细粒棘球绦虫重组P29蛋白对原头蚴感染小鼠肝脏组织中TLR4及NF-кB表达的影响  

Effects of recombinant protein P29 on the TLR4 and NF-кB in the liver of mice with Echinococcus granulosus

作　　者：李菲 黎明[2] 徐士梅 朱佳佳[3] 刘成[3] 黑金娟 马锐 LI Fei;LI Ming;XU Shimei;ZHU Jiajia;LIU Cheng;HEI Jinjuan;MA Rui(School of Basic Medical Sciences,Ningxia Medical University,Yinchuan 750004,China;Department of Hepatobiliary Surgery,General Hospital of Ningxia Medical University,Yinchuan 750004,China;Science and Technology Center,Ningxia Medical University,Yinchuan 750004,China)

机构地区：[1]宁夏医科大学基础医学院,宁夏银川750004 [2]宁夏医科大学总医院肝胆外科,宁夏银川750004 [3]宁夏医科大学科技中心,宁夏银川750004

出　　处：《宁夏医学杂志》2021年第12期1066-1069,共4页Ningxia Medical Journal

基　　金：宁夏重点研发计划项目(NO2016KJHM42);宁夏大学创新、创业项目(NO.S202110752062)。

摘　　要：目的观察细粒棘球绦虫重组P29蛋白(rP29)对原头蚴感染的小鼠肝脏组织Toll样受体(TLR4)与核因子-κB(NF-κB)表达变化的影响。方法36只BALB/c的雌性小鼠,每组12只,随机分为PBS组、佐剂组和重组P29蛋白免疫组(rP29组)。PBS组和佐剂组分别注射相应的PBS和佐剂;rP29组小鼠于皮下多点注射细粒棘球绦虫重组P29蛋白(10μg/只),分别于第2、4周加强免疫;于末次免疫后2周,3组BALB/c小鼠均腹腔接种细粒棘球蚴原头蚴(1500个/只);感染后24周,摘眼球取血,处死小鼠取肝脏,切片苏木素-伊红染色(HE)观察小鼠肝脏组织的形态,Real Time PCR和Western blot法检测小鼠组织中TLR4、NF-κB-p65基因的相对表达量和蛋白表达水平。结果重组P29蛋白组肝细胞边界清楚、排列整齐,偶见炎性细胞浸润,而对照组出现较多炎性细胞浸润。重组P29蛋白组小鼠肝脏组织中的TLR4 mRNA的相对表达量为1.74±0.52,对照PBS组和佐剂组分别为1.0±0.02和1.12±0.16;重组P29蛋白组小鼠肝脏组织中的NF-κB-p65 mRNA的相对表达量为3.45±0.17,对照PBS组和佐剂组分别为1.0±0.01和2.65±0.48。重组P29蛋白免疫组TLR4和NF-κB-p65 mRNA相对表达量均高于PBS组和佐剂组。同时,重组P29蛋白组小鼠肝脏TLR4和NF-κB-p65蛋白表达量与PBS组和佐剂组相比均增高。结论重组P29蛋白能够增加细粒棘球蚴感染小鼠肝脏组织中TLR4、NF-κB-p65 mRNA和蛋白的表达。Objective To evaluate the effect of recombinant P29 protein(rP29),a 29 kDa antigen identified in hydatid cyst fluid which induces protective immunity,on TLR4 and NF-κB in the liver of mice infected with Echinococcus granulosus.Methods A group of 12 female BALB/c mice were subcutaneously immunized with 10μg of rP29 emulsified with Freund′s adjuvant,then boosted twice with two weeks interval.Another two groups of mice were given with Freund’s adjuvant or PBS only as controls.The three groups of mice were intraperitoneally challenged with 1500 protoscolex of E.granulosus per mouse two weeks after the last immunization.24 weeks after infection,the mice were euthanized and the liver tissues were collected.Results HE staining showed that the liver cells in the experimental group had clear boundaries and orderly arrangement,with occasional inflammatory cell infiltration,while more inflammatory cell infiltration was observed in the control group.The TLR4 and p65-NF-κB mRNA expression level was detected by PT-PCR in each infected mouse.The expression levels of TLR4 and p65-NF-κB mRNA in the liver tissues of mice immunized with rP29 was increased compared to PBS control mice and adjuvant control mice.The changes of the TLR4 and p65-NF-κB mRNA expression level in the liver tissues were consistent with the TLR4 and p65-NF-κBprotein expression levels detected by Western blotting.Conclusion Immunization of recombinant P29 protein could increase the expression of the TLR4 and p65-NF-κB at both mRNA and protein levels in the liver of mice infected with E.granulosus.

关 键 词：细粒棘球蚴绦虫 NF-κB-p65 TLR4 重组P29蛋白 

分 类 号：R333.4[医药卫生—人体生理学]