老鼠簕生物碱A对小鼠药物性肝损伤的保护作用

目的:观察老鼠簕生物碱A(AAIA)对对乙酰氨基酚引起肝损伤模型的影响。方法:将小鼠随机分成正常组、模型组、联苯双酯组(联苯双酯,150 mg·kg~(-1))及AAIA高、中、低剂量(200,100,50 mg·kg~(-1))组,每组10只,连续灌胃给药10 d,末次给药后禁食不禁水8 h,除正常组外,其余各组给予腹腔注射275 mg·kg~(-1)对乙酰氨基酚,诱导小鼠急性肝损伤模型。6 h后,进行眼球取血,称量小鼠体质量、肝脏、脾脏、肾脏、胸腺质量,计算相应脏器指数,使用试剂盒检测血清中丙氨酸氨基转移酶(ALT),天冬氨酸氨基转移酶(AST)的含量,紫外分光光度法测定肝匀浆中一氧化氮(NO),诱导型一氧化氮合酶(i NOS)的含量,蛋白免疫印迹法(Western blot)测定细胞外调节蛋白激酶(ERK1/2),磷酸化细胞外调节蛋白激酶(p-ERK1/2)的表达。结果:与正常组比较,模型组的肝脏指数,血清中AST,ALT,肝组织匀浆NO,iNOS水平及p-ERK1/2在肝脏中的蛋白表达显著增高(P 0. 01);与模型组比较,各给药组均能降低对乙酰氨基酚急性肝损伤小鼠肝脏指数,血清AST,ALT水平,肝组织匀浆中NO,iNOS的产生,肝脏p-ERK1/2蛋白表达(P 0. 01);所有注射对乙酰氨基酚组的脾脏、肾脏及胸腺无明显变化,差异无统计学意义。结论:AAIA可能通过降低AST,ALT的水平,下调NO,iNOS的表达,减少p-ERK1/2的表达,对小鼠药物性肝损伤起到保护作用。

Protective Effect of Alkaloid A from Acanthi Ilicifolii Herba seu Radix on Drug-induced Liver Injury in Mice

Objective:To observe the effect of alkaloid A from Acanthi Ilicifolii Herba seu Radix(AAIA) on liver injury model caused by acetaminophen. Method:Mice were randomly divided into normal group, model group, positive drug group (bifendate, 150 mg·kg^-1) and high, medium and low-dose AAIA groups (200, 100, 50 mg·kg^-1), with 10 in each group. They were given drugs by gavage for 10 days, and fasted for 8 hours after the last administration. Except the normal group, the other groups were intraperitoneally injected with 275 mg·kg^-1 acetaminophen to induce acute liver injury model in mice. Six hours later, blood was taken from the eyeball. The body, liver, spleen, kidney and thymus were weighed, and then the corresponding organ indexes were calculated. The kits were used to detect the contents of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) in serum. The contents of nitric oxide (NO) and inducible nitric oxide synthase (iNOS) in liver homogenate were determined by ultraviolet spectrophotometry, and the expressions of extracellular regulated protein kinases (ERK1/2) and phosphorylated extracellular regulatory protein kinase (p-ERK1/2) were determined by Western blot. Result:Compared with the normal group, the liver index, serum AST and ALT levels, the production of NO and iNOS in liver homogenate, the expression of p-ERK1/2 protein in liver of the model group increased significantly (P<0.01). Compared with the model group, all of drug groups could reduce the liver index of mice with acute liver injury induced by acetaminophen, the levels of serum AST and ALT, the production of NO and iNOS in liver and homogenate and the expression of p-ERK1/2 protein in liver (P<0.01). however, it had no significant effect on spleen, kidney and thymus of all acetaminophen injection groups. Conclusion:AAIA may protect mice from drug-induced liver injury by reducing AST and ALT levels, down-regulating the expressions of NO and iNOS, and reducing the expression of protein p-ERK1/2.