ABSTRACT
OBJECTIVE@#To investigate the therapeutic effects of total saponins from Panax notognseng (PNS) combined with cyclophosphamide (CTX) in mice bearing hepatocellular carcinoma H22 cell xenograft.@*METHODS@#We examined the effects of treatment with different concentrations of PNS on H22 cell proliferation for 24 to 72 h in vitro using CCK8 colorimetric assay. Annexin V/PI double fluorescence staining was used to detect the effect of PNS on apoptosis of H22 cells. Mouse models bearing H22 cell xenograft were established and treated with CTX (25 mg/kg), PNS (120, 240 or 480 mg/kg), alone or in combinations. After treatments for consecutive 10 days, the mice were euthanized for examinations of carbon clearance ability of the monocytes and macrophages, splenic lymphocyte proliferation, tumor necrosis factor (TNF-α), interleukin-2 (IL-2), serum hemolysin antibody level, blood indicators, and the tumor inhibition rate.@*RESULTS@#Treatment with PNS concentration-dependently inhibited the proliferation and significantly promoted apoptosis of cultured H22 cells (P < 0.01). In the tumor-bearing mouse models, PNS alone and its combination with CTX both resulted in obvious enhancement of phagocytosis of the monocyte-macrophages, stimulated the proliferation of splenic lymphocytes, promoted the release of TNF-α and IL-2 and the production of serum hemolysin antibody, and increased the number of white blood cells, red blood cells and lymphocytes in the peripheral blood. Treatment with 480 mg/kg PNS combined with CTX resulted in a tumor inhibition rate of 83.28% (P < 0.01) and a life prolonging rate of 131.25% in the mouse models (P < 0.05).@*CONCLUSION@#PNS alone or in combination with CTX can improve the immunity and tumor inhibition rate and prolong the survival time of H22 tumor-bearing mice.
Subject(s)
Animals , Humans , Mice , Carcinoma, Hepatocellular/pathology , Cyclophosphamide/therapeutic use , Hemolysin Proteins , Heterografts , Interleukin-2 , Liver Neoplasms/pathology , Panax notoginseng , Saponins/therapeutic use , Tumor Necrosis Factor-alphaABSTRACT
OBJECTIVE:To investigate the effect of KCa3.1 channel on the function of EPCs. METHODS: The gene expression of KCa3.1, vWF and CD31 on EPCs were detected by qRT-PCR. CCK-8 kit, cell adherent method and Matrigel were used to detect the changes of cell proliferation, adhesion and in vitro angiogenesis; cell immunofluorescence or fluorescence activated cell sorter(FACS) was used to detect the protein expression of KCa3.1, vWF, CD31, integrin β1, integrin β3 separately. RESULTS: Blocking the function of KCa3.1 or interfering with the expression of KCa3.1 can attenuated EPC function of proliferation, adhesion and angiogenesis, but it can promote the differentiation of EPCs. Overexpression or activation of KCa3.1 channel can enhance EPCs proliferation, adhesion and angiogenesis but decrease the level of differentiation. The expression of integrin β1 on EPCs was attenuated with blocking KCa3.1 channel, but the expression effect was reversible by the activator. CONCLUSION: The alteration of KCa3.1 channel function or expression affect the biological characteristics and differentiation of EPCs.
ABSTRACT
OBJECTIVE@#To investigate the effects of endothelial progenitor cells (EPCs) under shear stress on the biological function such as proliferation, adhesion, migration, apoptosis and expression of α-smooth muscle actin (α-SMA), collagen-I and collagen-Ⅲ of hepatic stellate cells (HSCs).@*METHODS@#HSCs and EPCs were inoculated into the upper and lower layers of the co-culture chamber respectively and co-incubated for 24 hours. Then, 12 dyne/cm shear stress was applied to EPCs cells for another 24 hours. After that, proliferation, adhesion, migration and apoptosis of HSCs were detected by cell counting kit-8 (CCK-8) kit, cell adherent assay, Boyden cell migration assay and flow cytometry respectively. Fluorescence quantitative PCR and Western blot were used to detect the mRNA and protein expression of alpha -SMA, collagen I and collagen-Ⅲ in HSCs.@*RESULTS@#Under shear stress, EPCs ecological niche could obviously inhibit the proliferation, adhesion and migration of HSCs, promote the apoptosis of HSCs, and down-regulate the mRNA and protein expression of collagen-I, collagen-Ⅲ in HSC cells.@*CONCLUSIONS@#Under shear stress, EPCs ecological niche could inhibit the fibrosis development of HSCs to a certain extent.