1. Dingqiao Hospital of Hangzhou,Hangzhou 310021, China; 2. The First Affiliated Hospital of Zhejiang Chinese Medical University, Hangzhou 310006, China; 3. Key Laboratory of Circulatory System Disease Diagnosis and Treatment of Integrated Traditional Chinese and Western Medicine of Zhejiang, Hangzhou 310018, China
Abstract:OBJECTIVE To investigate the effect of pure total flavonoids from Citrus(PTFC) on the NLRP3 inflammasome pathway in mice with high-fat/high-fructose diet induced non-alcoholic steatohepatitis(NASH), and discuss its anti-NASH mechanism. METHODS Fifty C57BL/6 mice were randomly divided into normal group, model group, PTFC low-dosage group, medium-dosage group and high-dosage group, 10 in each group. A high-fat/high-fructose diet was given for 16 weeks to establish the NASH models, and the PTFC groups were administrated with PTFC at the dosage of 25, 50, 100 mg·kg-1·d-1 from the 5th week, respectively. Histopathologic changes of the liver tissue were observed by HE and oil red O staining; serum CHOL, TG, ALT, AST were detected by biochemical method; the levels of NLRP3, ASC, Casp1 and IL-1β mRNA in the liver tissue were determined by Realtime-PCR; the protein expression of NLRP3 and Casp1 was detected by Western blot. RESULTS Sixteen weeks administration of high-fat/high-fructose diet induced significantly higher weight, increased NAFLD activity score (NAS) of the liver tissue, higher level of serum CHOL, ALT and AST, lower level of serum TG, increased expression of NLRP3, ASC, Casp1 and IL-1β mRNA in liver, and increased protein expression of Casp1. After PTFC administration, the mice, especially the PTFC high dosage group, exhibited (compared with the model group) marked lower NAS, decreased serum ALT and AST level, increased TG level, impaired NLRP3, ASC, Casp1 and IL-1β mRNA expression, and decreased protein expression of NLRP3 and Casp1. CONCLUSION The NLRP3 inflammasome pathway may play a key role in the development and progress of NASH in mice, and PTFC prevents the development of NASH through suppressing this pathway.
俞建顺, 陈芝芸, 吴黎艳, 何蓓晖, 严茂祥, 蒋剑平. 胡柚皮黄酮对非酒精性脂肪性肝炎小鼠肝脏NLRP3炎症小体的影响[J]. 中国药学杂志, 2019, 54(24): 2076-2081.
YU Jian-shun, CHEN Zhi-yun, WU Li-yan,HE Bei-hui, YAN Mao-xiang, JIANG Jian-ping. Effect of Pure Total Flavonoids from Citrus on NLRP3 Inflammasome in Mice with Non-alcoholic Steatohepatitis. Chinese Pharmaceutical Journal, 2019, 54(24): 2076-2081.
MANNE V, HANDA P, KOWDLEY K V. Pathophysiology of nonalcoholic fatty liver disease/nonalcoholic steatohepatitis[J]. Clin Liver Dis, 2018,22(1):23-37.
[2]
MITSUYOSHI H, YASUI K, HARA T, et al. Hepatic nucleotide binding oligomerization domain-like receptors pyrin domain-containing 3 inflammasomes are associated with the histologic severity of non-alcoholic fatty liver disease[J]. Hepatol Res, 2017,47(13):1459-1468.
[3]
CHEN Z Y, LI J S, JIANG J P, et al. Effect of pure total flavonoids from Citrus on hepatic SIRT1/PGC-1α pathway in mice with NASH [J]. China J Chin Mater Med(中国中药杂志), 2014,39(1):100-105.
[4]
WU L, YAN M, JIANG J, et al. Pure total flavonoids from Citrus improve non-alcoholic fatty liver disease by regulating TLR/CCL signaling pathway: a preliminary high-throughput ′omics′ study[J]. Biomed Pharmacother, 2017, 93:316-326.
[5]
The Chinese National Workshop on Fatty Liver and Alcoholic Liver disease for the Chinese Liver Disease Association. Guidelines for management of nonalcoholic fatty liver disease:an updated and revised edition [J]. Chin J Hepatol(中华肝脏病杂志), 2010, 18(3):163-166.
[6]
DAY C P, JAMES O F. Steatohepatitis: a tale of two “hits”?[J]. Gastroenterology, 1998, 114(4):842-845.
[7]
SZABO G, CSAK T. Inflammasomes in liver diseases[J]. J Hepatol, 2012, 57(3): 642-654.
[8]
CSAK T, GANZ M, PESPISA J, et al. Fatty acid and endotoxin activate inflammasomes in mouse hepatocytes that release danger signals to stimulate immune cells[J]. Hepatology, 2011, 54(1):133-144.
[9]
DIXON L J, BERK M, THAPALIYA S, et al. Caspase-1-mediated regulation of fibrogenesis in diet-induced steatohepatitis[J]. Lab Invest, 2012, 92(5):713-723.
[10]
WU X, DONG L, LIN X, et al. Relevance of the NLRP3 inflammasome in the pathogenesis of chronic liver disease[J]. Front Immunol, 2017, 8:1728.
[11]
WREE A, EGUCHI A, MCGEOUGH M D, et al. NLRP3 inflammasome activation results in hepatocyte pyroptosis, liver inflammation, and fibrosis in mice[J]. Hepatology, 2014, 59(3): 898-910.
[12]
ALEGRE F, PELEGRIN P, FELDSTEIN A E. Inflammasomes in liver fibrosis[J]. Semin Liver Dis, 2017, 37(2):119-127.
[13]
SCHRODER K, TSCHOPP J. The inflammasomes[J]. Cell, 2010, 140(6): 821-832.
[14]
SNODGRASS R G, HUANG S, CHOI I W, et al. Inflammasome-mediated secretion of IL-1beta in human monocytes through TLR2 activation; modulation by dietary fatty acids[J]. J Immunol, 2013, 191(8):4337-4347.
[15]
YI Y S. Roles of ginsenosides in inflammasome activation[J]. J Ginseng Res, 2019, 43(2):172-178.
[16]
ZHAO X J, YANG Y Z, ZHENG Y J, et al. Magnesium isoglycyrrhizinate blocks fructose-induced hepatic NF-kappaB/NLRP3 inflammasome activation and lipid metabolism disorder[J]. Eur J Pharmacol, 2017, 809:141-150.
[17]
COBBINA E, AKHLAGHI F. Non-alcoholic fatty liver disease (NAFLD) - pathogenesis, classification, and effect on drug metabolizing enzymes and transporters[J]. Drug Metab Rev, 2017, 49(2):197-211.
[18]
JIANG J, SHAN L, CHEN Z, et al. Evaluation of antioxidant-associated efficacy of flavonoid extracts from a traditional Chinese medicine Hua Ju Hong [peels of Citrus grandis (L.) Osbeck][J]. J Ethnopharmacol, 2014, 158 Pt A: 325-330.
[19]
LI J S, CHEN Z Y, JIANG J P, et al. Regulation of pure total flavonoids from Citrus on TH17/Treg balance in mice with NASH [J]. China J Chin Mater Med(中国中药杂志), 2015,40(13):2644-2648.