姜黄素对非酒精性脂肪性肝炎的氧化和抗氧化调节作用

The regulatory effects of curcumin on oxidation and antioxidation in non-alcoholicsteatohepatitis

目的 研究姜黄素对非酒精性脂肪性肝炎（non-alcoholic steatohepatitis,NASH）氧化和抗氧化调节作用.方法 清洁级雄性SD大鼠56只,使用随机数字表随机分7组.正常对照组8只,给予普通饲料喂养12周;模型组24只,给予胆碱缺乏（choline deficient,CD）饮食喂养,分别在4周、8周和12周随机处死8只;姜黄素治疗组24只,分为低剂量组（8只）、中剂量组（8只）、高剂量组（8只）,在CD饮食诱导4周之后分别添加姜黄素50、100和500 mg/（kg·d）喂养8周,12周时处死.留取肝脏组织进行病理学检测和硫代巴比妥酸反应物（thiobarbituric acid reactive substances,TBARS）、还原型谷胱甘肽（glutathione,GSH）、超氧化物歧化酶（superoxide dismutase,SOD）、锰超氧化物歧化酶（manganese superoxide dismutase,MnSOD）、谷胱甘肽过氧化物酶（glutathione peroxidase activity,GPx）、三酰甘油（triglyceride,TG）和总胆固醇（total cholesterol,TC）检测.符合正态分布的计量资料采用t检验,非正态分布资料采用秩和检验.结果 CD饮食诱导4周大鼠肝脏即具有NASH表现,8周时出现早期纤维化,12周时部分鼠进展为肝硬化.中、高剂量治疗组NAS评分为6.50（5.25,7.00）和6.00（5.00,6.75）,均低于12周模型组的8.00（7.00,8.00）,比较差异均有统计学意义（Z值分别为2.441和2.728,均P〈0.01）,但纤维化分期与12周模型组比较差异均无统计学意义（Z值分别为0.795和1.807,均P〉0.05）.低、中、高剂量治疗组大鼠肝脏TG（t值分别为0.54、1.18和1.66）和TC含量（t值分别为0.11、0.59和0.62）与12周模型组差异均无统计学意义（均P〉0.05）.高剂量治疗组大鼠肝脏GSH含量为（1185.82±204.01）mg/g,与12周模型组（735.29±35.08）mg/g比较差异有统计学意义（t=4.97,P〈0.01）;中、高剂量组大鼠肝脏TBARS含量与12周模型组比较差异均有统计学意义（t值分别为7.58和11.62,均P〈0.01）.低、中、高剂量组肝脏SOD含量与12周模型组比较,差异均有统计学意义（t值分别为4.17、4.32和6.10,均P〈0.01）;高剂量组肝脏M nSOD含量与12周模型组比较差异有统计学意义（t=8.42,P〈0.01）;而低、中、高剂量组肝脏GPx含量与12周模型组比较差异均无统计学意义（t值分别为0.27、0.21和0.60,均P〉0.05）.结论 CD饮食诱导SD大鼠时能引起肝脏脂质沉积,引起肝脏抗氧化系统障碍,GSH耗竭,SOD和GPx活力下降.姜黄素可改善NASH大鼠肝脏NAS评分,且可能通过上调机体SOD活力、提高肝脏GSH含量,发挥保护作用;而姜黄素对NASH大鼠肝脏GPx活力和脂质沉积无影响.

Objective To explore the regulatory effects of cucumin onoxidation and antioxidation in non-alcoholic steatohepatitis （NASH ） .Methods Fifty-six clean male rats were randomly divided into 7 groups by random numbers table .Eight rats in normal control group were fed by normal diet for 12 weeks .Twenty-four rats in model group were fed by choline dificinet （CD） diet and randomly sacrificedat week 4 ,8 and 12 with 8 rats each time point .Twenty-four rats incucumin treatment group were given cucumin at high （500 mg/[kg · d]） ,medium （100 mg/[kg · d]） and low （50 mg/[kg · d]） dosages with 8 rats each dosage from week 5 of CD diet for 8 weeks ,and the rats were sacrificed at week 12 .The liver tissues were reserved for pathology test and detections of thiobarbituric acid reactive substances （TBARS） ,glutathione （GSH） ,the activities of superoxide dismutase （SOD） ,manganese superoxide dismutase （MnSOD） ,glutathione peroxidase （GPx） ,and levels of triglyceride （TG） and total cholesterol （TC） .The measurement data with normal distribution were analyzed using t test ,and the data with non-normal distribution were analyzed using rank sum Z test .Results The liver of rats presented with the performance of NASH when fed with CD diet for 4 weeks ,and presented with early fibrosis after 8 weeks of CD diet ,even progressed to cirrhosis after 12 weeks of CD diet .The NAS scores of medium and high dose curcumin treatment groups were 6 .50 （5 .25 ,7 .00） and 6 .00 （5 .00 ,6 .75） ,respectively ,which were both lower than that model group at week 12 （8 .00 [7 .00 ,8 .00]）（Z=2 .441 and 2 .728 ,respectively , both P〈 0 .01） ,while fibrosis stages at week 12 were not significantly different compared with model group （Z=0 .795 and 1 .807 ,respectively ,both P〉 0 .05） .TG and TC levels in liver tissues of rats in low ,medium and high doses treatment group were not significantly different compared with model group at week 12（TG ：t=0 .54 ,1 .18 and 1 .66 ,respectively ;TC ：t=0 .11 ,0 .59 and 0 .62 ,respectively ;all P〉0 .05） .The GSH contentin liver of rats in high dose group was （1185 .82＋204 .01） mg/g ,which was significantly different from that in model group at week 12 （735 .29 ＋ 35 .08） （t=4 .97 ,P〈0 .01） .The TBARS contents in the liver of the middle and high doses curcumin treatment group were significantly different from that of model group at week 12 （t=7 .58 and 11 .62 ,respectively ,both P〈 0 .01） .The SOD activities in liver of rats in low ,medium and high doses curcumin treatment group were statistically different from that in model group （t=4 .17 ,4 .32 and 6 .10 ,respectively ,all P〈0 .01） .MnSOD activity in liver of rats in high dose group was significantly different from model group at week 12 （t=8 .42 ,P〈0 .01） .The live GPx contents in low ,medium and high doses curcumin treatment group were all not significantly different from that in model group at week 12 （t=0 .27 ,0 .21 and 0 .60 ,respectively ,all P〉0 .05） .Conclusions CD diet in SD rats could induce hepatic lipid deposition in liver ,and cause liver antioxidative system disorders ,GSH exhaustion ,and decreases of SOD and GPx activities .Curcumin treatment could improve liver NAS score of NASH rats ,and might play a protective role by upregulating the SOD activity and increasing liver GSH content .But curcumin has no effects on liver GPx activity and fat deposition in liver of NASH rats .