,嵌合率均大于98.0％.②BU/CY预处理组受鼠并发HVOD的病理损伤程度较TBI预处理组严重,BU/CY预处理组受鼠的肝脏指数水平低于TBI预处理组;TBI预处理组受鼠中,5只受鼠的HVOD病理评分为中度,5只为重度,而BU/CY预处理组10只受鼠为重度.③BU/CY预处理组受鼠炎性细胞因子IL 1β、-18的相对表达量是TBI预处理组的13.9倍和5.5倍,两组比较,差异均有统计学意义（P＜0.05）.④BU/CY预处理组受鼠caspase-1的相对表达量为TBI预处理组的19.5倍,二者比较,差异有统计学意义（P＜0.05）;BU/CY预处理组受鼠NLRP3的相对�Objective To study the role of NOD-like domain （NLRP）3 inflammasome in hepatic venoocclusive disease （HVOD） after allogeneic hematopoietic stem cell transplantation （allo-HSCT） undergoing different pre-treatments.Methods A total of 50 specific-pathogen free （SPF） male BALB/c （H 2Kd） mice were included in this study,and randomly divided into study group （n=40） and control group （n=10） by computer.According to the differences of allo-HSCT pre-treatment programs,study group mice were divided into two groups：busulfan/cyclophosphamide （BU/CY） pre-treatment group （n =20） and total body irradiation （TBI） pre-treatment group （n=20）.The experiment was disposed with the ethical standard.On the 15th day after allo-HSCT,recipient mice in each group were sacrificed and collected bone marrow and whole liver samples,in order to detect the chimeric rate and white blood cell （WBC） count,and calculate liver index.The liver samples were divided into two parts.One part of liver samples were observed the histologic pathology by hematoxylin-eosin （HE） staining,Masson＇s trichrome staining and immunohistochemistry,and gave pathological score of HVOD to evaluate the degree of liver injury.Another part of liver samples were assessed relative expression levels of cytokine interleukin （IL）-1β,-18 by realtime fluorescence quantitative polymerase chain reaction （PCR） to study the role of inflammatory injury to HVOD.Then,liver samples were assessed relative expression levels of cysteine aspartic proteases （Caspase）-1 and NLRP3 to study role of NLRP3 in HVOD after allo-HSCT undergoing different pretreatments.Results ① Hematopoietic stem cells had been successfully transplanted into study group mice with chimeric rate over 98.0％ on the 15th day after allo-HSCT.② The pathology injury of HVOD in BU/CY pre-treatment group was more than that of TBI pre-treatment group.In BU/CY pre-treatment transplantation group,liver index was below and 10 cases of HVOD samples were diagnose

关 键 词：NLRP3蛋白 人类 细胞因子 白消安 环磷酰胺 全身照射 肝静脉闭塞病 

分 类 号：R457.7[医药卫生—治疗学]