加味揉肝汤提取物对雏鸡急性肝损伤的缓解作用

Extracts of Modified Rougan Decoction alleviated acute liver injury in chicken

制备加味揉肝汤水提物(MRGD)和水提沉淀物(多糖,MRGDE);1日龄的海兰雏鸡随机分为4组(n=16):对照组(Control)、脂多糖-恩诺沙星(LPS-ENR)组和药物组(MRGD+LPS-ENR、MRGDE+LPS-ENR),第4天药物组口服中药1h后,除对照组外其余鸡群口服恩诺沙星,连续3d。于第6天除对照组外其余各组腹腔注射LPS,造成雏鸡急性肝损伤。24h后采集血清和肝脏样品,检测血清肝生化指标;观察肝脏病理变化,肝脏匀浆液测定氧化应激评价指标的影响,免疫荧光法观察肝组织中Nrf2蛋白的表达,qPCR法检测抗氧化基因的mRNA水平,Western blot法检测Nrf2和CAT的蛋白表达的变化。研究结果显示,与LPS-ENR组相比较,药物MRGD和MRGDE的干预使雏鸡肝脏肿胀程度降低,肝脏组织结构被改善;血清AST、ALT水平下调,TP、ALB水平上调;肝脏抗氧化酶SOD和CAT活力升高,MDA水平降低;Nrf2、SOD、CAT和GPX的mRNA水平升高,Nrf2和CAT的蛋白表达水平升高;肝脏中Nrf2蛋白的荧光强度增加。MRGD和MRGDE通过激活Nrf2提高急性肝损伤雏鸡的抗氧化能力,抑制氧化应激水平,从而改善了LPS-ENR导致的肝脏损伤。

To explore the protective effect of Modified Rougan Decoction(MRGD) on acute liver injury in chickens.Water extracts(MRGD)and the alcohol precipitates of water extracts(polysaccharide,MRGDE)were prepared;three-day-old Hailan chicks were randomly divided into four groups(n=16):control group,lipopolysaccharide-enrofloxacin(LPS-ENR)group,and drug group(MRGD+LPS-ENR,MRGDE+LPS-ENR).On the 4th d,after 1hof oral administration of herbs,all chickens except for the control group received oral administration of enrofloxacin for 3d.On the 6th day,LPS was injected intraperitoneally,causing acute liver injury in the chicks.After 24h,serum and liver samples were collected to detect the influence of serum on biochemical indexes.The pathological changes in the liver were observed and the effect of liver homogenate on oxidative stress index was determined.The expression of the Nrf2protein in liver tissue was observed by immunofluorescence.The mRNA expression level of antioxidant genes was detected by the qPCR method.The protein expression of Nrf2and CAT was detected by Western blot.The results showed that compared with the LPS-ENR group,the degree of liver swelling in the MRGD and MRGDE groups decreased,and the liver tissue structure was improved.Serum biochemical indicators showed that AST and ALT levels were decreased,TP and ALB levels were increased;the activities of SOD and CAT in the liver increased,and the MDA level decreased;the mRNA levels of Nrf2,SOD,CAT and GPX increased,and the protein expression levels of Nrf2and CAT increased.The fluorescence intensity of the Nrf2protein in the liver increased.In conclusion,MRGD and MRGDE can improve the liver injury induced via activating the Nrf2pathway by improving the antioxidant capacity and inhibiting the level of oxidative stress in chickens with acute liver injury.