RESUMO
Objective To evaluate effects of Lycium barbarum polysaccharide (LBP) on expression of vascular endothelial growth factor (VEGF) and hypoxia inducible factor-1α (HIF-1α) in ultraviolet B (UVB)-radiated HaCaT cells.Methods Conventionally cultured HaCaT cells were divided into control group and LBP groups,which were firstly treated with DMEM,12.5,25.0,50.0 and 100 μg/ml LBP solution respectively for 4 hours,and then were irradiated by UVB at different intensity of 0,20,40,60 mJ/cm2 separately.After 24-hour continuing culture,CCK-8 assay was performed to determine the cell survival rate,and an enzymatic-biochemical method to estimate the activity of superoxide dismutase (SOD).RT-PCR and Western blot analysis were conducted to measure the mRNA and protein expression of HIF-1α and VEGF respectively.Results Compared with the control group at the same UVB radiation dose,the 12.5-,25.0-and 100.0-μ,g/ml LBP groups showed different extents of increase in survival rates of UVB-radiated cells (P ＜ 0.05),and the 50.0-μg/ml LBP group showed the highest cell survival rate (P ＜ 0.01).Among all the LBP groups,SOD activity was highest in the 50.0-μg/ml LBP group (P ＜ 0.01).Along with the increase of UVB radiation dose,the mRNA and protein expression of HIF-1α and VEGF all gradually increased.Compared with the control group,the 50.0-μg/ml LBP group could effectively reduce the mRNA and protein expression of HIF-1α and VEGF in HaCaT cells (all P ＜ 0.05).Conclusion LBP may play a role in protecting cells from UVB radiation-mediated damage,likely by influencing the mRNA and protein expression of HIF-1α and VEGF in HaCaT cells.
RESUMO
Objective: To investigate SIN ( Sinomenine) for TLR signal transduction pathway and MyD88 ( MyeloidDifferentiation Factor 88), TRAF-6 ( Tumor necrosis factor receptor associated factor-6) expression, clarifying SIN inhibit RA(Rheumatoid arthritis)-FLS(Fibroblast-like synoviocytes)proliferation leads to joint deformity of RA cartilage and subchondral bonedestruction caused by the role of mechanisms.Methods: RA-FLS cells for vitro were divided into a control group and(0.125,0.25,0.5,1 mmol/L)SIN group,within each group were detected by alkaline phosphatase(ALP)activity,to determine the best concentrationfor vitro drug;detect 0.5 mmol/L SIN group in CCK-8 method to detect cell proliferation rate;fluorescence quantitative PCR methodMyD88 SIN group and control group with 0.5 mmol/L TRAF-6 gene expression;Western blot method to detect MyD88 SIN group andcontrol group with 0.5 mmol/L TRAF-6 protein expression.Results: SIN the ALP activity of the lower than the control group,with theminimum ALP activity of 0.5 mmol/L SIN group(P<0.01).CCK 8 method,0.5 mmol/L RA-FLS cell proliferation rate SIN group wasobviously lower than the control group(P<0.01),SIN induce cell proliferation rate was highest,4 days into the plateau,after cell proliferationrate began to fall.Fluorescence quantitative PCR and Western blot method to detect,0.5 mmol/L SIN group of MyD88 andTRAF-6 gene and protein expression significantly lower than the control group,the difference was statistically significant(P<0.01). Conclusion: The SIN of TLR signalling pathways through effectively suppress the influence of RA-FLS MyD88 in the cell and theexpression of TRAF-6,this could be a treatment of RA prevent cartilage and subchondral bone damage cause joint deformity happened one of the important molecular mechanism.
RESUMO
Objective To study the influence of tripterygium glycosides (LGTDG)in the bone morphogenetic protein(BMP)signal transduction pathway and BMP-2 expression,and to clarify the mechanism of anti-ankylosing spondylitis (AS)ossification of LGTDG.Methods The in vitro cultured AS fibroblasts were divided into control group and 0.5,1.0,1.5,2.0 mg·L-1 LGTDG groups.The alkaline phosphatase (ALP)activities of the cells and optimal drug concentrations in various groups were detected;CCK-8 assay was used to detect the proliferation rate of the cells in 1.0 mg· L-1 LGTDG group;the biochemical tests were performed to quantitatively detect the BMP-2 expression levels in control group and 1.0 mg· L-1 LGTDG group;Western blotting method was used to determine the BMP-2 and Cbfal protein expressions. Results The ALP activities in LGTDG groups were lower than that in control group,especially in 1.0 mg·L-1 LGTDG group (P<0.01).The CCK-8 assay results showed that the proliferation rate of AS fibroblasts in 1.0 mg·L-1 LGTDG group was significantly low than that in control group(P<0.01),the proliferation rate reached the peak at the 4th day after LGTDG treatment and entered into the plateau phase,then the proliferation rate of the cells was decreased.The biochemical assay and Western blotting results indicated that the protein expression levels of BMP-2 in 1.0 mg·L-1 LGTDG group was significantly lower than that in control group (P<0.01).Conclusion LGTDG can effectively inhibit the BMP-2 expression in AS fibroblasts and delay the cells to differentiate into the osteoblasts and lead to AS ossification by BMP signal transduction pathway.