中华血液学杂志
中華血液學雜誌
중화혈액학잡지
Chinese Journal of Hematology
2014年
8期
684-688
,共5页
黄玉金%付金玉%吴进燕%房婷%化静%刘迷迷%郭娜%徐开林%曾令宇
黃玉金%付金玉%吳進燕%房婷%化靜%劉迷迷%郭娜%徐開林%曾令宇
황옥금%부금옥%오진연%방정%화정%류미미%곽나%서개림%증령우
造血干细胞移植%肝脏损伤%细胞因子%基因,NLRP3
造血榦細胞移植%肝髒損傷%細胞因子%基因,NLRP3
조혈간세포이식%간장손상%세포인자%기인,NLRP3
Hematopoietic stem cell transplantation%Liver damage%Cytokine%Gene,NLRP3
目的 探索NOD样受体蛋白3(NLRP3)炎性复合体表达与小鼠异基因造血干细胞移植(allo-HSCT)肝损伤的关系.方法 建立allo-HSCT小鼠模型,流式细胞术检测受鼠骨髓嵌合率;通过HE染色、免疫组化染色及Masson染色观察肝组织病理形态,通过病理损伤评分评价肝损伤程度;用Western blot法检测肝脏组织炎性细胞及NLRP3炎性复合体表达水平;用实时荧光定量PCR法检测肝组织NLRP3炎性复合体组分相关基因及细胞因子IL-1β、IL-18 mRNA表达水平.结果 allo-HSCT后第10天(+10 d),供鼠造血干细胞基本上完全植入,嵌合率大于97％,同时肝脏病理形态显示有肝损伤,+15d肝损伤最重,随后逐渐减轻;在肝损伤过程中伴有IL-1β、IL-18 mRNA表达水平增高,+15d时二者表达量分别为正常组的(1.19±0.40)倍和(1.64±0.76)倍;在肝损伤最严重时伴有caspase-1mRNA表达水平增高,为正常组的(3.51±0.46)倍;移植后肝组织细胞NLRP1、NLRP3、NLRC4、NLRP5mRNA均有一定程度的表达,其中NLRP3表达水平最高,其mRNA及蛋白表达水平与肝损伤程度同步,+15d时mRNA表达量为正常组的(2.91±0.41)倍.结论 NLRP3在allo-HSCT小鼠表达水平明显增高,可能为移植后肝脏炎症性损伤的因素之一.
目的 探索NOD樣受體蛋白3(NLRP3)炎性複閤體錶達與小鼠異基因造血榦細胞移植(allo-HSCT)肝損傷的關繫.方法 建立allo-HSCT小鼠模型,流式細胞術檢測受鼠骨髓嵌閤率;通過HE染色、免疫組化染色及Masson染色觀察肝組織病理形態,通過病理損傷評分評價肝損傷程度;用Western blot法檢測肝髒組織炎性細胞及NLRP3炎性複閤體錶達水平;用實時熒光定量PCR法檢測肝組織NLRP3炎性複閤體組分相關基因及細胞因子IL-1β、IL-18 mRNA錶達水平.結果 allo-HSCT後第10天(+10 d),供鼠造血榦細胞基本上完全植入,嵌閤率大于97％,同時肝髒病理形態顯示有肝損傷,+15d肝損傷最重,隨後逐漸減輕;在肝損傷過程中伴有IL-1β、IL-18 mRNA錶達水平增高,+15d時二者錶達量分彆為正常組的(1.19±0.40)倍和(1.64±0.76)倍;在肝損傷最嚴重時伴有caspase-1mRNA錶達水平增高,為正常組的(3.51±0.46)倍;移植後肝組織細胞NLRP1、NLRP3、NLRC4、NLRP5mRNA均有一定程度的錶達,其中NLRP3錶達水平最高,其mRNA及蛋白錶達水平與肝損傷程度同步,+15d時mRNA錶達量為正常組的(2.91±0.41)倍.結論 NLRP3在allo-HSCT小鼠錶達水平明顯增高,可能為移植後肝髒炎癥性損傷的因素之一.
목적 탐색NOD양수체단백3(NLRP3)염성복합체표체여소서이기인조혈간세포이식(allo-HSCT)간손상적관계.방법 건립allo-HSCT소서모형,류식세포술검측수서골수감합솔;통과HE염색、면역조화염색급Masson염색관찰간조직병리형태,통과병리손상평분평개간손상정도;용Western blot법검측간장조직염성세포급NLRP3염성복합체표체수평;용실시형광정량PCR법검측간조직NLRP3염성복합체조분상관기인급세포인자IL-1β、IL-18 mRNA표체수평.결과 allo-HSCT후제10천(+10 d),공서조혈간세포기본상완전식입,감합솔대우97％,동시간장병리형태현시유간손상,+15d간손상최중,수후축점감경;재간손상과정중반유IL-1β、IL-18 mRNA표체수평증고,+15d시이자표체량분별위정상조적(1.19±0.40)배화(1.64±0.76)배;재간손상최엄중시반유caspase-1mRNA표체수평증고,위정상조적(3.51±0.46)배;이식후간조직세포NLRP1、NLRP3、NLRC4、NLRP5mRNA균유일정정도적표체,기중NLRP3표체수평최고,기mRNA급단백표체수평여간손상정도동보,+15d시mRNA표체량위정상조적(2.91±0.41)배.결론 NLRP3재allo-HSCT소서표체수평명현증고,가능위이식후간장염증성손상적인소지일.
Objective To explore the function ofnucleotide-binding domain (NOD)-like receptor protein 3 (NLRP3) inflammsomes in liver damage after allogeneic hematopoietic stem cell transplantation (allo-HSCT).Methods The study presented a murine (BALB/c-based) model of allo-HSCT.Chimera rate was measured by flow cytometry.The hematoxylin-eosin,Masson' s trichrome,immunohistochemistry staining were used to observe the pathology changes in liver,then measured the degree of liver damage.Inflammation cells and NLRP3 were measured by Western blot,cytokines IL-1β,IL-18 and NLRP3 related genes were tested with real-time quantitative polymerase chain reaction (q-PCR).Results Hematopoietic stem cells had been successfully transplanted,the chimera rate was geater than 97％ on the 10th day.Liver damage occurred after allo-HSCT and suffered infiltration of inflammation cells,which reached the peak on day 15,then moved to moderate; the cytokines IL-1β,IL-18 had the similar trend with liver injury,and reached the highest level on day 15,their mRNA expressions increased by (1.19±0.40) fold and (1.64±0.76) fold,respectively; Meanwhile,caspase-1 had the similar trend,its mRNA expression increased by (3.51±0.46) fold on day 15; the inflammasomes NLRP1,NLRP3,NLRC4 and NLRP5 expressed in liver on day 15 of post-allo-HSCT,and NLRP3 inflammasome expressed highest among them.The mRNA and protein level of NLRP3 inflammasomes were kept with the serious degree of the liver damage,its mRNA expression increased by (2.91±0.41) fold on day 15.Conclusion NLRP3 inflammsome expressed in liver injury during allo-HSCT in mice,and may be one of the important factors contributed to liver injury.
