title: Xiayuxue Decoction ([symbols; see text]) attenuates hepatic stellate cell activation and sinusoidal endothelium defenestration in CCl4-induced fibrotic liver of mice.
abstract: OBJECTIVE
  To investigate the effects of ancient Chinese medical formula Xiayuxue Decoction ([symbols; see text], XYXD) on activation of hepatic stellate cells (HSCs) and defenestration of sinusoidal endothelial cells (SECs) in CCl4-induced fibrotic liver of mice.
  
  
  METHODS
  High performance liquid chromatography was used to identify the main components of XYXD and control the quality of extraction. C57BL/6 mice were induced liver fibrosis by CCl4 exposure and administered with XYXD for 6 weeks simultaneously. Liver tissue was investigated by hematoxylin-eosin and Sirius-red staining. Sinusoidal fenestrations were observed by scanning electronic microscopy and fluorescent immunohistochemistry of PECAM-1 (CD31). Whole liver lysates were detected of α-smooth muscle actin (α-SMA) and type-I collagen by Western blot. Primary rat HSCs-T6 cells were analyzed by detecting α-SMA, F-actin, DNA fragmentation through confocal microscopy, Western blot, terminal-deoxynucleoitidyl transferase mediated nick end labeling (TUNEL) assay and cellomics arrayscan, respectively.
  
  
  RESULTS
  Amygdalin and emodin in XYXD were identified. XYXD (993 mg/kg) inhibited Sirius red positive area up to 70.1% (P<0.01), as well as protein levels of α-SMA and type-I collagen by 42.0% and 18.5% (P<0.05) respectively. In vitro, XYXD (12.5 μg/mL, 50 μg/mL) suppressed the activation of HSCs and reversed the myofibroblastic HSCs into quiescent, demonstrated as inhibition of fluorescent F-actin by 32.3% and 46.6% (P<0.05). Besides, XYXD induced the apoptosis of HSC-T6 cells by 20.0% (P<0.05) and 49.5% (P<0.01), evidenced by enhanced TUNEL positivity. Moreover, ultrastructural observation suggested XYXD inhibited defenestration of SECs, which was confirmed by 31.1% reduction of protein level of CD31 (P<0.05).
  
  
  CONCLUSIONS
  XYXD inhibited both HSCs activation and SECs defenestration which accompany chronic liver injuries. These data may help to understand the underlying mechanisms of XYXD for prevetion of chronic liver diseases.
authors: Institute of Liver Diseases, Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai, 201203, China.
authors: Li-jun
authors: LJ
authors: Zhang
authors: Ming-yu
authors: MY
authors: Sun
authors: Bing-bing
authors: BB
authors: Ning
authors: Wen-meng
authors: WM
authors: Zhang
authors: Gao-feng
authors: GF
authors: Chen
authors: Yong-ping
authors: YP
authors: Mu
authors: Hua
authors: H
authors: Zhang
authors: Jia
authors: J
authors: Liu
authors: Yan-qin
authors: YQ
authors: Bian
authors: Ping
authors: P
authors: Liu
mesh_terms: D000199:Actins / Q000378:metabolism; D000818:Animals; D002252:Carbon Tetrachloride Poisoning / Q000188:drug therapy; D024042:Collagen Type I / Q000378:metabolism; D004195:Disease Models, Animal; D004365:Drugs, Chinese Herbal / Q000494:pharmacology*; D004727:Endothelium / Q000187:drug effects / Q000473:pathology; D055166:Hepatic Stellate Cells / Q000187:drug effects* / Q000473:pathology / Q000648:ultrastructure; D008103:Liver Cirrhosis / Q000139:chemically induced / Q000188:drug therapy* / Q000473:pathology; D008297:Male; D008810:Mice, Inbred C57BL; D008855:Microscopy, Electron, Scanning; D058628:Myofibroblasts / Q000187:drug effects / Q000473:pathology / Q000648:ultrastructure; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000378:metabolism; D061251:Primary Cell Culture; D017207:Rats, Sprague-Dawley
chemical_list: D000199:Actins; D024042:Collagen Type I; D004365:Drugs, Chinese Herbal; D019408:Platelet Endothelial Cell Adhesion Molecule-1