小茴香精油对肝纤维化大鼠抗氧化应激损伤作用的研究

Antioxidant effects of the Uygur herb, Foenicuinm Vulgare Mill, in a rat model of hepatic fibrosis

目的评价小茴香精油在肝纤维化大鼠馍型中抗氧化应激损伤及抗肝纤维化的作用。方法肝纤维化模型的构建：SD大鼠皮下注射4ml／kg四氯化碳橄榄油溶液，同时给予高脂低蛋白饲料共5周。设空白对照组（A组）8只，正常饲养；预防阶段模型对照组（B组）10只，造模同时给予等渗盐水灌胃，小茴香预防组（C组）10只，造模同时给予小茴香精油灌胃，剂量为3ml／kg；治疗阶段模型对照组（D组）22只，造模后给予等渗盐水灌胃。小茴香治疗组（E组）22只，造模后给予小茴香精油灌胃，剂量为3ml／kg，扶正化瘀对照组（F组）22只，造模后给予扶正化瘀胶囊与等渗盐水悬混液灌胃，剂量为0．4g／kg。分别于第5周末处死A、B、C组大鼠，第6、7、8、9周末处死D、E、F组各4～6只大鼠。检测血清ALT、AST、透明质酸、层黏蛋白、8-羟基-2-脱氧鸟嘌呤含量，肝组织匀浆超氧化物歧化酶、谷胱甘肽过氧化物酶、丙二醛。肝组织行HE染色、马松染色和α平滑肌肌动蛋白免疫组织化学染色。计量资料先进行正态性检验和方差齐性检验，满足方差分析条件时采用两因素析因设计的方差分析，差异有统计学意义后两两比较采用LDL检验。不满足方差分析条件和等级资料采用秩和检验。结果A、B、C3组超氧化物歧化酶活力差异有统计学意义（286．33±37．90）U／mg、（242．22±33．21）U／mg、（323．78±53．16）U／mg，F=8．649，P=0．002），谷胱甘肽过氧化物酶分别为（297．25±54．29）U／mg、（148．62±15．57）U／mg、（221．65±53．39）U／mgF=26．468P=0．000）I丙二醛分别为（1．01±0．37）nmol／mg、（4．29±1．85）nmol／mg、（2．00±0．42）nmol／mgF=18．13，P=0．000），8-羟基-2-脱氧鸟嘌呤含量含量为（35．44±2．19）ng／L、（50．25±8．29）ng／L、（39．20±4．00）ng／LF=16．332，P=0．000）。炎症活动度在c组明显低于B组f=-2．29P=0．022）；E组轻于D组F=-2．51P=0．012），F组轻于D组F=-2．30，P=0．021），纤维化分期E组与D组差异有统计学意义F=-2．81，P=0．005），F组与D组比较差异有统计学意义F=-2．59P=0．015）。结论小茴香精油具有减轻肝脏炎症反应、防治肝纤维化进展的作用，其机制可能与抗氧化应激损伤有关。

Objective To evaluate therapeutic and antioxidant effects of Uygur Herb Foeniculum Vulgare Mill （FVM） in hepatic fibrosis rats. Method Hepatic fibrosis model was built in rats by subcutaneous injection with 40% CCl4 olive oil mixture. At the same time the rats were given high lipoid-low protein animal feeds for 5 weeks. 94 male SD rats were randomly divided into six groups ：blank control group （A-group）, 8 rats were feed in normal; prevention model control group （B-group）, 10 rats were given saline solution by intragastric administration during make of hepatic fibrosis model; FVM prevention group （C-group）, 10 rats were given FVM by intragastric administration during make of hepatic fibrosis model; model control group （D-group）, FVM treatment group （E-group）; Fuzhenghuayu treatment group （F-group）. 22 rats in each D, E, F-group were respectively given saline solution, FVM and Fuzhenghuayu by intragastric administration after hepatic fibrosis model were built. At the 5-th weekend, A, B, C- group rats were sacrificed. At the 6-th, 7-th, 8-th, 9-th weekend, 4-6 rats in D, E, F-group were sacrificed. Serum alanine aminotransferase （ALT）, aspartate aminotransferase （AST）, hyaluronic acid （HA）, laminin （LN） and 8 - hydroxy-2-deoxyguanosine （8- OHdG） were detected, liver tissue homogenate superoxide dismutase （SOD）, glutathione peroxidase （GSH- Px）, malondialdehyde （MDA） were detected. Histopathologic changes were observed after H.E and Masson staining. The expression of alpha-smooth muscle actin（α-SMA） were detected by immunohistochemical staining. The data were analyzed by SPSS17.0 software. Results The serum levels of ALT, AST, HA, and LN in the FVM prevention group were significantly reduced compared to the prevention model control group. （P 〈 0.05）. Rats in FVM treatment group appeared a marked lower serum levels of ALT, AST, HA compared to the model control group （P 〈 0.05）, and a distinguished lower Inflammation grade and fibrosis stage （P 〈 0.05） when the liver section were assayed as well; Rats in FVM treatment group and FVM prevention group had a conspicuous lower content of MDA, 8-OHdG, fibre and α-SMA expression （P 〈 0.05）, a significantly higher level of SOD, GSH-Px comared to those of in the model control groups. Conclusions Foeniculum Vulgare Mill declines liver inflammation response ,and prevent the hepatic fibrosis progression,, this may be due to its effects of antioxidative results.