多烯磷脂酰胆碱对非酒精性脂肪肝小鼠的干预及其对肝锌指蛋白A20表达的影响
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摘要
目的研究多烯磷脂酰胆碱(PPC)对非酒精性脂肪肝(NAFLD)小鼠的干预作用及其对肝锌指蛋白A20表达的影响。方法从30只C57BL/6雄性小鼠中随机选取10只饲喂普通饲料(对照组),20只饲喂甲硫氨酸胆碱缺乏(MCD)饲料用于构建NAFLD模型,并随机分为模型组和实验组,每组10只。实验组灌胃PPC 0.1 m L·kg~(-1),对照组和模型组灌胃生理盐水10 m L·kg~(-1),每天1次,连续灌服6周。用苏木精-伊红染色(HE)观察小鼠肝组织病理变化,用蛋白质免疫印迹法检测小鼠肝组织锌指蛋白A20表达,用酶联免疫吸附(ELISA)法测定小鼠血清炎症因子水平,用自动生化仪测定小鼠脂代谢水平。结果模型组和实验组非酒精性脂肪肝活动度评分(NAS)总分分别为(4.72±0.75),(1.43±1.12)分,均明显高于对照组的0分(均P<0.01),且实验组低于模型组(P<0.01)。蛋白质免疫印迹法检测可见,锌指蛋白A20表达量从高到低依次为模型组、实验组、对照组。模型组小鼠血清肿瘤坏死因子-α(TNF-α)、单核细胞趋化蛋白-1(MCP-1)和白细胞介素-6(IL-6)水平均显著高于对照组(P<0.05,P<0.01)。与对照组比较,模型组小鼠血清三酰甘油及游离脂肪酸水平升高,总胆固醇水平降低(P<0.05或P<0.01);与模型组比较,实验组三酰甘油升高,总胆固醇水平降低(均P<0.05)。结论 PPC可有效降NAFLD模型小鼠血清炎症因子水平,改善肝组织形态及脂代谢水平,可能与降低肝锌指蛋白A20表达量,调控其所介导的核因子-κB(NF-κB)炎症反应通路有关。
Objective To study the effect of polyene phosphatidylcholine( PPC) on mice with nonalcoholic fatty liver disease( NAFLD) and its effect on hepatic zinc finger protein A20 expression. Methods Ten cases among 30 C57 BL/6 mice were randomly selected as the control group,which were fed with ordinary feed,20 mice were modeled NAFLD mice which were fed with methionine and cholinedeficiency( MCD) feed,and then were randomly divided into model group and test group,10 cases in each group. The test group was gavaged with PPC 0. 1 m L·kg~(-1),control group and model group were gavaged with normal saline 10 m L·kg~(-1),once a day,both were continuously gavaged for 6 weeks.The histopathological changes of liver tissue were observed by hematoxylin-eosin staining( HE),the expression of zinc finger protein A20 were detected by Western blotting,the levels of serum inflammatory cytokines were measured by enzyme-linked immunoso-rbent assay( ELISA),the levels of lipid metabolism indexes was determined by automatic biochemical analyzer. Results The non-alcoholic fatty liver disease activity scores( NAS)in model group and test group were 4. 72 ± 0. 75 and 1. 43 ± 1. 12,higher than that in control group,which was 0( all P < 0. 01),test group was lower than model group( P < 0. 01). Western blotting test showed that the expression of zinc finger protein A20 from high to low were model group,test group,control group. The levels of tumor necrosis factor-α( TNF-α),monocyte chemoattractant protein-1( MCP-1) and interleukin-6( IL-6) in model group were higher than those in control group( P < 0. 05 or P < 0. 01). Compared with control group,the levels of serum triglyceride and free fatty acid in model group increased,the total cholesterol level decreased( P < 0. 05 or P < 0. 01),compared with model group,the triglyceride increased and total cholesterol level decreased in test group( all P < 0. 05).Conclusion PPC can effectively reduce the levels of serum inflammatory factors in NAFLD model mice and improve the liver tissue morphology and lipid metabolism,which may be related to the decrease of hepatic zinc finger protein A20 expression and the regulation of nuclear factor-κB( NF-κB) inflammatory pathway.
Objective To study the effect of polyene phosphatidylcholine( PPC) on mice with nonalcoholic fatty liver disease( NAFLD) and its effect on hepatic zinc finger protein A20 expression. Methods Ten cases among 30 C57 BL/6 mice were randomly selected as the control group,which were fed with ordinary feed,20 mice were modeled NAFLD mice which were fed with methionine and cholinedeficiency( MCD) feed,and then were randomly divided into model group and test group,10 cases in each group. The test group was gavaged with PPC 0. 1 m L·kg~(-1),control group and model group were gavaged with normal saline 10 m L·kg~(-1),once a day,both were continuously gavaged for 6 weeks.The histopathological changes of liver tissue were observed by hematoxylin-eosin staining( HE),the expression of zinc finger protein A20 were detected by Western blotting,the levels of serum inflammatory cytokines were measured by enzyme-linked immunoso-rbent assay( ELISA),the levels of lipid metabolism indexes was determined by automatic biochemical analyzer. Results The non-alcoholic fatty liver disease activity scores( NAS)in model group and test group were 4. 72 ± 0. 75 and 1. 43 ± 1. 12,higher than that in control group,which was 0( all P < 0. 01),test group was lower than model group( P < 0. 01). Western blotting test showed that the expression of zinc finger protein A20 from high to low were model group,test group,control group. The levels of tumor necrosis factor-α( TNF-α),monocyte chemoattractant protein-1( MCP-1) and interleukin-6( IL-6) in model group were higher than those in control group( P < 0. 05 or P < 0. 01). Compared with control group,the levels of serum triglyceride and free fatty acid in model group increased,the total cholesterol level decreased( P < 0. 05 or P < 0. 01),compared with model group,the triglyceride increased and total cholesterol level decreased in test group( all P < 0. 05).Conclusion PPC can effectively reduce the levels of serum inflammatory factors in NAFLD model mice and improve the liver tissue morphology and lipid metabolism,which may be related to the decrease of hepatic zinc finger protein A20 expression and the regulation of nuclear factor-κB( NF-κB) inflammatory pathway.
引文
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[5]徐鑫,孟柠,俞毅君,等.大黄与甘草合用对肠功能衰竭大鼠炎性因子的影响研究[J].中国全科医学,2016,19(15):1824-1827.
[6]IMURA H,SHIMADA A,NAOTA M,et al.Vanadium toxicity in mice:possible impairment of lipid metabolism and mucosal epithelial cell necrosis in the small intestine[J].Toxicol Pathol,2013,41(6):842-856.
[7]刘浠,易威威,温亚枫,等.锌指蛋白A20及其相关炎症因子在椎间盘髓核细胞退变前后的表达变化[J].第三军医大学学报,2016,38(18):2077-2081.
[8]刘皎,杨洁,向颖,等.人原发性肝细胞癌中锌指蛋白A20、NF-κB p65及多药耐药P-gp蛋白的表达[J].中国普外基础与临床杂志,2012,19(12):1302-1306.
[9]ISHIZUKA M,OHTSUKA E,INOUE A,et al.Abnormal spermatogenesis and male infertility in testicular zinc finger protein Zfp318-knockout mice[J].Dev Growth Differ,2016,58(7):600-608.
[2]YOUNOSSI Z M,KOENIG A B,ABDELATIF D,et al.Global epidemiology of nonalcoholic fatty liver disease-Meta-analytic assessment of prevalence,incidence,and outcomes[J].Hepatology,2016,64(1):73-84.
[3]SAN-MIGUEL B,ALVAREZ M,CULEBRAS J M,et al.N-acetyl-cysteine protects liver from apoptotic death in an animal model of fulminant hepatic failure[J].Apoptosis,2006,11(11):1945-1957.
[4]TYAGI H,JHA S,SHARMA M,et al.Rice SAPs are responsive to multiple biotic stresses and overexpression of Os SAP1,an A20/AN1zinc-finger protein,enhances the basal resistance against pathogen infection in tobacco[J/OL].Trends Plant Sci,2014,225:68-76.2014-08-01[2017-05-22].https://www.sciencedirect.com/science/article/pii/S0168945214001277?via%3Dihub.
[5]徐鑫,孟柠,俞毅君,等.大黄与甘草合用对肠功能衰竭大鼠炎性因子的影响研究[J].中国全科医学,2016,19(15):1824-1827.
[6]IMURA H,SHIMADA A,NAOTA M,et al.Vanadium toxicity in mice:possible impairment of lipid metabolism and mucosal epithelial cell necrosis in the small intestine[J].Toxicol Pathol,2013,41(6):842-856.
[7]刘浠,易威威,温亚枫,等.锌指蛋白A20及其相关炎症因子在椎间盘髓核细胞退变前后的表达变化[J].第三军医大学学报,2016,38(18):2077-2081.
[8]刘皎,杨洁,向颖,等.人原发性肝细胞癌中锌指蛋白A20、NF-κB p65及多药耐药P-gp蛋白的表达[J].中国普外基础与临床杂志,2012,19(12):1302-1306.
[9]ISHIZUKA M,OHTSUKA E,INOUE A,et al.Abnormal spermatogenesis and male infertility in testicular zinc finger protein Zfp318-knockout mice[J].Dev Growth Differ,2016,58(7):600-608.
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