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目的探讨5-脂氧合酶(5-lipoxygenase,5-LO)及其产物白三烯B4(Leukotriene B4,LTB4)在急性肝衰竭大鼠模型中的变化及其在肝脏炎性损伤中的作用。方法30只成年雄性Wistar品系大鼠分为实验对照组(6只)和肝衰竭模型组(24只)。通过腹腔注射D-氨基半乳糖及脂多糖建立大鼠急性肝衰竭模型,动态观察4、8、16、24 h各时间点大鼠肝组织的病理变化以及血清丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)、总胆红素(T-BIL)的变化。荧光定量PCR测量肝组织5-LO mRNA变化,免疫组化法及蛋白质印迹法检测5-LO蛋白表达,ELISA试剂盒检测肝组织LTB4含量。结果联合腹腔注射D-氨基半乳糖/脂多糖后,血清中ALT、AST及T-BIL逐渐升高,24 h时达高峰,ALT(3992.6±290.7)U/L、AST(10 114.7±1 014.2)U/L,T-BIL(32.9±2.0)μmmol/L,与对照组比较差异有统计学意义(P<0.01)。24 h病理改变表现为肝细胞坏死、肝索解离,小叶及汇管区大量炎性细胞浸润。5-LO mRNA表达于造模后升高,8 h表达最高;与对照组比较,肝衰竭4、8、16 h的表达差异有统计学意义(P<0.01)。造模后肝组织5-LO表达呈持续升高,24 h表达最高。LTB4含量与5-LO变化趋势相同,于造模后24 h浓度最高,为(355.4±22.2)pg/ml。结论5-LO及其产物LTB4参与急性肝衰竭的病理生理过程,并发挥重要的作用。
Objective To investigate the changes of 5-lipoxygenase (5-LO) and its product Leukotriene B4 (LTB4) in the rat model of acute hepatic failure and its role in the inflammatory injury of the liver . Methods Thirty adult male Wistar rats were divided into experimental control group (n = 6) and hepatic failure model group (n = 24). Rat models of acute hepatic failure were established by intraperitoneal injection of D-galactosamine and lipopolysaccharide. The pathological changes of rat liver tissue and serum levels of alanine aminotransferase (ALT) at 4, 8, 16 and 24 h were observed dynamically. , Aspartate aminotransferase (AST), total bilirubin (T-BIL) changes. The changes of 5-LO mRNA in liver tissue were measured by real-time PCR. The expression of 5-LO protein was detected by immunohistochemistry and Western blotting. The content of LTB4 in liver tissues was detected by ELISA kit. Results After combined intraperitoneal injection of D-galactosamine / LPS, the levels of ALT, AST and T-BIL in serum increased gradually and reached the peak at 24 h, with ALT (3992.6 ± 290.7) U / L and AST (10 114.7 ± 1 014.2 ) U / L and T-BIL (32.9 ± 2.0) μmmol / L, respectively. The difference was statistically significant compared with the control group (P <0.01). 24 h pathological changes showed hepatocellular necrosis, hepatic cord dissociation, lobular and portal area a large number of inflammatory cell infiltration. The expression of 5-LO mRNA increased at 8 h after modeling, and reached the highest at 8 h. Compared with the control group, the expression of 5-LO mRNA at 4, 8, and 16 h after hepatic failure was significantly different (P <0.01). After modeling, the expression of 5-LO in liver tissue was continuously increased, and the expression was highest at 24 h. The content of LTB4 showed the same trend as that of 5-LO, with the highest concentration of (355.4 ± 22.2) pg / ml at 24 h after modeling. Conclusion 5-LO and its product LTB4 are involved in the pathophysiological process of acute liver failure and play an important role.