Zuo He-ping, Zhao Yuan-yuan, Qiu Qiao-meng, Lu Zhong-qiu, Hong Guang-liang, Li Meng-fang
Department of Emergence, First Affiliated Hospital, Wenzhou Medical College, Wenzhou 325000, China.
Zhonghua Yu Fang Yi Xue Za Zhi. 2011 Aug;45(8):702-6.
To investigate the influence of genetic polymorphism in NF-E2-related factor-2 (nrf2) gene promoter locus at 336 in alcoholic liver disease (ALD) with Vibrio vulnificus (VV) sepsis.
Through the simple random sampling method, C57B6 male mice were divided into normal feeding group (group A, 10 mice), alcoholic liver disease group (group B, 10 mice), normal feeding group infected with VV through intraperitoneal injection (group C, 8 mice), alcoholic liver disease group infected with VV (group D, 110 mice). Through gene sequencing method, nrf2 gene promoter 336 polymorphism in D group was analyzed and grouped into: non-mutation group (336T) (group D1, 7 mice) and mutation group (336C) (group D2, 10 mice). Through RT-PCR, Western-blotting and ELISA method, expressions of nrf2, tumor necrosis factor-α (TNF-α), interleukin-10 (IL-10), high mobility group protein 1 (HMGB(1)) gene and protein of liver were measured. The pathological changes in liver were recorded with light microscope.
After infected with VV for 48 hours for A, B, C, D1, D2 group, the expression medians of nrf2 mRNA in liver were 0.115, 0.173, 0.211, 0.764, 0.352, respectively (χ(2) = 40.64, P < 0.05), the expression medians of IL-10 mRNA in liver were 0.338, 0.637, 1.002, 1.825, 1.403, respectively (χ(2) = 41.05, P < 0.05), the expression medians of TNF-α mRNA in liver were 0.140, 0.254, 0.372, 0.399, 0.699, respectively (χ(2) = 38.16, P < 0.05), the expression medians of HMGB(1) mRNA in liver were 0.230, 0.410, 0.668, 0.508, 1.021, respectively (χ(2) = 31.45, P < 0.05). After infected with VV 48 hours for mice in A, B, C, D1, D2 group, the expression medians of nrf2 protein in liver were 0.908, 1.461, 2.061, 3.982, 2.243, respectively (χ(2) = 33.72, P < 0.05), the expression medians of IL-10 protein in liver were 13.97, 22.54, 30.14, 57.98, 41.53, respectively (χ(2) = 37.31, P < 0.05), the expression medians of TNF-α protein in liver were 114.07, 142.94, 175.44, 174.60, 266.11, respectively (χ(2) = 32.29, P < 0.05), the expression medians of HMGB(1) protein in liver were 2.01, 6.05, 9.62, 6.24, 12.89, respectively (χ(2) = 36.94, P < 0.05). Compared with group A, there were large amount of fat drops, fatty changes in group B, inflammatory cell infiltration, disorder of hepatic cell in group C, and extension of hepatic duct and vein, edema of liver cells and disorder of hepatic cells in group D.
The nrf2 gene promoter of T336C mutation in C57B6 mouse of ALD can significantly decrease the expression of nrf2, and intensify organ inflammation and damage when they were infected by VV.
探讨酒精性肝病（ALD）患者核因子E2相关因子2（nrf2）基因启动子区域336位点基因多态性对创伤弧菌（VV）败血症的影响。
采用简单随机抽样法，将C57B6雄性小鼠分为正常饲养组（A组，10只）、酒精性肝病组（B组，10只）、腹腔注射VV感染的正常饲养组（C组，8只）、酒精性肝病感染VV组（D组，110只）。采用基因测序法分析D组nrf2基因启动子336位点多态性，并分为：非突变组（336T）（D1组，7只）和突变组（336C）（D2组，10只）。采用RT-PCR、Western-blotting和ELISA法检测肝脏中nrf2、肿瘤坏死因子-α（TNF-α）、白细胞介素-10（IL-10）、高迁移率族蛋白1（HMGB(1)）基因及蛋白表达。用光镜记录肝脏病理变化。
A、B、C、D1、D2组小鼠腹腔注射VV 48小时后，肝脏中nrf2 mRNA表达中位数分别为0.115、0.173、0.211、0.764、0.352（χ(2)=40.64，P<0.05）；IL-10 mRNA表达中位数分别为0.338、0.637、1.002、1.825、1.403（χ(2)=41.05，P<0.05）；TNF-α mRNA表达中位数分别为0.140、0.254、0.372、0.399、0.699（χ(2)=38.16，P<0.05）；HMGB(1) mRNA表达中位数分别为0.230、0.410、0.668、0.508、1.021（χ(2)=31.45，P<0.05）。A、B、C、D1、D2组小鼠腹腔注射VV 48小时后，肝脏中nrf2蛋白表达中位数分别为0.908、1.461、2.061、3.982、2.243（χ(2)=33.72，P<0.05）；IL-10蛋白表达中位数分别为13.97、22.54、30.14、57.98、41.53（χ(2)=37.31，P<0.05）；TNF-α蛋白表达中位数分别为114.07、142.94、175.44、174.60、266.11（χ(2)=32.
