Protective effect of JBP485 on concanavalin A-induced hepatocyte toxicity in primary cultured rat hepatocytes.
Cyclo-trans-4-L-hydroxyprolyl-L-serine (JBP485) is a dipeptide isolated from Laennec, and Laennec is a hydrolyzate of human placenta. Evidence has indicated that JBP485 exhibits potent anti-hepatitis activity. In this study, we investigated the protective effect and possible mechanisms of action of JBP485 in Concanavalin A (Con A)-induced hepatotoxicity in vitro. Two in vitro models were established. Model I: primary cultured female rat hepatocytes were only incubated with Con A (50 microg/ml); model II: co-culture system of hepatocytes and autologous splenic lymphocytes, both were stimulated with Con A (20 microg/ml). JBP485 (25 microM) was pre-incubated with the two models. Our results showed that JBP485 reduced cellular aspartate aminotransferase (AST), lactate dehydrogenase (LDH) and tumor necrosis factor alpha (TNF-alpha) leakage following the application of Con A in both of the models. Potential protective mechanisms were elucidated by measuring DNA fragmentations, immunocytochemistry and RT-PCR. We showed that DNA fragmentations in hepatocytes were attenuated in the JBP485 pre-incubated groups, and at the same time, immunocytochemistry and RT-PCR indicated that expression levels of caspase-3 protein and mRNA in the JBP485 treated groups were decreased compared with those in the untreated groups. Moreover, intercellular adhesion molecule-1 (ICAM-1) was also down-regulated by this dipeptide. The results indicate that JBP485 exhibits hepatoprotective effect through inhibition of hepatocyte apoptosis and ICAM-1 expression.
Department of Clinical Pharmacology, College of Pharmacy, Dalian Medical University, Liaoning, Dalian, China.
Jingjing
J
Wu
Changyuan
C
Wang
Qi
Q
Liu
Tao
T
Yang
Qinghao
Q
Zhang
Jinyong
J
Peng
Ying
Y
Gao
Huijun
H
Sun
Taiichi
T
Kaku
Kexin
K
Liu
D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D001219:Aspartate Aminotransferases / Q000378:metabolism; D053148:Caspase 3 / Q000235:genetics / Q000378:metabolism; D018929:Cell Culture Techniques; D002478:Cells, Cultured; D018920:Coculture Techniques; D003208:Concanavalin A / Q000633:toxicity*; D019610:Cytoprotection; D053938:DNA Fragmentation; D004305:Dose-Response Relationship, Drug; D005260:Female; D022781:Hepatocytes / Q000187:drug effects* / Q000201:enzymology / Q000276:immunology / Q000473:pathology; D007150:Immunohistochemistry; D018799:Intercellular Adhesion Molecule-1 / Q000378:metabolism; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008214:Lymphocytes / Q000187:drug effects; D010456:Peptides, Cyclic / Q000494:pharmacology*; D020011:Protective Agents / Q000494:pharmacology*; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
C412308:JBP 485; D010456:Peptides, Cyclic; D020011:Protective Agents; D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha; D003208:Concanavalin A; D018799:Intercellular Adhesion Molecule-1; D007770:L-Lactate Dehydrogenase; D001219:Aspartate Aminotransferases; C505410:Casp3 protein, rat; D053148:Caspase 3