蓝莓对四氯化碳所致急性肝损伤大鼠肝组织GCLCmRNA及蛋白表达的影响

摘要

目的：探讨蓝莓对四氯化碳（CCl4）所致急性肝损伤大鼠肝组织γ‐谷氨酰半胱氨酸合成酶（γ‐GCS）催化亚基（GCLC）mRNA及蛋白表达的影响。方法将50只雄性Wistar大鼠分为5组，即空白组、模型组、蓝莓汁低剂量预防组（蓝低组，10．0g／kg）、蓝莓汁高剂量预防组（蓝高组，20．0g／kg）、联苯双酯预防组（联苯双酯组，0．15g／kg），每组10只，连续灌胃7d，1次／天，其后用CCl4复制大鼠急性肝损伤模型。光镜下观察大鼠肝脏病理学变化，全自动生化分析仪测定血清丙氨酸氨基转移酶（ALT）及天冬氨酸氨基转移酶（AST）活性，酶联免疫吸附法检测肝匀浆中丙二醛（MDA）、过氧化氢酶（CAT）、超氧化物歧化酶（SOD）、还原型谷胱甘肽（GSH）活力及含量，分别采用反转录PCR（RT‐PCR）、免疫组织化学法、Westernblot检测大鼠肝组织中GCLCmRNA及蛋白表达。结果蓝低组、蓝高组和联苯双酯组大鼠肝细胞变性及坏死程度显著轻于模型组（P＜0．05），血清ALT、AST及肝匀浆MDA均显著低于模型组（P＜0．01），而肝匀浆CAT、SOD、GSH均显著高于模型组（P＜0．01），肝组织GCLCmR‐NA及蛋白表达均显著高于模型组（P＜0．01）。3组相比，蓝高组与联苯双酯组效果相似，略优于蓝低组。结论蓝莓对CCl4诱导的大鼠急性肝损伤有一定预防作用，可能与上调大鼠肝脏GCLC表达，减轻氧化应激性肝损伤有关。%Objective To observe the effect of blueberry on the mRNA and protein expression of GCLC in rats with acute hepat‐ic injury induced by carbon tetrachloride (CCl4 ) .Methods A total of 50 male Wistar rats were randomly divided into the normal control group (A) ,the model group (B) ,the low dose blueberry group (10 .0 g/kg ,C) ,the high dose blueberry group (20 .0 g/kg , D) ,and the bifendate group (0 .15 g/kg ,E) ,and there were 10 rats in each group .Each group was respectively administered with corresponding drugs by gastrogavage ,once per day for 7 days .Then the rats in latter four groups were given CCl4 to induce acute hepatic injury model .The morphological changes of the liver tissue were observed by HE stain .Detection of ALT and AST in serum were performed by an automatic biochemical analyser .The concentrations of MDA ,CAT ,SOD and GSH in the liver homogenate were determined by enzyme linked immunosorbent assay .The mRNA and protein expression of GCLC in the liver tissue were deter‐mined by RT‐PCR ,immunohistochemical assay ,and Western blot respectively .Results Compared with group B ,the degree of he‐patic degenerative necrosis were significantly improved(P<0 .05) ,the activity of ALT and AST in serum and content of MDA in the liver homogenate were significantly lower in group C ,Group D and Group E(P<0 .01) ,the content of CAT ,SOD and GSH in the liver homogenate were significantly higher in Group C ,Group D and Group E (P<0 .01) ,and the mRNA and protein expression of GCLC were significantly higher in Group C ,Group D and Group E (P<0 .01) .Among group C ,group D and group E ,group E had similar effect with group D ,and both group D and group E had better effect than group C .Conclusion Blueberry has a good protective effect on acute hepatic injury in rats .Its mechanism would be related to anti oxidative stress ,and partly through up‐regu‐lating the expression of GCLC .