ABSTRACT
Objective To observe the effect of heat shock protein 70 (HSP70) expression induced by Herba Hedyotis Diffusae (HHD) on the apoptosis of H22 hepatoma cells.Methlds Mice were immunized with H22 hepatoma cells from in-vivo ascites of the mice pretreated by HHD and Radix Codonopsis (RC).Mice were divided into five groups after being blocked with HSP70 monoclonal antibody:non-HHD blocking group,HHD blocking group,non-RC blocking group,RC blocking group and the blank control group,10 mice in each groups.Then the mice were attacked with cultured H22 hepatoma cells to induce transplanted tumor.The apoptosis in the transplanted tumor was detected by a terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling(TUNEL) method.Results Large amount of brown particles can be observed in the cell nucleus of transplanted tumor mice from HHD groups.Compared with RC groups,number of apoptotic cells and the staining intensity were increased in HHD groups.The apoptosis in HHD groups and RC groups differed from that in the blank control group,and was obvious in HHD groups.The apoptosis exerted a increasing tend in non-HHD blocking group,but the difference was insignificant between non-HHD blocking group and HHD blocking group,and between non-RC blocking group and RC blocking group.Conclusion HHD can promote the apoptosis of H22 hepatoma cells in transplated tumor through inducing the expression of HSP70 to some extent,and the related mechanism of inducing apoptosis needs further research.
ABSTRACT
【Objective】To observe the effect of Herba Hedyotis Diffusae(HHD)on the histological features of living murine ascites hepatoma H22 cells as well as CD+4 and CD+8 expression in T lymphocytes.【Methods】Special pathogen free(SPF)Kunming mice were randomized into 4 groups:model group,HHD(25 g/kg)group,Radix Codonopsis(RC,25 g/kg)group,heat-shock group.The murine ascites hepatoma H22 model was established by abdominal transplantation of H22 cells.The histological features of H22 cells from living murine ascites were examined through living mice ascites smear and giemsa staining.The CD+4 and CD+8 expression in ascites T lymphocytes was detected by immunohistochemical method.【Results】H22 cell deformation such as cell membrane incomplete and vesicant,some nucleus being migration and pycnosis was obvious in HHD and heat-shock groups.The expression of both CD+4 and CD+8 expression in T lymphocytes of HHD groups was increased,but only CD+4 expression in RC group and only CD+8 expression in heat-shock group was increased.【Conclusion】HHD exerts some inhibitory-killing effect on living murine H22 cells,and its mechanism may be related to the increase of immune function and to the cytotoxic effect.
ABSTRACT
[Objective] To compare the effects of heat-clearing and detoxifying herbs with that of tonifying herbs on mice hepatoma cells line H22, and to explore their immunologic mechanisms. [Methods] Herba Rabdosiae Rubescentis (HRR) and Herba Scutellariae Barbatae (HSB) were selected as the heat-clearing and detoxifying herbs, and Radix Astragali (RA) as the representative of tonifying herbs. Fifty KM mice were inoculated with hepatoma cells line H22 and then were randomized into five groups: HRR group (30 g?kg-1?d-1 by gavage for 12 days), HSB group (15 g?kg-1?d-1 by gavage for 12 days), RA group (15 g?kg-1?d-1 by gavage for 12 days) , heat-shock group (treated 20 min at 43℃, once every 4 days and three times in all) and model group. Twelve days later, the mice were executed and the ascites smears were prepared. The cell feature in ascites was observed under light microscope and the expression of CD4+ and CD8+ on the cellular membrane of the T lymphocyte in ascites was detected with immunohistochemical method. [Results] A large amount of damaged and apoptotic H22 cells were found in HRR, HSB and heat-shock groups, and the changes of cell feature in RA group were not obvious. The expressions of CD4+ and CD8+ on the membrane of T cells in ascites was increased in HRR and HRB groups, but the expression of CD4+ was increased in RA group, and CD8+ expression was increased in heat-shock group.[Conclusion] Heat-clearing and detoxifying herbs can induce the necrosis and apoptosis of mice hepatoma cells line H22 and the mechanism may be related to the increase of cellular immunity, thus enhancing the body immunity to kill the tumor cells. RA can not kill the tumor cells directly and its antitumor action may be related to the increase of CD4+ expression.
ABSTRACT
Objective To observe the effect of Herba Hedyotis Diffusae(HHD) on expression of heat shock protein 70(HSP70) and P16 anti-oncogene protein in mice with H22 cell transplanted tumor. Methods The mice were immunized with hepatoma-lines-H22 cells which were treated by Chinese herbs firstly. According to the pretreatment of the Chinese herbs and whether the H22 cells being blocked by HSP70 monoclonal antibody,the mice were divided into 5 groups:HHD non-blocking group,HHD-blocking group,Radix Codonopsis(RC)-blocking group,RC non-blocking group,and RPMI-1640 control group. Then the treated H22 Cells were translated into the mice. The expression of HSP70 and P16 anti-oncogene protein in mice with transplanted H22 liver carcinoma was detected by immunohistochemical marking method. Results The expressions of HSP70 was up-regulated in HDD non-blocking group and RC non-blocking group,in particular in RC non-blocking group,and the difference was significant as compared with HHD-blocking group,RC-blocking group and control blank group.The expression of P16 anti-oncogene protein was up-regulated in the treatment groups,and the difference was significant in comparison with the control group. The effect on P16 anti-oncogene protein expression was stronger in HDD groups than that in RC groups,but the difference was insignificant between the blocking group and the non-blocking group of each herb. Conclusion The anti-tumor effect of HDD may be achieved by inducing HSP70 expression in transplanted H22 neoplasia and by enhancing the immunogenicity. The up-regulation of P16 anti-oncogene expression by HHD and RC has no direct relationship with the induction of HSP70,and the related mechanism needs further research.