ABSTRACT
AIM: To investigate the effects of triptolide on vascular endothelial growth factor (VEGF) expression and secretion by endothelial cells, and explore the mechanism of anti-proteinuric effect of triptolide on glomerulonephritis. METHODS: A human umbilical endothelium derived cell line (ECV-304) from American Type Culture Collection (ATCC) was used in this study. The effects of triptolide on VEGF mRNA expression, production, and secretion induced by 12-o-tetradecanoyl-phorbol-13-acetate (TPA) were measured by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR), flow cytometry, and enzyme linked immunosorbent assay (ELISA) respectively. The endothelial c-fos/c-jun mRNA expression were also detected by RT-PCR after treatment of triptolide. RESULTS: VEGF mRNA expression was markedly up-regulated by TPA-stimulation. In addition, the production and secretion of VEGF in endothelial cells also increased in TPA treated cells. It was founded that triptolide inhibited VEGF mRNA expression, protein production and secretion in endothelial cells induced by TPA. Interestingly, TPA-induced c-fos/c-jun mRNA expression in endothelial cells was also inhibited by triptolide. CONCLUSION: Triptolide is a potent inhibitor of VEGF expression and production in endothelial cells. The inhibitory effects of triptolide on VEGF expression and production can contribute to its anti-proteinuric effect on glomerulonephritis. Down-regulation of c-fos/c-jun expression in endothelial cells by triptolide is one of the mechanisms of the inhibitory effect of triptolide on VEGF expression.
Subject(s)
Diterpenes/pharmacology , Endothelial Growth Factors/biosynthesis , Endothelium, Vascular/metabolism , Intercellular Signaling Peptides and Proteins/biosynthesis , Lymphokines/biosynthesis , Phenanthrenes , Anti-Inflammatory Agents, Non-Steroidal/pharmacology , Cells, Cultured , Endothelial Growth Factors/genetics , Endothelium, Vascular/cytology , Epoxy Compounds , Humans , Intercellular Signaling Peptides and Proteins/genetics , Lymphokines/genetics , Proto-Oncogene Proteins c-fos/biosynthesis , Proto-Oncogene Proteins c-fos/genetics , Proto-Oncogene Proteins c-jun/biosynthesis , Proto-Oncogene Proteins c-jun/genetics , RNA, Messenger/biosynthesis , RNA, Messenger/genetics , Umbilical Veins/cytology , Vascular Endothelial Growth Factor A , Vascular Endothelial Growth FactorsABSTRACT
Alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH), the principal enzymes responsible for oxidative metabolism of ethanol, exist in multiple, genetically determined molecular forms. Widely different kinetic properties in some of these isozymes account for the individual differences in alcohol sensitivity. In this study we used the polymerase chain reaction/restriction fragment length polymorphism method to determine the genotypes of the ADH2 and ALDH2 loci of alcoholic and nonalcoholic Chinese living in Shanghai. We also investigated the subjects' drinking patterns by means of semistructured interviews. The alcoholics had significantly lower frequencies of the ADH2(2) and ALDH2(2) alleles than did the nonalcoholics, suggesting the inhibitory effects of these alleles for the development of alcoholism. In the nonalcoholic subjects, ADH2(2) had little, if any, effect, despite the significant effect of the ALDH2(2) allele in decreasing the alcohol consumption of the individual. Taken together, these results fit the proposed hypothesis for the development of alcoholism, i.e., drinking behavior is greatly influenced by the individual's genotypes of alcohol-metabolizing enzymes, and the risk of becoming alcoholic is proportionate with the ethanol consumption of the individual.
