摘要目的 观察高迁移率蛋白B1-A盒(HMGB1-Abox)蛋白对急性肝衰竭小鼠肝组织表达高迁移率蛋白(HMG)B1的影响及其对炎性因子分泌的抑制作用.方法 利用克隆载体构建小鼠PET28a-HMGB1-Abox原核质粒,转化大肠杆菌BL21,以异丙基-β-D-硫代半乳糖苷诱导后经镍离子亲和层析柱亲和纯化得到目的蛋白;用脂多糖和D-氨基半乳糖胺制备小鼠急性肝衰竭模型,对照组腹腔注射等渗盐水,实验组腹腔注射重组HMGB1-Abox蛋白,于2、4、6、8、12、24 h免疫组织化学检测其肝组织中HMGB1表达的变化情况,逆转录-PCR检测其HMGB1-mRNA变化趋势以及血清中肿瘤坏死因子α、白细胞介素-1β的含量.两组间数据采用配对t检验法比较.结果 构建了PET28a-HMGB1-Abox原核质粒并纯化出1.4×10~4的目的蛋白,实验组肝组织中肝脏坏死情况明显好转,HMGB1和HMGB1-mRNA的表达较对照组降低且表达时间延后;实验组肿瘤坏死因子α于4 h开始上升至(102.46±2.51)pg/ml,12 h达到峰值(334.12±39.13)pg/ml,对照组于8h达到峰值(473.42±22.99)pg/ml,两组比较,t=-11.387,P<0.05,差异有统计学意义.实验组白细胞介素-1β自2h起随时延长逐渐增加,在6h达到峰值(341.31±25.01)pg/ml,对照组于2 h达到峰值(724.49±34.24)pg/ml,两组比较,t=-7.999,P<0.05,差异有统计学意义.结论 重组HMGB1-Abox蛋白可以显著减少急性肝衰竭时肝组织HMGB1的表达和分泌,有效的阻断HMGB1对其他早期炎性因子的促进作用.

abstractsObjective To evalute the effect of recombinant HMGB1 A box protein in mouse with acute hepatic failure.Method Acute hepatic failure was induced by D-galactosamine and lipopolysaccharide in mice.After injection with saline(control)or recombinant HMGB1 A box proteins,the expression of HMGB1 in liver tissues was detected by immunohistochemisty and RT-PCR,and the serum TNF α and IL1β were quantified.Results rHMGB1-Abox protein alleviated the acute liver damage,rHMGB 1-Abox protein treatment decreased the expression of HMGB1 in liver tissues and reduced the serum levels of TNF α and IL-1β.Conclusion rHMGB1-Abox protein can alleviate the acute liver damage and inhibit the expression of HMGB1.