title: Combined effects of an antioxidant and caspase inhibitor on the reversal of hepatic fibrosis in rats.
abstract: We sought to determine the hepatic fibrosis-reversal effects upon simultaneous administration of lithospermate B (LAB), an anti-oxidant, and nivocasan, a caspase inhibitor, to rats compared with each compound alone. Liver fibrosis was induced in Sprague-Dawley rats by thioacetamide (TAA). Rats were treated with TAA and then given LAB and (or) nivocasan. Fibrotic areas were evaluated quantitatively by computerized morphometry. Apoptosis was assessed using a TUNEL assay, and immunohistochemical staining for malondialdehyde (MDA) and 4-hydroxy-2-nonenal (4HNE) was performed to assess oxidative stress levels. Real-time quantitative PCR was used to quantify expression of fibrosis-related genes. The degree of hepatic fibrosis was significantly reduced in rats treated with LAB and nivocasan compared to either treatment alone (P < 0.001). Treatment with each compound significantly decreased expression of fibrosis-related genes, such as type I collagen α1 (col1α1), α-SMA and TGF-β1 (P < 0.05). Co-treatment with LAB and nivocasan further reduced col1α1 expression compared to treatment with either compound. A TUNEL assay revealed that hepatocyte apoptosis was significantly decreased in the group treated with nivocasan compared to other groups (P < 0.01). Immunohistochemistry showed a decrease in MDA and 4HNE, reflecting amelioration of oxidative stress, when LAB or LAB+nivocasan was administered compared to nivocasan alone (P < 0.01). Nivocasan was found to inhibit caspase-1, -3, -7, -9 and gliotoxin-induced death of rat-derived hepatic stellate cells was inhibited by nivocasan administration without overexpression of α-SMA.
  
  
  CONCLUSIONS
  Co-incidental administration of LAB and nivocasan suppressed oxidative stress and apoptosis, resulting in enhanced reversal of hepatic fibrosis in rat.
authors.affiliation: Department of Internal Medicine, Yonsei University College of Medicine, 50 Yonsei-ro, Seodaemun-gu, Seoul, 120-752, Korea, dyk1025@yuhs.ac.
authors.forename: Do Young
authors.initials: DY
authors.lastname: Kim
authors.forename: Sook In
authors.initials: SI
authors.lastname: Chung
authors.forename: Simon Weonsang
authors.initials: SW
authors.lastname: Ro
authors.forename: Yong Han
authors.initials: YH
authors.lastname: Paik
authors.forename: Jung Il
authors.initials: JI
authors.lastname: Lee
authors.forename: Man Kil
authors.initials: MK
authors.lastname: Jung
authors.forename: Min Goo
authors.initials: MG
authors.lastname: Lee
authors.forename: Young Nyun
authors.initials: YN
authors.lastname: Park
authors.forename: Kwan Sik
authors.initials: KS
authors.lastname: Lee
authors.forename: Jung Gyu
authors.initials: JG
authors.lastname: Park
authors.forename: Hee Dong
authors.initials: HD
authors.lastname: Park
authors.forename: Kwang-Hyub
authors.initials: KH
authors.lastname: Han
mesh_terms: D000818:Animals; D000975:Antioxidants / Q000008:administration & dosage*; D017209:Apoptosis / Q000187:drug effects*; D061945:Caspase Inhibitors / Q000008:administration & dosage*; D020169:Caspases / Q000235:genetics / Q000378:metabolism; D024042:Collagen Type I / Q000235:genetics / Q000378:metabolism; D000090602:Collagen Type I, alpha 1 Chain; D004359:Drug Therapy, Combination; D004365:Drugs, Chinese Herbal / Q000008:administration & dosage*; D006801:Humans; D008103:Liver Cirrhosis / Q000188:drug therapy* / Q000235:genetics / Q000378:metabolism / Q000503:physiopathology; D008297:Male; D018384:Oxidative Stress / Q000187:drug effects; D051381:Rats; D017207:Rats, Sprague-Dawley; D053773:Transforming Growth Factor beta1 / Q000235:genetics / Q000378:metabolism
chemical_list: D000975:Antioxidants; D061945:Caspase Inhibitors; D024042:Collagen Type I; D000090602:Collagen Type I, alpha 1 Chain; D004365:Drugs, Chinese Herbal; D053773:Transforming Growth Factor beta1; C061091:lithospermate B; D020169:Caspases