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Anandamide induces cell death through lipid rafts in hepatic stellate cells.
J Gastroenterol Hepatol. 2010 May; 25(5):991-1001.JG

Abstract

BACKGROUND AND AIMS

Anandamide (AEA), the most extensively studied endocannabinoid, and its putative cannabinoid receptors, CB1 and CB2, exert a variety of physiological and pharmacological effects in chronic liver diseases, such as hyperdynamic circulation. Anandamide selectively blocks proliferation and induces cell death in hepatic stellate cells (HSC), the key cell type of liver fibrogenesis. However, its precise molecular mechanism in rat HSC has not been fully elucidated.

METHODS

CB1 and CB2 mRNA transcriptions were evaluated by reverse transcription polymerase chain reaction; CB1, CB2, phosphoinositide 3-kinases (PI3K) and protein kinase B (PKB) protein expressions were investigated by western blot and/or immunofluorescence. Cell death was detected by Annexin V-PE/7AAD flow cytometry, lipid raft content by confocal microscopic analysis, cell viability by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay, nuclear morphological changes by Hoechst 33258 fluorochrome, and inflammatory cytokines interleukin (IL)-2 and IL-6, and tumor necrosis factor-alpha (TNF-alpha) by enzyme-linked immunosorbent assay.

RESULTS

CB1 and CB2 receptors were detectable in HSC. AEA caused HSC growth inhibition in a concentration-dependent manner. Furthermore, a high concentration of AEA (20 micromol/L) triggered potent cell death-induced necrosis but not apoptosis. None of these effects were blocked by CB1 or CB2 receptor antagonist, but by methyl-beta-cyclodextrin (MCD; 10 mmol/L), a cholesterol depletory agent. AEA significantly inhibited PI3K/PKB activity, and increased IL-2, IL-6 and TNF-alpha release.

CONCLUSION

These results demonstrated that AEA induced HSC necrosis through lipid rafts: a possible role of PI3K/PKB signaling pathway downregulation and inflammatory factors production. Cholesterol depletion abolished the effects of AEA on HSC necrosis.

Authors+Show Affiliations

Institute of Liver Diseases, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

20059638

Citation

Yang, Qiao, et al. "Anandamide Induces Cell Death Through Lipid Rafts in Hepatic Stellate Cells." Journal of Gastroenterology and Hepatology, vol. 25, no. 5, 2010, pp. 991-1001.
Yang Q, Liu HY, Zhang YW, et al. Anandamide induces cell death through lipid rafts in hepatic stellate cells. J Gastroenterol Hepatol. 2010;25(5):991-1001.
Yang, Q., Liu, H. Y., Zhang, Y. W., Wu, W. J., & Tang, W. X. (2010). Anandamide induces cell death through lipid rafts in hepatic stellate cells. Journal of Gastroenterology and Hepatology, 25(5), 991-1001. https://doi.org/10.1111/j.1440-1746.2009.06122.x
Yang Q, et al. Anandamide Induces Cell Death Through Lipid Rafts in Hepatic Stellate Cells. J Gastroenterol Hepatol. 2010;25(5):991-1001. PubMed PMID: 20059638.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Anandamide induces cell death through lipid rafts in hepatic stellate cells. AU - Yang,Qiao, AU - Liu,Hong Yan, AU - Zhang,Yao Wen, AU - Wu,Wen Jie, AU - Tang,Wang Xian, Y1 - 2010/01/04/ PY - 2010/1/12/entrez PY - 2010/1/12/pubmed PY - 2010/9/29/medline SP - 991 EP - 1001 JF - Journal of gastroenterology and hepatology JO - J Gastroenterol Hepatol VL - 25 IS - 5 N2 - BACKGROUND AND AIMS: Anandamide (AEA), the most extensively studied endocannabinoid, and its putative cannabinoid receptors, CB1 and CB2, exert a variety of physiological and pharmacological effects in chronic liver diseases, such as hyperdynamic circulation. Anandamide selectively blocks proliferation and induces cell death in hepatic stellate cells (HSC), the key cell type of liver fibrogenesis. However, its precise molecular mechanism in rat HSC has not been fully elucidated. METHODS: CB1 and CB2 mRNA transcriptions were evaluated by reverse transcription polymerase chain reaction; CB1, CB2, phosphoinositide 3-kinases (PI3K) and protein kinase B (PKB) protein expressions were investigated by western blot and/or immunofluorescence. Cell death was detected by Annexin V-PE/7AAD flow cytometry, lipid raft content by confocal microscopic analysis, cell viability by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay, nuclear morphological changes by Hoechst 33258 fluorochrome, and inflammatory cytokines interleukin (IL)-2 and IL-6, and tumor necrosis factor-alpha (TNF-alpha) by enzyme-linked immunosorbent assay. RESULTS: CB1 and CB2 receptors were detectable in HSC. AEA caused HSC growth inhibition in a concentration-dependent manner. Furthermore, a high concentration of AEA (20 micromol/L) triggered potent cell death-induced necrosis but not apoptosis. None of these effects were blocked by CB1 or CB2 receptor antagonist, but by methyl-beta-cyclodextrin (MCD; 10 mmol/L), a cholesterol depletory agent. AEA significantly inhibited PI3K/PKB activity, and increased IL-2, IL-6 and TNF-alpha release. CONCLUSION: These results demonstrated that AEA induced HSC necrosis through lipid rafts: a possible role of PI3K/PKB signaling pathway downregulation and inflammatory factors production. Cholesterol depletion abolished the effects of AEA on HSC necrosis. SN - 1440-1746 UR - https://www.unboundmedicine.com/medline/citation/20059638/Anandamide_induces_cell_death_through_lipid_rafts_in_hepatic_stellate_cells_ L2 - https://doi.org/10.1111/j.1440-1746.2009.06122.x DB - PRIME DP - Unbound Medicine ER -