title: Protective Effect of Ganshuang Granules () on Liver Cirrhosis by Suppressing Regulatory T Cells in Mouse Model.
abstract: OBJECTIVE
  To investigate the potential antifibrotic mechanisms of Chinese medicine Ganshuang Granules (, GSG) and to provide clinical therapeutic evidence of its effects.
  
  
  METHODS
  A cirrhotic mouse model was established by intraperitoneally injecting a mixture of CCl4(40%) and oil (60%) at 0.2 mL per 100 g of body weight twice a week for 12 weeks. After 12-week modeling, GSG was intragastric administrated to the mice for 2 weeks, and the mice were divided into low-, medium- and high-dose groups at doses of 1, 2 and 4 g/(kg·day), respectively. Liver morphology changes were observed using Masson's trichrome staining and B-ultrasound. The levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and hyaluronic acid (HA) in serum were detected using an automatic biochemistry analyzer. The expressions of desmin, smooth muscle actin (SMA) and Foxp3 in liver were detected by immunoflfluorescence. The regulatory T cell (Treg) frequency was determined through flflow cytometry analysis. Collagen-I, SMA, IL-6, tumor necrosis factor α (TNF-α), interleukin (IL)-1β and transforming growth factor β1 (TGF-β1) expression levels were measured using quantitative polymerase chain reaction (qPCR).
  
  
  RESULTS
  Masson's staining result showed fewer pseudolobule structures and fibrous connective tissue in the GSG-treatment groups than in the spontaneous recovery group. Ultrasonography showed that GSG treatment reduced the number of punctate hyperechoic lesions in mice cirrhotic livers. The serum ALT, AST, HA levels were significantly ameliorated by GSG treatment (ALT: F=8.104, P=0.000; AST: F=7.078, P=0.002; and HA: F=7.621, P=0.001). The expression levels of collagen-I and SMA in the cirrhotic livers were also attenuated by GSG treatment (collagen-I: F=3.938, P=0.011; SMA: F=4.115, P=0.009). Tregs, which were elevated in the fibrotic livers, were suppressed by GSG treatment (F=8.268, P=0.001). The expressions of IL-6, TNF-α and IL-1β increased, and TGF-β levels decreased in the cirrhotic livers after GSG treatment (IL-6: F=5.457, P=0.004; TNF-α: F=6.023, P=0.002; IL-1β: F=6.658, P=0.001; and TGF-β1: F=11.239, P=0.000).
  
  
  CONCLUSIONS
  GSG promoted the resolution/regression of cirrhosis and restored liver functions in part by suppressing Treg cell differentiation, which may be mediated by hepatic stellate cells.
authors.affiliation: Department of Liver Diseases Immunology, Beijing Youan Hospital, Capital Medical University, Beijing, 100069, China.
authors.forename: Yan-Min
authors.initials: YM
authors.lastname: Liu
authors.affiliation: Department of Liver Diseases Immunology, Beijing Youan Hospital, Capital Medical University, Beijing, 100069, China.|Beijing Institute of Hepatology, Beijing Youan Hospital, Capital Medical University, Beijing, 100069, China.
authors.forename: Hong-Bo
authors.initials: HB
authors.lastname: Shi
authors.affiliation: Department of Toxic Hepatic Disease, Beijing Youan Hospital, Capital Medical University, Beijing, 100069, China.
authors.forename: Yi-Rong
authors.initials: YR
authors.lastname: Liu
authors.affiliation: Department of Liver Diseases Immunology, Beijing Youan Hospital, Capital Medical University, Beijing, 100069, China.|Beijing Institute of Hepatology, Beijing Youan Hospital, Capital Medical University, Beijing, 100069, China.
authors.forename: Hong-Lin
authors.initials: HL
authors.lastname: Shi
authors.affiliation: Department of Liver Diseases Immunology, Beijing Youan Hospital, Capital Medical University, Beijing, 100069, China.|Beijing Institute of Hepatology, Beijing Youan Hospital, Capital Medical University, Beijing, 100069, China.
authors.forename: Feng
authors.initials: F
authors.lastname: Ren
authors.affiliation: Artifical Liver Center, Beijing Youan Hospital, Capital Medical University, Beijing, 100069, China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Chen
authors.affiliation: Department of Liver Diseases Immunology, Beijing Youan Hospital, Capital Medical University, Beijing, 100069, China.|Beijing Institute of Hepatology, Beijing Youan Hospital, Capital Medical University, Beijing, 100069, China.
authors.forename: De-Xi
authors.initials: DX
authors.lastname: Chen
authors.affiliation: Clinical Laboratory Center, Beijing Youan Hospital, Capital Medical University, Beijing, 100069, China.
authors.forename: Jin-Li
authors.initials: JL
authors.lastname: Lou
authors.affiliation: Artifical Liver Center, Beijing Youan Hospital, Capital Medical University, Beijing, 100069, China. duan2517@163.com.
authors.forename: Zhong-Ping
authors.initials: ZP
authors.lastname: Duan
mesh_terms: D000818:Animals; D004195:Disease Models, Animal; D004365:Drugs, Chinese Herbal / Q000494:pharmacology* / Q000627:therapeutic use; D055166:Hepatic Stellate Cells / Q000187:drug effects; D008106:Liver Cirrhosis, Experimental / Q000188:drug therapy* / Q000276:immunology; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D050378:T-Lymphocytes, Regulatory / Q000187:drug effects*
chemical_list: D004365:Drugs, Chinese Herbal
keywords: Chinese medicine; Ganshuang Granules; hepatic stellate cell; liver cirrhosis; regulatory T cell