山楂酸对高脂饮食诱导的非酒精性脂肪肝模型小鼠炎症反应及氧化应激的影响
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摘要
目的:探讨山楂酸对高脂饮食诱导的非酒精性脂肪肝(NAFLD)模型小鼠炎症反应及氧化应激的影响。方法:将72只C57BL/6小鼠随机分为正常组(生理盐水)、模型组(生理盐水)、辛伐他汀组(阳性对照,3 mg/kg)和山楂酸低、中、高剂量组(25、50、100 mg/kg),每组12只。正常组小鼠给予标准饲料喂养,其余各组小鼠给予高脂饲料喂养以诱导NAFLD模型。造模同时,各组小鼠灌胃相应剂量的药物,每天给药1次,共计给药12周。末次给药后12 h,称量各组小鼠体质量及肝质量,并计算肝指数;全自动生化分析仪测定其血清生化指标[天冬氨酸转氨酶(AST)、丙氨酸转氨酶(ALT)活性及高密度脂蛋白胆固醇(HDL-C)、低密度脂蛋白胆固醇(LDL-C)、三酰甘油(TG)、总胆固醇(TC)含量]变化;苏木精-伊红染色后观察其肝组织病理形态变化;酶联免疫吸附试验法检测其肝组织中炎症反应指标[核转录因子κB(NF-κB)、肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)含量]和分光光度法测定其中氧化应激指标[丙二醛(MDA)含量及超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)活性]变化。结果:与正常组比较,模型组小鼠肝指数显著升高(P<0.05);血清中HDL-C含量显著降低,其余血清生化指标活性/含量均显著升高(P<0.05);肝小叶界限不清晰,肝组织发生明显病理性改变;肝组织中炎症反应指标和MDA含量均显著升高(P<0.05),SOD、GSH-Px活性显著降低(P<0.05)。与模型组比较,除山楂酸低剂量组小鼠肝组织中SOD活性升高和MDA含量降低不显著外(P>0.05),其余各给药组小鼠上述指标均显著改善(P<0.05);肝组织中球形脂滴及炎性细胞浸润减少。结论:山楂酸可通过抗炎及抑制氧化应激来减轻高脂饮食诱导的NAFLD模型小鼠的肝组织病变程度。
OBJECTIVE:To investigate the effects of crataegolic acid on inflammatory response and oxidative stress in non-alcoholic fatty liver disease(NAFLD)model mice induced by high-fat diet. METHODS:Totally 72 C57 BL/6 mice were randomly divided into normal group(normal saline),model group(normal saline),simvastatin group(positive control,3 mg/kg) and crataegolic acid low-dose,medium-dose and high-dose groups(25,50 and 100 mg/kg),with 12 mice in each group. Normal group was standard feed,while other groups were given high-fat diet to induce NAFLD model. At the same time of modeling,rats in each group was given relevant dose of drugs intragastrically, once a day, for consecutive 12 weeks. 12 h after last administration,body weight and liver weight of mice were determined,and liver index was calculated. The changes of serum biochemical indexes(activities of AST,ALT and contents of HDL-C,LDL-C,TG,TC) were determined by fully automatic biochemical analyzer. The pathological changes of liver tissue were observed after HE staining. The changes of inflammatory response indexes(contents of NF-κB,TNF-α,IL-6)in liver tissue were determined by ELISA,and the changes of oxidant stress indexes(contents of MDA,SOD and activity of GSH-Px) were determined by spectrophotometry. RESULTS:Compared with normal group,the liver index of mice in model group was increased significantly(P<0.05). The serum content of HDL-C was decreased significantly,and the activities/contents of other serum biochemical indexes were increased significantly(P<0.05). The boundary of hepatic lobules was not clear,and the liver tissue had obvious pathological changes. Inflammatory response indexes and the contents of MDA were increased significantly in liver tissue(P<0.05),the activities of SOD and GSH-Px were decreased significantly(P<0.05). Compared with model group,except that the increase of SOD activity and the decrease of MDA content of liver tissue were not significantly in crataegolic acid low-dose group(P>0.05), while above indexes of mice in other administration groups were improved significantly(P<0.05). The globular lipid droplets and inflammatory cell infiltration were decreased in liver tissue. CONCLUSIONS:Crataegolic acid can effectively alleviate the degree of liver lesions in NAFLD model mice induced by high-fat die through anti-inflammatory and inhibiting oxidative stress.
OBJECTIVE:To investigate the effects of crataegolic acid on inflammatory response and oxidative stress in non-alcoholic fatty liver disease(NAFLD)model mice induced by high-fat diet. METHODS:Totally 72 C57 BL/6 mice were randomly divided into normal group(normal saline),model group(normal saline),simvastatin group(positive control,3 mg/kg) and crataegolic acid low-dose,medium-dose and high-dose groups(25,50 and 100 mg/kg),with 12 mice in each group. Normal group was standard feed,while other groups were given high-fat diet to induce NAFLD model. At the same time of modeling,rats in each group was given relevant dose of drugs intragastrically, once a day, for consecutive 12 weeks. 12 h after last administration,body weight and liver weight of mice were determined,and liver index was calculated. The changes of serum biochemical indexes(activities of AST,ALT and contents of HDL-C,LDL-C,TG,TC) were determined by fully automatic biochemical analyzer. The pathological changes of liver tissue were observed after HE staining. The changes of inflammatory response indexes(contents of NF-κB,TNF-α,IL-6)in liver tissue were determined by ELISA,and the changes of oxidant stress indexes(contents of MDA,SOD and activity of GSH-Px) were determined by spectrophotometry. RESULTS:Compared with normal group,the liver index of mice in model group was increased significantly(P<0.05). The serum content of HDL-C was decreased significantly,and the activities/contents of other serum biochemical indexes were increased significantly(P<0.05). The boundary of hepatic lobules was not clear,and the liver tissue had obvious pathological changes. Inflammatory response indexes and the contents of MDA were increased significantly in liver tissue(P<0.05),the activities of SOD and GSH-Px were decreased significantly(P<0.05). Compared with model group,except that the increase of SOD activity and the decrease of MDA content of liver tissue were not significantly in crataegolic acid low-dose group(P>0.05), while above indexes of mice in other administration groups were improved significantly(P<0.05). The globular lipid droplets and inflammatory cell infiltration were decreased in liver tissue. CONCLUSIONS:Crataegolic acid can effectively alleviate the degree of liver lesions in NAFLD model mice induced by high-fat die through anti-inflammatory and inhibiting oxidative stress.
引文
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[15]EBRAHIMI-MAMEGHANI M,JAMALI H,MAHDAVIR,et al.Conjugated linoleic acid improves glycemic response,lipid profile,and oxidative stress in obese patients with non-alcoholic fatty liver disease:a randomized controlled clinical trial[J].Croat Med J,2016,57(4):331-342.
[16]钱秋海,钱卫斌,蔡欣蕊,等.糖肝康对糖尿病非酒精性脂肪肝模型大鼠血清GSH、GSH-Px表达的影响[J].中医学报,2016,31(4):499-501,505.
[17]SUNDARAM SS,HALBOWER A,PAN Z,et al.Nocturnal hypoxia-induced oxidative stress promotes progression of pediatric non-alcoholic fatty liver disease[J].JHepatol,2016,65(3):560-569.
[18]K?RO?LU E,CANBAKAN B,ATAY K,et al.Role of oxidative stress and insulin resistance in disease severity of non-alcoholic fatty liver disease[J].Turk J Gastroenterol,2016,27(4):361-366.
[2]ALBHAISI S,ISSA D,ALKHOURI N.Non-alcoholic fatty liver disease:a pandemic disease with multisystem burden[J].Hepatobiliary Surg Nutr,2018,7(5):389-391.
[3]刁琢,冯旭,储智勇,等.山楂酸的药理作用及结构修饰研究进展[J].中国食品添加剂,2016,21(8):210-218.
[4]王颖,蔡永青,黄明春,等.山楂酸对四氯化碳致小鼠急性肝损伤的保护作用及机制研究[J].中国临床药理学与治疗学,2016,21(8):854-858.
[5]JAVADI L,KHOSHBATEN M,SAFAIYAN A,et al.Proand prebiotic effects on oxidative stress and inflammatory markers in non-alcoholic fatty liver disease[J].Asia Pac JClin Nutr,2018,27(5):1031-1039.
[6]FUJII J,HOMMA T,KOBAYASHI S,et al.Mutual interaction between oxidative stress and endoplasmic reticulum stress in the pathogenesis of diseases specifically focusing on non-alcoholic fatty liver disease[J].World J Biol Chem,2018,9(1):1-15.
[7]FU D,CUI H,ZHANG Y.Lack of ClC-2 alleviates high fat diet-induced insulin resistance and non-alcoholic fatty liver disease[J].Cell Physiol Biochem,2018,45(6):2187-2198.
[8]符恒.辛伐他汀治疗非酒精性脂肪性肝炎的疗效观察[J].内科,2014,9(6):677-678.
[9]谭丹枫,陶月英,姜媛,等.胆石六号颗粒对模型小鼠非酒精性脂肪肝的改善作用研究[J].中国药房,2018,29(22):3106-3110.
[10]LEI L,ZHOU C,YANG X,et al.Down-regulation of microRNA-375 regulates adipokines and inhibits inflammatory cytokines by targeting AdipoR2 in non-alcoholic fatty liver disease[J].Clin Exp Pharmacol Physiol,2018,45(8):819-831.
[11]ASSUN??O SNF,SORTE NCAB,ALVES CAD,et al.Inflammatory cytokines and non-alcoholic fatty liver disease(NAFLD)in obese children and adolescents[J].Nutr Hosp,2018,35(1):78-83.
[12]姚笑睿,夏凡,唐外姣,等.护肝清脂片对非酒精性脂肪肝大鼠肝脏中AMPK通路激活及NF-κB-p65蛋白的影响[J].南方医科大学学报,2017,37(1):56-62.
[13]唐彬,宋振梅,肖琦凡.黄芩苷对高脂-脂肪肝模型大鼠肝脏炎症改善及炎性因子表达的影响[J].青岛大学医学院学报,2017,53(6):659-663,667.
[14]ZHU CG,LIU YX,WANG H,et al.Active form of vitamin D ameliorates non-alcoholic fatty liver disease by alleviating oxidative stress in a high-fat diet rat model[J].Endocr J,2017,64(7):663-673.
[15]EBRAHIMI-MAMEGHANI M,JAMALI H,MAHDAVIR,et al.Conjugated linoleic acid improves glycemic response,lipid profile,and oxidative stress in obese patients with non-alcoholic fatty liver disease:a randomized controlled clinical trial[J].Croat Med J,2016,57(4):331-342.
[16]钱秋海,钱卫斌,蔡欣蕊,等.糖肝康对糖尿病非酒精性脂肪肝模型大鼠血清GSH、GSH-Px表达的影响[J].中医学报,2016,31(4):499-501,505.
[17]SUNDARAM SS,HALBOWER A,PAN Z,et al.Nocturnal hypoxia-induced oxidative stress promotes progression of pediatric non-alcoholic fatty liver disease[J].JHepatol,2016,65(3):560-569.
[18]K?RO?LU E,CANBAKAN B,ATAY K,et al.Role of oxidative stress and insulin resistance in disease severity of non-alcoholic fatty liver disease[J].Turk J Gastroenterol,2016,27(4):361-366.