茵陈五苓散对胆汁淤积幼龄大鼠肝功能保护作用  

Effect and mechanism of Yinchen Wuling Powder on liver function in young rats with cholestasis

作　　者：王新芳 胡晨旻[1] 赵真 张志群[1] WANG Xin-fang;HU Chen-min;ZHAO Zhen(Pediatric Department,The First People's Hospital of Hangzhou City,Hangzhou,Zhejiang Province 310000,China)

机构地区：[1]杭州市第一人民医院儿科,浙江310000

出　　处：《中国公共卫生》2021年第10期1521-1526,共6页Chinese Journal of Public Health

基　　金：浙江省自然科学基金(LY15H040002)。

摘　　要：目的探讨茵陈五苓散对胆汁淤积幼龄大鼠肝功能的保护作用及可能机制。方法幼龄SD大鼠随机分为对照组、模型组、熊去氧胆酸组(UDCA, 0.10 g/kg)、茵陈五苓散高、中、低剂量组(100.0、200.0、400.0 g/kg),每组12只,雌雄各半。给药6 d后,采用0.5%α–萘异硫氰酸酯(ANIT, 50 mg/kg)花生油溶液灌胃,建立胆汁淤积模型。造模后继续给药2 d,最后一次给药12 h后,收集大鼠胆汁,生化法检测血清碱性磷酸酶(ALP)、γ–谷氨酰转移酶(γ-GT)、总胆汁酸(TBA)、丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AST)水平和肝匀浆中超氧化物歧化酶(SOD)、丙二醛(MDA)、过氧化氢酶(CAT)、谷胱甘肽过氧化物酶(GSH-Px)水平;苏木精–伊红染色(HE)观察各组大鼠肝组织病理变化;Western blot检测肝组织中Kelch样环氧氯丙烷相关蛋白1(Keap1)、核因子E2相关因子2(Nrf2)、还原型辅酶/醌氧化还原酶(NQO1)和血红素加氧酶1(HO-1)蛋白表达。结果与对照组比较,模型组大鼠血清ALP、γ-GT、TBA、ALT、AST水平均显著升高(P <0.05);胆汁流量、肝组织中SOD、CAT、GSH-Px水平显著降低,MDA水平明显增加(P <0.05);肝细胞索排列紊乱,细胞肿大伴点状坏死,有空泡样改变,大量炎性细胞浸润;肝组织Keap1蛋白表达显著升高,Nrf2、NQO1、HO-1蛋白表达显著降低(P <0.05)。与模型组比较,UDCA组、茵陈五苓散高、中剂量组大鼠血清ALP、γ-GT、TBA、ALT、AST水平均显著降低(P <0.05);胆汁流量、肝匀浆中SOD、CAT、GSH-Px水平均显著升高,MDA水平明显降低(P <0.05);肝组织病理损伤减轻,Keap1蛋白表达明显降低,Nrf2、NQO1、HO-1蛋白表达明显升高(P <0.05)。结论茵陈五苓散对ANIT诱导幼龄大鼠胆汁淤积型肝损伤具有保护作用,其机制可能与抑制Keap1表达,上调Nrf2表达,提高机体的抗氧化能力,减轻氧化应激损伤有关。Objective To examine protective effect and mechanism of Yinchen Wuling Powder(composed of Chinese medicinal herbs including Artemisia capillaris, Poria cocos, Rhizoma alismatis, Polyporus umbellatus, Ramulus cinnamomi,and Rhizoma atractylodis) on liver function in young rats with cholestasis. Methods Totally 72 young Sprague-Dawley(SD) rats were randomly divided into 6 groups(6 males and 6 females in each group): a blank control and a model group with normal saline, an ursodeoxycholic acid(UDCA, at the dose of 0.10 g/kg) group, and low, moderate, high dose(100,200, 400 g/kg) Yinchen Wuling Powder groups. The administrations were conducted by gavage once a day continuously for6 days before and 2 day after the establishment of cholestasis model induced by intragastric administration of 0.5%α-naphthalene isothiocyanate(ANIT) at the dosage of 50 mg/kg in all rats except for those of control group. Twelve hours after the last administration, all rats′ bile were collected and blood specimens were sampled for detections of serum alkaline phosphatase(ALP), γ-glutamyltransferase(γ-GT), total bile acid(TBA), alanine aminotransferase(ALT), and aspartate aminotransferase(AST) with biochemical method;the rats′ liver tissue specimens were also sampled for detections of superoxide dismutase(SOD), malondialdehyde(MDA), catalase(CAT), and glutathione peroxidase(GSH-Px) in liver homogenate with biochemical method, the observation of pathological changes in liver tissues with hematoxylin-eosin(HE)staining;and determinations of protein expression of Kelch-like ECH-associated protein 1(Keap1), nuclear factor erythroid-2-related factor-2(Nrf2), reduced coenzyme/quinone oxidoreductase(NQO1), and heme oxygenase 1(HO-1) with Western blot. Results Compared with those in the blank control rats, significantly higher serum ALP, γ-GT, TBA, ALT and AST and lower bile secretion, SOD, CAT and GSH-Px in liver tissues were detected in the model rats(P < 0.05 for all);disordered arrangement of hepatocyte cords, enlargement/spotty necrosis/v

关 键 词：茵陈五苓散 Kelch样环氧氯丙烷相关蛋白1/核因子E2相关因子2(Keap1/Nrf2) 胆汁淤积 肝损伤 幼龄大鼠 

分 类 号：R99[医药卫生—毒理学]