芪术方对高脂高果糖高胆固醇诱导的非酒精性脂肪性肝病小鼠模型的影响及其机制
目的 探讨芪术方对非酒精性脂肪性肝病 (NAFLD) 小鼠模型的疗效及作用机制。 方法 60只雄性C57BL/6J小鼠随机分为正常组，模型组，芪术方低 (4.75 g/kg) 、中 (9.50 g/kg) 、高剂量组 (19.00 g/kg) ，多唏磷脂酰胆碱组 (磷脂组) (228 mg/kg) ，每组10只。高脂高胆固醇饲料和20%果糖水造模16周后，各组给予相应药物干预，每天给药1次，连续8周。检测血清ALT、AST、TC、TG、LDL；酶联免疫吸附法 (ELISA) 检测血清游离脂肪酸 (FFA) 、肿瘤坏死因子-α (TNF-α) 、白细胞介素-1β (IL-1β) 、超氧化物歧化酶...
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| Published in | Linchuang gandanbing zazhi Vol. 40; no. 11; pp. 2205 - 2212 |
|---|---|
| Main Authors | , |
| Format | Journal Article |
| Language | Chinese |
| Published |
Changchun
Journal of Clinical Hepatology
01.11.2024
上海中医药大学附属曙光医院 细胞免疫实验室,上海 201203%上海中医药大学附属曙光医院肝病科,上海 201203 上海中医药大学附属曙光医院安徽医院肝病科,合肥 230011 |
| Subjects | |
| Online Access | Get full text |
| ISSN | 1001-5256 2097-3497 |
| DOI | 10.12449/JCH241113 |
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| Abstract | 目的 探讨芪术方对非酒精性脂肪性肝病 (NAFLD) 小鼠模型的疗效及作用机制。 方法 60只雄性C57BL/6J小鼠随机分为正常组，模型组，芪术方低 (4.75 g/kg) 、中 (9.50 g/kg) 、高剂量组 (19.00 g/kg) ，多唏磷脂酰胆碱组 (磷脂组) (228 mg/kg) ，每组10只。高脂高胆固醇饲料和20%果糖水造模16周后，各组给予相应药物干预，每天给药1次，连续8周。检测血清ALT、AST、TC、TG、LDL；酶联免疫吸附法 (ELISA) 检测血清游离脂肪酸 (FFA) 、肿瘤坏死因子-α (TNF-α) 、白细胞介素-1β (IL-1β) 、超氧化物歧化酶 (SOD) 、丙二醛 (MDA) ；苏木素-伊红 (HE) 和油红O染色观察肝组织病理变化；蛋白免疫印迹法 (Western Blot) 检测微管相关蛋白轻链3B (LC3BⅡ/Ⅰ) 、螯合体 (p62/SQSTM1) 、Beclin-1、动力相关蛋白1 (Drp1) 表达。实时荧光定量PCR (Real-time PCR) 检测Drp1、Beclin-1、p62/SQSTM1 mRNA表达水平。计量资料多组间比较采用单因素方差分析，进一步两两比较采用LSD-t检验。 结果 与正常组比较，模型组血清TG、TC、ALT、AST、LDL、FFA、TNF-α、IL-1β、MDA水平均升高，SOD水平降低 (P值均<0.05) 。HE染色结果显示，模型组小鼠肝组织可见肝细胞脂肪变及大量脂肪空泡；油红O染色结果显示，模型组小鼠肝细胞内有大量大小不一红色脂滴沉积，油红O染色面积百分比较正常组升高 (P<0.05) 。Real-time PCR结果显示，与正常组比较，模型组肝组织中Drp1、Beclin-1 mRNA升高，p62/SQSTM1 mRNA降低 (P值均<0.05) 。Western Blot结果显示，与正常组比较，模型组肝组织中Drp1、Beclin-1、LC3BⅡ/Ⅰ蛋白表达水平升高，p62/SQSTM1蛋白表达水平降低 (P值均<0.05) 。与模型组比较，部分剂量芪术方组和磷脂组血清TG、TC、ALT、AST、LDL、FFA、TNF-α、IL-1β、MDA水平均降低，SOD水平升高 (P值均<0.05) 。与模型组比较，各用药组肝组织脂肪变改善明显，且油红O染色面积百分比降低 (P值均<0.05) ；Drp |
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| AbstractList | 目的 探讨芪术方对非酒精性脂肪性肝病(NAFLD)小鼠模型的疗效及作用机制.方法 60只雄性C57BL/6J小鼠随机分为正常组,模型组,芪术方低(4.75 g/kg)、中(9.50 g/kg)、高剂量组(19.00 g/kg),多唏磷脂酰胆碱组(磷脂组)(228 mg/kg),每组10只.高脂高胆固醇饲料和20%果糖水造模16周后,各组给予相应药物干预,每天给药1次,连续8周.检测血清ALT、AST、TC、TG、LDL;酶联免疫吸附法(ELISA)检测血清游离脂肪酸(FFA)、肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、超氧化物歧化酶(SOD)、丙二醛(MDA);苏木素-伊红(HE)和油红O染色观察肝组织病理变化;蛋白免疫印迹法(Western Blot)检测微管相关蛋白轻链3B(LC3BⅡ/Ⅰ)、螯合体(p62/SQSTM1)、Beclin-1、动力相关蛋白1(Drp1)表达.实时荧光定量PCR(Real-time PCR)检测Drp1、Beclin-1、p62/SQSTM1 mRNA表达水平.计量资料多组间比较采用单因素方差分析,进一步两两比较采用LSD-t检验.结果 与正常组比较,模型组血清TG、TC、ALT、AST、LDL、FFA、TNF-α、IL-1β、MDA水平均升高,SOD水平降低(P值均<0.05).HE染色结果显示,模型组小鼠肝组织可见肝细胞脂肪变及大量脂肪空泡;油红O染色结果显示,模型组小鼠肝细胞内有大量大小不一红色脂滴沉积,油红O染色面积百分比较正常组升高(P<0.05).Real-time PCR结果显示,与正常组比较,模型组肝组织中Drp1、Beclin-1 mRNA升高,p62/SQSTM1 mRNA降低(P值均<0.05).Western Blot结果显示,与正常组比较,模型组肝组织中Drp1、Beclin-1、LC3BⅡ/Ⅰ蛋白表达水平升高,p62/SQSTM1蛋白表达水平降低(P值均<0.05).与模型组比较,部分剂量芪术方组和磷脂组血清TG、TC、ALT、AST、LDL、FFA、TNF-α、IL-1β、MDA水平均降低,SOD水平升高(P值均<0.05).与模型组比较,各用药组肝组织脂肪变改善明显,且油红O染色面积百分比降低(P值均<0.05);Drp1、Beclin-1 mRNA降低,p62/SQSTM1 mRNA升高(P值均<0.05);Drp1、Bec 目的 探讨芪术方对非酒精性脂肪性肝病 (NAFLD) 小鼠模型的疗效及作用机制。 方法 60只雄性C57BL/6J小鼠随机分为正常组，模型组，芪术方低 (4.75 g/kg) 、中 (9.50 g/kg) 、高剂量组 (19.00 g/kg) ，多唏磷脂酰胆碱组 (磷脂组) (228 mg/kg) ，每组10只。高脂高胆固醇饲料和20%果糖水造模16周后，各组给予相应药物干预，每天给药1次，连续8周。检测血清ALT、AST、TC、TG、LDL；酶联免疫吸附法 (ELISA) 检测血清游离脂肪酸 (FFA) 、肿瘤坏死因子-α (TNF-α) 、白细胞介素-1β (IL-1β) 、超氧化物歧化酶 (SOD) 、丙二醛 (MDA) ；苏木素-伊红 (HE) 和油红O染色观察肝组织病理变化；蛋白免疫印迹法 (Western Blot) 检测微管相关蛋白轻链3B (LC3BⅡ/Ⅰ) 、螯合体 (p62/SQSTM1) 、Beclin-1、动力相关蛋白1 (Drp1) 表达。实时荧光定量PCR (Real-time PCR) 检测Drp1、Beclin-1、p62/SQSTM1 mRNA表达水平。计量资料多组间比较采用单因素方差分析，进一步两两比较采用LSD-t检验。 结果 与正常组比较，模型组血清TG、TC、ALT、AST、LDL、FFA、TNF-α、IL-1β、MDA水平均升高，SOD水平降低 (P值均<0.05) 。HE染色结果显示，模型组小鼠肝组织可见肝细胞脂肪变及大量脂肪空泡；油红O染色结果显示，模型组小鼠肝细胞内有大量大小不一红色脂滴沉积，油红O染色面积百分比较正常组升高 (P<0.05) 。Real-time PCR结果显示，与正常组比较，模型组肝组织中Drp1、Beclin-1 mRNA升高，p62/SQSTM1 mRNA降低 (P值均<0.05) 。Western Blot结果显示，与正常组比较，模型组肝组织中Drp1、Beclin-1、LC3BⅡ/Ⅰ蛋白表达水平升高，p62/SQSTM1蛋白表达水平降低 (P值均<0.05) 。与模型组比较，部分剂量芪术方组和磷脂组血清TG、TC、ALT、AST、LDL、FFA、TNF-α、IL-1β、MDA水平均降低，SOD水平升高 (P值均<0.05) 。与模型组比较，各用药组肝组织脂肪变改善明显，且油红O染色面积百分比降低 (P值均<0.05) ；Drp |
| Abstract_FL | Objective To investigate the therapeutic effect and mechanism of action of Qizhu prescription in mice with non-alcoholic fatty liver disease(NAFLD).Methods A total of 60 male C57BL/6J mice were randomly divided into normal group,model group,low-dose Qizhu prescription group(4.75 g/kg),middle-dose Qizhu prescription group(9.50 g/kg),high-dose Qizhu prescription group(19.00 g/kg),Yishanfu group(228 mg/kg),with 10 mice in each group.After 16 weeks of modeling with a high-fat high-cholesterol diet and 20%fructose water,each group was given the corresponding drug once a day for 8 weeks.The serum levels of alanine aminotransferase(ALT),aspartate aminotransferase(AST),total cholesterol(TC),triglyceride(TG),and low-density lipoprotein(LDL)were measured;ELISA was used to measure the serum levels of free fatty acid(FFA),tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),superoxide dismutase(SOD),and malondialdehyde(MDA);HE staining and oil red O staining were used to the pathological changes of liver tissue;Western blot was used to measure the protein expression levels of LC3BⅡ/Ⅰ,p62/SQSTM1,Beclin-1,and Drp1,and real-time PCR was used to measure the mRNA expression levels of Drp1,Beclin-1,and p62/SQSTM1.A one-way analysis of variance was used for comparison of continuous data between multiple groups,and the least significant difference t-test was used for further comparison between two groups.Results Compared with the normal group,the model groups had significant increases in the serum levels of TG,TC,ALT,AST,LDL,FFA,TNF-α,IL-1β,and MDA and a significant reduction in the serum level of SOD(P<0.05).HE staining showed that the mice in the model group had hepatocyte steatosis and a large number of fat vacuoles in liver tissue,and oil red O staining showed that the mice in the model group had a large number of red lipid droplets of varying sizes in hepatocytes,with a significant increase in the percentage of oil red O staining area compared with the normal group(P<0.05).Real-time PCR showed that compared with the normal group,the model group had significant increases in the mRNA expression levels of Drp1 and Beclin-1 and a significant reduction in the mRNA expression level of p62/SQSTM1 in liver tissue(all P<0.05),and Western blot showed that compared with the normal group,the model group had significant increases in the protein expression levels of Drp1,Beclin-1,and LC3BⅡ/Ⅰ and a significant reduction in the protein expression level of p62/SQSTM1 in liver tissue(all P<0.05).Compared with the model group,some Qizhu prescription groups and the Yishanfu group had significant reductions in the serum levels of TG,TC,ALT,AST,LDL,FFA,TNF-α,IL-1β,and MDA and a significant increase in the serum level of SOD(all P<0.05).Compared with the model group,each administration group had a significant improvement in steatosis of liver tissue,a significant reduction in the percentage of oil red O staining area,significant reductions in the mRNA expression levels of Drp1 and Beclin-1,and a significant increase in the mRNA expression level of p62/SQSTM1(all P<0.05);there were significant reductions in the protein expression levels of Drp1,Beclin-1,and LC3BⅡ/Ⅰ,while some administration groups had a significant increase in the protein expression level of p62/SQSTM1(all P<0.05),with a significantly better effect in the middle-and high-dose Qizhu prescription groups(all P<0.01).Conclusion Qizhu prescription improves lipid metabolism and inflammation in mice with NAFLD possibly by regulating hepatocyte mitophagy. |
| Author | 陈佳豪 周振华 |
| AuthorAffiliation | 上海中医药大学附属曙光医院 细胞免疫实验室,上海 201203%上海中医药大学附属曙光医院肝病科,上海 201203;上海中医药大学附属曙光医院安徽医院肝病科,合肥 230011 |
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| Author_FL | CHEN Jiahao ZHOU Zhenhua |
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| Author_xml | – sequence: 1 fullname: 陈佳豪 – sequence: 2 fullname: 周振华 |
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| DocumentTitle_FL | Effect of Qizhu prescription on a mouse model of non-alcoholic fatty liver disease incluced by high-fat,high-fructose,and high-cholesterol diet and its mechanism |
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| Keywords | 非酒精性脂肪性肝病 芪术方 脂质代谢 Qi Zhu Formula 炎症 Non-alcoholic Fatty Liver Disease Lipid Metabolism Inflammation Mitophagy 线粒体自噬 |
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| PublicationTitle_FL | Journal of Clinical Hepatology |
| PublicationYear | 2024 |
| Publisher | Journal of Clinical Hepatology 上海中医药大学附属曙光医院 细胞免疫实验室,上海 201203%上海中医药大学附属曙光医院肝病科,上海 201203 上海中医药大学附属曙光医院安徽医院肝病科,合肥 230011 |
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| Snippet | 目的 探讨芪术方对非酒精性脂肪性肝病 (NAFLD) 小鼠模型的疗效及作用机制。 方法 60只雄性C57BL/6J小鼠随机分为正常组，模型组，芪术方低 (4.75 g/kg) 、中 (9.50 g/kg) 、高剂量组 (19.00 g/kg) ，多唏磷脂酰胆碱组 (磷脂组) (228 mg/kg)... 目的 探讨芪术方对非酒精性脂肪性肝病(NAFLD)小鼠模型的疗效及作用机制.方法 60只雄性C57BL/6J小鼠随机分为正常组,模型组,芪术方低(4.75 g/kg)、中(9.50 g/kg)、高剂量组(19.00 g/kg),多唏磷脂酰胆碱组(磷脂组)(228... |
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| SubjectTerms | Cholesterol Fatty liver Liver diseases Protein expression Proteins Tumor necrosis factor-TNF |
| Title | 芪术方对高脂高果糖高胆固醇诱导的非酒精性脂肪性肝病小鼠模型的影响及其机制 |
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