title: Hawthorn (Crataegus pinnatifida Bunge) leave flavonoids attenuate atherosclerosis development in apoE knock-out mice.
abstract: BACKGROUND
  Hawthorn (Crataegus pinnatifida Bunge) leave have been used to treat cardiovascular diseases in China and Europe. Hawthorn leave flavonoids (HLF) are the main part of extraction. Whether hawthorn leave flavonoids could attenuate the development of atherosclerosis and the possible mechanism remain unknown.
  
  
  METHODS
  High-fat diet (HFD) mixed with HLF at concentrations of 5mg/kg and 20mg/kg were administered to apolipoprotein E (apoE) knock out mice. 16 weeks later, mouse serum was collected to determine the lipid profile while the mouse aorta dissected was prepared to measure the lesion area. Hepatic mRNA of genes involved in lipid metabolism were determined. Peritoneal macrophages were collected to study the impact of HLF on cholesterol efflux, formation of foam cell and the expression of ATP binding cassette transporter A1 (ABCA1). Besides, in vivo reverse cholesterol transport (RCT) was conducted.
  
  
  RESULTS
  HLF attenuated the development of atherosclerosis that the mean atherosclerotic lesion area in en face aortas was reduced by 23.1% (P<0.05). In mice fed with 20mg/kg HLF, Total cholesterol (TC) level was decreased by 18.6% and very low density lipoprotein cholesterol plus low density lipoprotein cholesterol (VLDLc+LDLc) level were decreased by 23.1% whereas high density lipoprotein cholesterol (HDLc) and triglyceride (TG) levels were similar compared to that of the control group. Peroxisome proliferator activated receptor alpha (PPARα) mRNA was increased by 31.2% (P<0.05) and 60.9% (P<0.05) in mice fed with 5mg/kg and 20mg/kg HLF respectively. Sterol regulatory element binding protein-1c (SREBP-1c) was decreased by 59.3% in the group of 20mg/kg. Carnitine palmitoyl transferase 1 (CPT-1) mRNA level of 20mg/kg group was induced 66.7% (P<0.05). Superoxide dismutase 1 and 2 (SOD1 and SOD2) mRNA were induced 25.4% (P<0.05) and 71.4% (P<0.05) while induced by 36.3% (P<0.05) and 73.2% (P<0.05) in group of 20mg/kg. Glutathione peroxidase 3 (Gpx3) mRNA in the group of 20mg/kg was induced by 96.7% (P<0.05). Hepatic hydroxymethylglutaryl CoA reductase (HMG-CoAR) expression was as same level as the control group while LDL receptor (LDLR) mRNA and protein were induced by 84.2% (P<0.05) and 98.8% (P<0.05) in group of 20mg/kg. HLF inhibit the formation of foam cell by 27.9% (P<0.05) in the dosage of 25μg/ml, and 33.3% (P<0.05) in the dosage of 50μg/ml. HLF increased the reverse cholesterol transport (RCT) in vivo.
  
  
  CONCLUSIONS
  Hawthorn leave flavonoids can slow down the development of atherosclerosis in apoE knockout mice via induced expression of genes involved in antioxidant activities, inhibition of the foam cell formation and promotion of RCT in vivo, which implies the potential use in the prevention of atherosclerosis.
authors.affiliation: Tianjin State Key Laboratory of Modern Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin 300193, China; Research and Development Center of TCM, Tianjin International Joint Academy of Biotechnology & Medicine, Tianjin, China. Electronic address: pengzhiryn@163.com.
authors.forename: Pengzhi
authors.initials: P
authors.lastname: Dong
authors.affiliation: Tianjin State Key Laboratory of Modern Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin 300193, China; Research and Development Center of TCM, Tianjin International Joint Academy of Biotechnology & Medicine, Tianjin, China. Electronic address: 1587913321@qq.com.
authors.forename: Lanlan
authors.initials: L
authors.lastname: Pan
authors.affiliation: Tianjin State Key Laboratory of Modern Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin 300193, China; Research and Development Center of TCM, Tianjin International Joint Academy of Biotechnology & Medicine, Tianjin, China. Electronic address: 1655185436@qq.com.
authors.forename: Xiting
authors.initials: X
authors.lastname: Zhang
authors.affiliation: The College of Life Sciences, Nankai University, Tianjin 300071, China. Electronic address: 707991647@qq.com.
authors.forename: Wenwen
authors.initials: W
authors.lastname: Zhang
authors.affiliation: The Academy of Fundamental and Interdisciplinary Science, Harbin Institute of Technology, Harbin 150080, China. Electronic address: 664904176@qq.com.
authors.forename: Xue
authors.initials: X
authors.lastname: Wang
authors.affiliation: Institute of Translational Medicine, Nanchang University, Nanchang 330031, China. Electronic address: 191483003@qq.com.
authors.forename: Meixiu
authors.initials: M
authors.lastname: Jiang
authors.affiliation: School of Medicine, Nankai University, Tianjin 300071, China. Electronic address: chyuanli@163.com.
authors.forename: Yuanli
authors.initials: Y
authors.lastname: Chen
authors.affiliation: The College of Life Sciences, Nankai University, Tianjin 300071, China. Electronic address: yajunduan@nankai.edu.cn.
authors.forename: Yajun
authors.initials: Y
authors.lastname: Duan
authors.affiliation: Tianjin State Key Laboratory of Modern Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin 300193, China; Research and Development Center of TCM, Tianjin International Joint Academy of Biotechnology & Medicine, Tianjin, China. Electronic address: wuhonghua2003@163.com.
authors.forename: Honghua
authors.initials: H
authors.lastname: Wu
authors.affiliation: Tianjin State Key Laboratory of Modern Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin 300193, China; Research and Development Center of TCM, Tianjin International Joint Academy of Biotechnology & Medicine, Tianjin, China. Electronic address: xuyantong@gmail.com.
authors.forename: Yantong
authors.initials: Y
authors.lastname: Xu
authors.affiliation: Tianjin State Key Laboratory of Modern Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin 300193, China; Research and Development Center of TCM, Tianjin International Joint Academy of Biotechnology & Medicine, Tianjin, China. Electronic address: zhp8270@sina.com.cn.
authors.forename: Peng
authors.initials: P
authors.lastname: Zhang
authors.affiliation: Tianjin State Key Laboratory of Modern Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin 300193, China; Research and Development Center of TCM, Tianjin International Joint Academy of Biotechnology & Medicine, Tianjin, China. Electronic address: yanzhu.harvard@icloud.com.
authors.forename: Yan
authors.initials: Y
authors.lastname: Zhu
mesh_terms: D000818:Animals; D001011:Aorta / Q000187:drug effects / Q000473:pathology; D001057:Apolipoproteins E / Q000235:genetics*; D050197:Atherosclerosis / Q000235:genetics / Q000473:pathology / Q000517:prevention & control*; D002784:Cholesterol / Q000378:metabolism; D027825:Crataegus / Q000737:chemistry*; D059305:Diet, High-Fat; D005419:Flavonoids / Q000302:isolation & purification / Q000494:pharmacology*; D005487:Foam Cells / Q000166:cytology / Q000187:drug effects; D005786:Gene Expression Regulation / Q000187:drug effects; D050356:Lipid Metabolism / Q000187:drug effects / Q000235:genetics; D017737:Macrophages, Peritoneal / Q000187:drug effects / Q000378:metabolism; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D012333:RNA, Messenger / Q000378:metabolism
chemical_list: D001057:Apolipoproteins E; D005419:Flavonoids; D012333:RNA, Messenger; D002784:Cholesterol
keywords: Atherosclerosis; Foam cell; Hawthorn (Crataegus pinnatifida Bunge) leave flavonoids; Reverse cholesterol transport