泡型包虫病患者肝脏组织TGF-β1和Gadd45γ基因的表达及其在肝损伤中的作用研究  

The expression of TGF-β1 and Gadd45γ in liver tissue of patients with AE and its role in liver injury

作　　者：张传山[1] 杨舒婷[1] 毕晓娟[1] 李亮[1] 吕国栋[1] 林仁勇[1] 

机构地区：[1]新疆医科大学第一附属医院,临床医学研究院,新疆包虫病基础医学重点实验室,新疆乌鲁木齐830054

出　　处：《中国病原生物学杂志》2016年第10期908-912,共5页Journal of Pathogen Biology

基　　金：新疆维吾尔自治区自然科学基金青年基金项目(No.2014211C069)

摘　　要：目的研究泡型包虫病患者肝脏组织TGF-β1和Gadd45γ基因的表达及其在肝损伤中的作用,探讨其临床意义。方法采集23例AE患者肝脏手术标本,Trizol法提取RNA,反转录cDNA后采用实时荧光定量PCR检测TGF-β1、Gadd45γ、p53和p21基因的表达。体外培养肝细胞HL-7702,用重组TGF-β1(1ng/ml)细胞因子或匀浆虫体蛋白(1mg/ml)刺激30min、1h、24h和48h,分别收集细胞,采用实时荧光定量PCR检测TGF-β1、Gadd45γ、p53和p21基因的表达。结果 AE患者病灶旁肝脏组织TGF-β1、p53和p21mRNA的相对表达量分别为1.93±1.34、1.29±0.62和1.50±1.10,与远端肝组织1.00±0.00比较差异有统计学意义(P<0.05);Gadd45γmRNA为1.24±0.96,与远端组织比较差异无统计学意义(P>0.05)。Pearson法分析TGF-β1表达分别与Gadd45γ、p53和p21呈正相关(r值分别为0.5257、0.5287、0.4605,P<0.05)。用重组细胞因子TGF-β1(1ng/ml)或匀浆虫体蛋白(1mg/ml)刺激30min、1h、24h和48h,均能促进肝细胞HL-7702中TGF-β1、Gadd45γ、p53和p21基因的表达上调。结论在肝泡球蚴病晚期,炎症性细胞因子TGF-β1可能通过激活非经典型MAPK(Mitogen Activated Protein Kinase,JNK和p38)信号转导途径和细胞周期关键调控因子(Gadd45γ,p53,p21)调控肝脏细胞的生长抑制或凋亡,造成宿主肝脏的病理性损伤。Objectives To study the expression of TGF-β1and Gadd45γin liver tissue of patients with AE and its role in liver injury and to explore its clinical significance. Methods Total RNA was extracted from liver specimens from 23 patients with AE using the TRIzol method,and RNA was then reverse-transcribed into cDNA.HL-7702 cells were maintained in Dulbecco＇s Modified Eagle Medium containing Nutrient Mixture F-12（DMEM/F-12）supplemented with 10%fetal calf serum.Cells were stimulated with recombinant TGF-β1（1ng/ml）cell factor or E.multilocularis proteins（1mg/ml）for 30 min,1h,24 h,and 48 h.The levels of TGF-β1,Gadd45γ,p53,and p21 expression were detected with quantitative real-time PCR（qRT-PCR）. Results The relative level of TGF-β1mRNA expression in liver tissue adjacent to the lesion was 1.93±1.34,the relative level of p53 mRNA expression was 1.29±0.62,and the relative level of p21 mRNA expression was 1.50±1.10.These levels were significantly higher（P〈0.05）than those in distal liver tissue（1.00±0.00）.However,the relative level of Gadd45γmRNA expression was 1.24±0.96,and this level did not differ significantly from that in distal liver tissue（P〉0.05）.Pearson analysis indicated that TGF-β1expression was positively correlated with Gadd45γ,p53,and p21expression（r=0.5257,P〈0.05;r=0.5287,P〈0.05;r=0.4605,P〈0.05）.In addition,stimulation with the recombinant cytokine TGF-β1（1ng/ml）or E.multilocularis proteins（1mg/ml）for 30 min,1h,24 h,and 48 hpromoted the up-regulated expression of TGF-β1,Gadd45γ,p53,and p21 mRNA in HL-7702 liver cells. Conclusion In the later stage of E.multilocularisinfection,TGF-β1may inhibit the growth or regulate the apoptosis of liver cells by activating an atypical MAPK signalling pathway（mitogen-activated protein kinase,JNK,and p38）and cell cycle regulatory factors（Gadd45γ,p53,and p21）,ultimately causing pathological damage to the liver.

关 键 词：泡型包虫病 转化生长因子β1 生长抑制和DNA损伤基因45γ 肝损伤 生长抑制或凋亡 

分 类 号：R532.32[医药卫生—内科学]