缺氧诱导因子-1α在百草枯中毒大鼠肝脏的表达  

The expression of HIF-1 ot in liver tissues in the rat model of paraquat poisoning

作　　者：熊莺[1] 王瑞兰[1] 谢晖[1] 唐雪[1] 

机构地区：[1]上海交通大学附属第一人民医院危重病科,上海201620

出　　处：《中华急诊医学杂志》2014年第5期504-508,共5页Chinese Journal of Emergency Medicine

基　　金：国家自然科学基金（81272071）;上海市科委课题（10JC1413200）

摘　　要：目的通过观察百草枯染毒大鼠肝脏组织的病理变化及中毒后不同时间点血清转氨酶水平和肝脏组织中HIF-1α蛋白及TGF-β蛋白表达的变化，探讨HIF-1α与百草枯中毒后肝损伤的相关性。方法将48只健康SD雄性大鼠随机（随机数字法）分为对照组（6只）和染毒组（42只）：对照组大鼠予生理盐水1 mL一次性灌胃，染毒组按照50 mg/kg将20%浓度的百草枯原液用生理盐水稀释至1 mL后一次性灌胃，按灌胃后2、6、12、24、48、72 h及120 h分为7个亚组，每组6只。分别收集动脉血做血气分析及乳酸和血清转氨酶检测；肝组织切片行HE染色和Masson染色；蛋白免疫印迹法（Western blot）法检测肝组织HIF-1α和TGF-β蛋白表达。数据收集后两组之间比较采用单因素方差分析（One-Way ANOVA）。结果中毒后2 h出现转氨酶的明显升高，6 h开始出现血乳酸的明显升高，72 h开始出现低氧血症；中毒后2 h开始肝脏组织病理学表现为镜下见汇管区周围部分肝细胞变性、坏死，汇管区少量炎症细胞浸润；至中毒后12 h汇管区进一步扩大，伴大量炎性细胞浸润，并出现纤维条索样增生，且随时间延长肝细胞坏死及汇管区纤维化表现逐步向中央静脉周围延伸；中毒后2 h大鼠肝脏组织的HIF-1α与TGF-β蛋白的表达均明显升高，染毒组2、6、12、24、48、72 h及120 h各时间点与对照组的差异具有统计学意义。结论百草枯中毒后早期肝脏组织HIF-1α表达明显升高，并有可能同时上调了促纤维化因子TGF-β的表达。Objeαive To observe the levels of HIF- α and TGF-β in the liver tissue, change of serum transaminase in different phases after paraquat （PQ） toxicity and liver histopathology change in PQ- induced liver toxicity of rats models in order to analyze the relationship between HIF-1 α and hepatic toxicity induced by PQ. Methods A total of 48 healthy SD rats were randomly （ random number） assigned into 2 groups： PQ poisoning group （n = 42, 20% PQ solution, 50 mg/kg in lmL） and control group （n = 6, normal saline lmL）. Rats were separately sacrificed and liver tissue samples were harvested at 2, 6, 12, 24, 48, 72 h and 120 h after PQ administered by gastric lavage. Arterial blood gas （ABG） analysis and serum alanine transaminase were assayed. Liver tissue HE and Masson staining and immunoehemistry changes were also investigated. HIF-1 α and TGF-β levels were deteαed by Western blot technique. Results Laαic acid level was significantly higher in PQ group than that in control group at 6 h after PQ exposure （ P 〈 0. 05 ）. The level of serum alanine transaminase was increased and significantly higher if PQ group than that in control group at 2 h after PQ poisoning （P 〈0.05）. The level of HIF-lα and TGF-β protein in the liver tissue were up-regulated significantly and higher in PQ group than those in control group at 2 h after PQ exposure （P 〈 0. 05 ）. The hepatocytes necrosis and inflammatory cells infiltration were observed around portal area 2 h after PQ poisoning. At 12 h after PQ exposure, the periportal area filled with necrosis of hepatocytes and the necrosis began to extend, and huge amounts of inflammatory cell infiltration were found. Cord-like fibroplasia was found. All the histological changes were around central portal area and were enlarged gradually. Conclusions The results show that there are increased level of HIF-1α in the early stage of PQ poisoning rats. The liver fibroplasia may be associated with increasing the level of TGF-β promoted by HIF-1 α.

关 键 词：百草枯 中毒 低氧血症 急性肝损伤 缺氧诱导因子-1α(HIF-1α) 转化生长因子-β(TGF-β) 

分 类 号：R595.4[医药卫生—内科学]