Effects Of Cinobufacini On Systemic Candidiasis In Mice

?Objective?To explore the effect and mechanism of cinobufacini on the anti-Candidal of the mouse peritoneal macrophages(M?),for approaching the effect of the anti-Candidal of cinobufacini,To investigate the effects of cinobufacini on systemic candidiasis in a murine model.and provide the experiment evidence of clinical anti-Candidal treatment.?Methods?The mouse peritoneal macrophage were incubated with cinobufacini and LPS in various concentrations for 24 h,then the treated macrophages were incubates with Candida albicans for 4h to determine the number of colony forming unit of C.albicans per milliliter and assay the concentrations of NO in the supernatant with Griess Reagent,and assay the concentrations of TNF-? in the supernatant with ELISA.We set cyclophosphamide-induced immuno-suppressed murine systemic candidiasis model,which was divided into two groups,control group(only candidiasis infection),and disposal group(candidiasis infection accompanying withcinobufacini injection).Colony forming units(CFU)of the kidneys and spleens were determined with plating dilution method.The histopathological changes and degree of infection of the kidneys and spleens were graded.Meanwhile,the levels of interferon gamma(IFN-?)in the spleens were measured by enzyme-linked immunosorbent assay.? Results ?1.With the cinobufacini concentration of 0.2mg/m L and 0.4mg/m L,the growth inhibition rates of candida were 35.6%and38.5% respectively,it had the significant difference(P<0.05)with the M? control group(the rate was 28.4%);2.With the concentration of 0.05?0.1?0.2 and 0.4mg/m L,cinobufacini can promote the production of TNF-?,there was significant difference concerned with the control group(P<0.05).3.And promoted the production of NO with the concentrations of 0.1 ? 0.2 ?0.4mg/m L of cinobufacini,it had the statistical significance concerned with the control group(P<0.05).In the control group,the CFUs of the 2d ? 3d ? 7d were 5.7865±0.1095 ?6.0970±0.0794? 5.9958±0.0817 respectively in kidneys and in spleens they were 5.3579±0.1174 ? 4.7599±0.1207 ? 3.6385±0.1851.Meanwhile,in the disposal group,the CFUs of the 2d?3d ? 7d were 4.9953±0.0726 ? 4.7751±0.1780 ? 3.9547±0.1234 respectively in kidneys and in spleens they were 5.2360±0.0696?4.6332±0.1139?3.5120±0.3120.The numbers of CFU of the fungi in the kidneys in the control group were significantly higher than those in the cinobufacini disposal group(P<0.01).The numbers of CFU of the fungi on the 2nd d,3rd d and 7th d after Candida albicans infection in the spleens in the control group were also higher than those in the disposal group,but without statistically significant difference(P>0.05).The scores of histopathological changes in the kidneys in disposal group were lower than those in the control group(P<0.05),and the degree of infection was milder in the disposal group.The scores of histopathological changes in the spleens in the disposal group were also lower than those in the control group,but without statistically significant difference(P>0.05).The levels of splenic IFN-? on the 2nd d,3 rd d and 7th d in the control group were 40.2389±4.1956 pg/ml? 59.3161±5.1913pg/ml?64.3723±5.1912 pg/ml respectively and in the disposal group they were 73.5312±5.9701pg/ml? 92.1207±7.5136 pg/ml?108.5212±9.2921 pg/ml respectively.The levels of splenic IFN-? in the disposal group were significantly higher than those in the control group(P<0.01).?Conclusion?Cinobufacini can promote the anti-candidal activeeity of the macrophages obviously,it may be concerned with the inhibition of the secretion of TNF-? and NO by macrophages,It is demonstrated that cinobufacini has protective effect on murine systemic candidiasis.