自拟通络祛浊方对两种不同方法诱导大鼠非酒精性单纯性脂肪肝模型的干预效果及机制研究
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摘要
目的研究通络祛浊方对化学性肝损伤或高脂饲料诱导的大鼠非酒精性单纯性脂肪肝(NAFL)模型的干预效果,并探索作用机制。方法四氯化碳(tetrachloride,CCl4)造模实验设置溶媒组、CCl4模型组和通络祛浊方预防组(简称预防组);高脂饲料造模实验设置空白对照组、高脂饲料模型组和预防组。测定血生化、肝脏脂质含量、细胞色素P450主要亚酶活性,进行组织病理学观察及代谢组学分析等。结果通络祛浊方明显降低两种不同方法诱导的大鼠NAFL模型的肝脏脂质含量,降低肝脏质量及肝脏系数,对血中的甘油三酯(triglyceride,TG)有双向调节作用。CYP酶活性在CCl4模型组有明显降低,并在预防组有明显恢复。通络去浊方干预CCl4诱导的模型主要通过磷脂类物质代谢通道,而高脂饲料模型引起变化的主要代谢物质为游离脂肪酸类。结论通络祛浊方对两种不同因素诱导的大鼠NAFL模型具有较明显的保肝降脂的干预效果,其在不同方法诱导的NAFL中发挥干预作用的主要代谢产物及代谢通路不同。
Aim To investigate the effect of Tongluoquzhuo prescription on the intervention of non-alcoholic simple fatty liver( NAFL) rats established by chemical injury or a high-fat diet. Methods Three testing groups,vehicle group,NAFL group and intervention group were set in the experiment of NAFL rats established by carbon tetrachloride( CCl_4); and those of control group, NAFL group and intervention group were set in the experiment of NAFL rats established by a high-fat diet. Blood biochemical determination,contents of hepatic lipids, CYP enzyme activity, histopathological observation,and metabolomics analysis were measured. Results Tongluoquzhuo prescription reduced the hepatic lipids of NAFL rats established by different methods significantly, lowered liver weight and liver index,and had dual-direction regulation of blood triglyceride( TG). CYP enzyme activity was reduced significantly in NAFL rats established by CCl_4,and raised significantly in intervention group. It may be effected by the pathway of phospholipid to reduce lipid accumulation in the liver of NAFL rats established by CCl_4,while free fatty acid was the most important factor of the interventional effect in NAFL rats established by a high-fat diet. Conclusions Tongluoquzhuo prescription has evident effect of protecting liver and reducing lipids in NAFL rats established by different methods. On the other aspect,It shows difference between the main metabolites and metabolic pathways involved in the intervention of Tongluo turbidity in NAFL established by different methods. It is suggested that different metabolites are required as biomarkers,although different causes can produce similar hepatic steatosis.
Aim To investigate the effect of Tongluoquzhuo prescription on the intervention of non-alcoholic simple fatty liver( NAFL) rats established by chemical injury or a high-fat diet. Methods Three testing groups,vehicle group,NAFL group and intervention group were set in the experiment of NAFL rats established by carbon tetrachloride( CCl_4); and those of control group, NAFL group and intervention group were set in the experiment of NAFL rats established by a high-fat diet. Blood biochemical determination,contents of hepatic lipids, CYP enzyme activity, histopathological observation,and metabolomics analysis were measured. Results Tongluoquzhuo prescription reduced the hepatic lipids of NAFL rats established by different methods significantly, lowered liver weight and liver index,and had dual-direction regulation of blood triglyceride( TG). CYP enzyme activity was reduced significantly in NAFL rats established by CCl_4,and raised significantly in intervention group. It may be effected by the pathway of phospholipid to reduce lipid accumulation in the liver of NAFL rats established by CCl_4,while free fatty acid was the most important factor of the interventional effect in NAFL rats established by a high-fat diet. Conclusions Tongluoquzhuo prescription has evident effect of protecting liver and reducing lipids in NAFL rats established by different methods. On the other aspect,It shows difference between the main metabolites and metabolic pathways involved in the intervention of Tongluo turbidity in NAFL established by different methods. It is suggested that different metabolites are required as biomarkers,although different causes can produce similar hepatic steatosis.
引文
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[2]Ashtari S,Pourhoseingholi M A,Zali M R.Nonalcoholic fatty liver disease in Asia:prevention and planning[J].World J Hepatol,2015,7(13):1788-96.
[3]陈一平,肖百泉,张景鸿,等.新型非酒精性脂肪肝模型建立及免疫细胞机制探讨[J].中国药理学通报,2018,34(6):882-6.[3]Chen Y P,Xiao B Q,Zhang J H,et al.New method of non-alcoholic fatty liver disease mouse models and mechanism about immune cells[J].Chin Pharmacol Bull,2018,34(6):882-6.
[4]董丽红,张瑞芬,黄菲,等.油酸诱导单纯性脂肪肝变性细胞模型的建立及应用[J].中国药理学通报,2017,33(11):1662-6.[4]Dong L H,Zhang R F,Huang F,et al.Establishment and application of hepatocyte steatosis models induced by oleic acid[J].Chin Pharmacol Bull,2017,33(11):1662-6.
[5]魏华凤,季光.中药口服治疗非酒精性脂肪肝临床随机对照试验的系统评价[J].中华中医药杂志,2012,27(5):1309-14.[5]Wei H F,Ji G.Systematic review of treating nonalcoholic fatty liver with taking TCM orally:a clinical randomized controlled trial[J].Chin J Tradit Chin Med Pharm,2012,27(5):1309-14.
[6]贺建国.调脂复肝汤治疗非酒精性脂肪肝60例[J].中国医药科技,2013,20(6):697.[6]He J G.Observation on 60 cases of nonalcoholic fatty liver by decoction for relieving fat and recovering the liver[J].Chin J Tradit Med Sci Tec,2013,20(6):697.
[7]金毅,邢伟,吕爱贞,等.不同方法诱导大鼠非酒精性单纯性脂肪肝模型的CYP酶活性比较研究[C].第十四届中国实验动物科学年会论文集(光盘),2018:43-51.[7]Jin Y,Xing W,Lyu A Z,et al.Comparison of CYP enzyme activities in rat nonalcoholic fatty liver models established by different methods[C].The 14th academic conference on laboratory animal science of China(CD),2018:43-51.
[8]金毅,宋向荣,付新录,等.毒性病理学实用方法与技术[M].南京:江苏凤凰科学技术出版社,2017:318-50.[8]Jin Y,Song X R,Fu X L,et al.Practical methods and techniques of toxicological pathology[M].NanJing:Jiangsu Phoenix Science and Technology Press,2017:318-50.
[9]金毅,黄宝明,吕爱贞,等.四氯化碳诱导大鼠非酒精性脂肪肝的生物标记物研究[J].中国药师,2018,21(4):570-6.[9]Jin Y,Huang B M,Lyu A Z,et al.Study on the biomarkers of CCl4-induced non-alcoholic fatty liver disease in rats[J].Chin Pharmacist,2018,21(4):570-6.
[10]Chetwynd A J,Abdulsada A,Holt S G,et al.Use of a pre-analysis osmolality normalisation method to correct for variable urine concentrations and for improved metabolomic analyses[J].J Chromatogr A,2016,1431:103-10.
[11]Farrell G C,Chitturi S,Lau G K,et al.Guidelines for the assessment and management of non-alcoholic fatty liver disease in the A-sia-Pacific region:executive summary[J].J Gastroenterol Hepatol,2007,22(6):775-7.
[12]晨星燃,卞勉励,张晨曦,等.白介素1家族在脂肪肝疾病中的作用及机制研究进展[J].中国药理学通报,2017,33(6):753-6.[12]Chen X R,Bian M L,Zhang C X,et al.Research progress on role and mechanism of interleukin-1 in fatty liver diseases[J].Chin Pharmacol Bull,2017,33(6):753-6.
[13]Wang Y,Niu M,Jia G L,et al.Untargeted metabolomics reveals intervention effects of total turmeric extract in a rat model of nonalcoholic fatty liver disease[J].Evid Based Complement Alternat Med,2016,2016(4):1-12.