| Objectives To investigate the effect of formononetin on immunity function of mice treated with different doses of formononetin,so to provide theoretical basis for clinical application of formononetin.Methods A total of 40 SPF grade kunming male mice were randomly divided into four groups as the followings,low,medium and high dose formononetin experimental groups and control group,10 mice in each group.Mice in the above 3 experimental groups were injected intramuscular with 0.1 ml of formononetin solution at 10mg/kg/d,20mg/kg/d and30mg/kg/d respectively,while mice in the control group were treated with equal volume of normal saline.After 4 weeks of continuous injection,tail blood of mice was collected in a sterile way,and the abilities of neutrophils of mice to phagocyte staphylococcus in each group were compared.Mice were intraperitoneally injected with1 ml of chicken erythrocyte suspension with a concentration of 10%,thereafter the phagocytic functions of macrophages in each group were detected.Blood was collected from tail vein,following that serum was isolated.The contents of lysozyme and Ig G in serum of mice were determined by one-way agar diffusion assay.The concentrations of IFN-γ and IL-4 in serum were determined by enzyme immunoassay.Mice were killed with ether,Spleen cells were isolated.The apoptosis rate of spleen cells were determined by TUNEL assay and transformation rate were conducted by morphological method.Results 1 Effect of formononetin on phagocytic cell function in mice.The neutrophil phagocytosis rate of mice in low dose,middle dose and high dose group and control group were 80.12±2.63%,81.97±3.11,83.61±2.17% and 76.19±4.73%.Statistic analysis demonstrated that the neutrophil phagocytosis rate of mice in different doses of formononetin were higher than that in control group,The differences were statistically significant(P<0.05).However,there were no significant differences regarding neutrophil phagocytosis rate among the 3 formononetin groups(P>0.05);The phagocytic rates of macrophages in the low-dose,medium-dose,high-dose groups and the control group were 38.81±1.53%,41.80±1.48%,42.53±2.09% and 35.95±2.77%respectively.Statistic analysis discovered that the phagocytic rate of macrophages in the medium and high dose groups were higher than those in the control group and the low dose group,and the difference was significant(P<0.05).However,the phagocytic rate of macrophages in the low dose formononetin group was not significantly different with that in the control group(P>0.05),and the similar result was observed in medium dose group and high dose group.2 Effect of formononetin on lysozyme content in serum of mice.The lysozyme contents in low dose,medium dose,high dose groups and control group were16.38 ±0.15 IU/ml,23.62±0.68 IU/ml,25.16±0.52 IU/ml and 17.91±0.26 IU/ml respectively.Statistic analysis showed that the serum lysozyme contents in medium and high dose formononetin group were higher than those in control group and low dose group,and differences were significant(P<0.05).Compared to the control group,the content of serum lysozyme in low dose formononetin group showed no significant difference(P>0.05).Serum lysozyme content in medium dose group was not significantly different with that in high dose group(P>0.05).3 Effects of formononetin on serum Ig G,IL-4 and IFN-γin mice.The contents of Ig G in serum of low dose,medium dose,high dose group and control group were 5.93±0.57mg/ml,7.17±0.68mg/ml,7.26±0.53mg/ml and 5.21±0.39mg/ml,respectively.Statistic analysis showed that the contents of Ig G in serum of mice in medium and high dose formononetin groups were higher than those in control group and low dose groups,and the differences were significant(P<0.05).However,there was no significant difference regarding Ig G content between low dose and control group,so was that of medium dose and high dose group(P>0.05).The IL-4 content in serum in the low,medium and high dose groups and control group were 91.96±9.99pg/ml,107.74±7.47pg/ml,112.59±8.41pg/ml and 80.95±8.33pg/ml,respectively.Statistic analysis showed that the contents of IL-4 in serum of mice in the low,medium and high dose formononetin groups were higher than that in control group,and the difference were significant(P<0.05);The IL-4 contents in serum of mice in medium and high dose formononetin groups were higher than that in low dose group,and the difference were significant(P<0.05),while there was no significant difference between the medium dose and high dose groups(P>0.05).The levels of IFN-γ in the serum of mice in the low,medium and high dose groups and the control group were 98.66±13.54pg/ml,105.48±17.26pg/ml,96.36±15.55pg/ml and 103.48±9.22pg/ml,respectively.Statistical analysis displayed that there was no significant difference in the content of IFN-γ in serum between the 4 groups(P>0.05).4 Effects of formononetin on apoptosis rate and transformation of spleen cells in mice.The apoptosis rates of spleen cells in low,medium and high dose groups and the control group were 7.16±2.38%,4.29±2.62%,4.38±2.52%and 12.38±6.19%,respectively.Statistical analysis displayed the apoptosis rate of spleen cells in the low dose,medium dose and high dose formononetin group were lower than that in the control group(P>0.05);The apoptosis rate of spleen cells in medium and high dose groups were lower than that in low dose group(P>0.05).The lymphocyte transformation rates of the low,medium and high dose groups and the control group were 36.02±2.20%,40.41±2.47%,43.27±3.71% and 33.91±3.27%,respectively.Statistical analysis displayed the spleen cell transformation rate of mice in the medium and high dose groups were higher than those in low dose group and the control group,and the difference was significant(P<0.05);However,there was no significant difference between low dose group and control group,so was between medium dose group and high dose group(P>0.05).Conclusions1 Formononetin was likely to improve the phagocytosis of innate immune cell and the antibacterial activity of lysozyme in mice.2 Formononetin was seen to enhance the adaptive humoral immunity of mice by inhibiting the apoptosis of spleen lymphocytes and increasing the transformation rate in a manner of dose-dependent,yet it has even little effect on the adaptive cellular immunity of mice.Figure 7;Table 4;Reference 91... |
