Therapeutic effect of Panax notoginseng saponins combined with cyclophosphamide in mice bearing hepatocellular carcinoma H22 cell xenograft / 南方医科大学学报

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.

Anti-tumor effects of Egr-IFNγ gene therapy combined with 125I-UdR radionuclide therapy / 中华放射医学与防护杂志

Objective To explore the anti-tumor effects of Egr-IFNγ gene therapy combined with 125I-UdR radionuclide therapy in mice bearing H22 hepatocarcinoma and its mechanism. Methods The recombinant plasmid pcDNAEgr-IFNγ mixed with liposome was injected into tumor. 48 h later, 370 kBq 125I-UdR was injected into tumor. The tumor growth rates at different times were observed. After 3 d gene-radionuclide therapy, the concentration of IFNγ in cytoplasm of H22 cells and cytotoxic activities of splenic CTL of the mice in different groups were examined. Results The tumor growth rates of pcDNAEgr-IFNγ +125 I-UdR group were obviously lower than those of control group, 125I-UdR group and pcDNAEgr-1 +125I-UdR group 6-15 d after gene-radionuclide therapy. IFNγ protein was found in cytoplasm of H22 cells in PcDNAEgr-1FNγ+125I-UdR group after 3 d gene-radionuclide therapy. Cytotoxic activity of splenic CTL in pcDNAEgr-IFN7 + 125I-UdR group was significantly higher than that in the other groups (P<0.01). Conclusions The anti-tumor effects in vivo of pcDNAEgr-IFNγ gene therapy combined with 125I-UdR radionuclide therapy are better than those of 125I-UdR therapy.

Matrine induced apoptosis of murine H22 hepatocarcinoma cells / 肿瘤

Objective: To investigate the apoptosis-inducing effect of matrine on murine hepatocarcinoma cell line H22 in vivo and in vitro and explore the underlying mechanisms. Methods: The H22 cell apoptosis induced by matrine at the early stage was detected with Annexin V-FITC/PI double staining assay, The expressions of Bcl-2 and Bax proteins in H22 cells as well as in the BALB/c H22 xenograft tumor tissues were detected using immunohistochemical method. Transmission electron microscopy (TEM) was used to observe the ultramicro-structure alterations of H22 xenograft tumor cells in BALB/c mice. The effect of matrine on the kinetics of tumor growth after subcutaneous injection of H22 cells in BALB/c mice was observed and the tumor inhibition rate was calculated. Results: Annexin V staining detected early apoptosis of H22 cells after treatment with matrine 1.0 mg/mL and 1.5 mg/mL for 48 h. The apoptotic rates were 11.71% and 17.86%, respectively, both of which higher than that of control groups (P <0.05). The tumor inhibition rate was above 60% after matrine treatment. Immunohistochemistry analysis revealed that matrine increased Bax protein expression and reduced Bcl-2 protein expression in both H22 cells in vitro and xenograft tumor tissues in vivo. TEM demonstrated the existence of apoptotic cells and apoptotic bodies in H22 xenograft tumor tissues after matrine treatment. Conclusion: Matrine significantly suppresses tumor growth and induced apoptosis both in vitro and in vivo. The apoptosis-inducing effects is related with up-regulation of Bax protein and down-regulation of Bcl-2 protein.

Study on activation of mouse peritoneal macrophage by Astragalus Saponin / 中国免疫学杂志

Objective:To investigate the immunomodulatory effect of Astragalus Saponin on macrophages and explore the mechanisms of its immunomodulation.Methods:By adding different concentrations of AS into cultured mice peritoneal macrophages,the influence of AS on synthesis of nitro oxide(NO)was observed by NO Kit(enzymic method).MTT assay was used to determine the cytotoxicity of macrophages induced by AS.The morphological changes of the macrophages were identified by Transmission Electron Microscope.LSCM and specificity fluorescent probe Fluo-3/AM were applied to observe the change of Ca2+ in the macrophages induced by AS.Results:AS could significantly increase NO synthesis,enhance the capacity of mice peritoneal macrophages for cytotoxicity to carcinoma cells.The surface projections of the macrophages were exhibited multiplicating,thickening and extenting via Transmission Electron Microscope.Augmented intracellular Ca2+ in the macrophage was observed by LSCM.Conclusion:AS can enhance the immune functions of macrophages,the increase of intracellular Ca2+ be one of the mechanisms of its immunomodulatory effects.

Activating effect of astragalus saponin on mouse peritoneal macrophages in vitro / 中国病理生理杂志

AIM:To investigate the immunomodulatory effect of astragalus saponin(AS)on macrophages and explore the mechanism of its immunomodulation.METHODS:By adding different concentrations of AS into cultured mouse peritoneal macrophages in vitro,the influence of AS on the synthesis of nitro oxide(NO)was detected by NO kit(enzymatic method).MTT assay was used to determine the cytotoxicity of macrophages induced by AS.The morphological changes of macrophages were observed under transmission electron microscope.LSCM and specificity fluorescent probe Fluo-3/AM were applied to observe the change of Ca2+ in macrophages induced by AS.RESULTS:AS significantly increased NO synthesis,enhanced the effects of mouse peritoneal macrophages on killing carcinoma cells.Cell surface projection exhibited multiplication,becoming thickening and growth longer via transmission electron microscope.Increase in intracellular Ca2+ in macrophages was also observed.CONCLUSION:AS enhances the immune function of macrophages by increasing NO synthesis and enhancing cytotoxicity.The increased intracellular Ca2+ may be the mechanism of immunomodulatory effect of AS.

Antitumor effect of H22 cells transfected with B7-1 in mice / 中国免疫学杂志

Objective:To study the effect of B7-1 costimulatory molecule in inducing tumor immunity.Methods:The lymphocytes were examined in vitro,for both proliferation induces(PI) and specific cytotoxic activity,H22 mice were challenged subcutaneously either by the mock following which the latency,survival time and tumor mass growth were noted.Results:The H22-B7-1 cells more effectively induced the proliferation of effecter lymphocytes and the generation of specific lytic activity against H22 cells.The H22-B7-1 cells demonstrated slower rate(P