Inhibitory effects of Lobelia Chinensis Lour alkaloid on the proliferation of human umbilical artery vascular smooth muscle induced by endothelin-1
HU Wei-cheng
Abstract
HU Wei-cheng
Abstract
AIM: To investigate the effects of Lobelia Chinensis Lour alkaloid (LCLA) on the proliferation of cultured vascular smooth muscle cells (VSMC) induced by ET-1. METHODS: Human umbilical artery VSMC was cultured and divided into five groups: ET group, ET+LCLA group, ET+BQ-123 group,ET+ staurosporine (ST) group and control group. The cell proliferation activity was subsequently quantified by cell counting kit-8 (CCK-8) and [3H]-TdR incorporation. Flow cytometry was used to examine cell cycle. Quantitative immunohistochemical technique was used to investigate the expression of proliferating cell nuclear antigen (PCNA) and confocal microscope was used to measure the fluorescent intensity of Ca 2+. Cytotoxicity was measured by Trypan blue exclusion and LDH colorimetry tests. RESULTS: BQ-123 (10 -6mol/L), ST (10 -7mol/L) and LCLA (100, 200 and 400 mg/L) inhibited the increase in cell number, [3H]-TdR incorporation, the percentage of the S phase and markedly decreased the expression of PCNA and fluorescent intensity of Ca 2+ in response to ET-1 of VSMC (P0.05). CONCLUSION: LCLA (100-400 mg/L) inhibits ET-1-induced proliferation of VSMC in a dose-dependent manner and the anti-proliferative effect is realized by reducing the Ca 2+ concentration in VSMC.
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AIM: To investigate the effects of Lobelia Chinensis Lour alkaloid (LCLA) on the proliferation of cultured vascular smooth muscle cells (VSMC) induced by ET-1. METHODS: Human umbilical artery VSMC was cultured and divided into five groups: ET group, ET+LCLA group, ET+BQ-123 group,ET+ staurosporine (ST) group and control group. The cell proliferation activity was subsequently quantified by cell counting kit-8 (CCK-8) and [3H]-TdR incorporation. Flow cytometry was used to examine cell cycle. Quantitative immunohistochemical technique was used to investigate the expression of proliferating cell nuclear antigen (PCNA) and confocal microscope was used to measure the fluorescent intensity of Ca 2+. Cytotoxicity was measured by Trypan blue exclusion and LDH colorimetry tests. RESULTS: BQ-123 (10 -6mol/L), ST (10 -7mol/L) and LCLA (100, 200 and 400 mg/L) inhibited the increase in cell number, [3H]-TdR incorporation, the percentage of the S phase and markedly decreased the expression of PCNA and fluorescent intensity of Ca 2+ in response to ET-1 of VSMC (P0.05). CONCLUSION: LCLA (100-400 mg/L) inhibits ET-1-induced proliferation of VSMC in a dose-dependent manner and the anti-proliferative effect is realized by reducing the Ca 2+ concentration in VSMC.
Key concepts: Vascular smooth muscle, Proliferating cell nuclear antigen, Flow cytometry, Molecular biology, Cell growth, Cell cycle, Alkaloid, Trypan blue