| Objective:1. Analysis of the change about the distribution of MDSCs in liver, the proportion of MDSCs in spleen, the level of Arg-I in peripheral blood and the activity of NOS in serum of normal mice and different concentrations of5-fluorouracil (5-Fluorouracil,5-FU) groups.2. Research on the effects of5-FU on the proportion change, the distribution and the immune function of myeloid-derived suppressor cells (MDSCs) of hepatocarcinoma mice model, which provide experimental evidence for the clinical treatment of liver cancer.Methods:1. Using orthotopic transplantation of H22cell line method, the mice model of orthotopic liver cancer was established.2. On the7th day after transplantation, mice were randomly divided into four groups, each group included10mice. One group was injected with0.4ml PBS, and others were injected with doses (10,30and50mg/kg) of5-FU respectively for10days. Established normal control group at the same time.3. After finishing drugs injection, the distribution of MDSCs in liver tissues of mice was detected by immunohistochemistry; the proportion of MDSCs in spleen was tested by flow cytometry; the levels of IFN-y and Arg-I in serum were detected by enzyme-linked immunosorbent assay; the activity of NOS in serum was tested by chemical colorimetry; and the proliferation of T lymphocytes was detected by3-2,5-diphenyl-tetrazolium bromide (MTT) test.Results:1. With the increase of concentrations of the drug, the tumor volume reduced, and the adhesion of liver and abdominal cavity weakened.2. The level of MDSCs in liver of normal mice was low as (6.55±1.87)%. In PBS group,10mg/kg5-FU group,30mg/kg5-FU group, and50mg/kg5-FU group, the MDSCs levels were (24.58±3.79)%,(22.34±5.08)%,(16.03±3.25)%, and(10.71±2.68)%, respectively. The level of MDSCs in normal mouse spleen was low as (4.03±1.13)%, In the PBS group, 10mg/kg5-FU group,30mg/kg5-FU group, and50mg/kg5-FU group, the MDSCs levels were (16.97±3.08)%,(11.58±2.43)%,(8.88±2.05)%, and (6.29±1.69)%, respectively. The levels of MDSCs in liver and spleen were significantly higher than in the control group (P<0.05), and the highest proportion was in the PBS group. Increasing of concentrations of5-FU, the level of MDSCs declined gradually. Except for the PBS group and the10mg/kg group in liver, there was significant difference in other groups (P<0.05)3. The level of Arg-I in serum in normal mice was (30.37±1.24) ng/ml. In liver cancer mice model of the PBS group,10mg/kg5-FU group,30mg/kg5-FU group, and50mg/kg5-FU group, the level of Arg-I in serum were (136.28±6.77) ng/ml,(105.98±5.85) ng/ml,(72.77±6.36) ng/ml, and (53.66±8.86) ng/ml, respectively. The activity of NOS in serum of normal mouse was (20.59±4.58) u/ml. In liver cancer mice model of the PBS group,10mg/kg5-FU group,30mg/kg5-FU group, and50mg/kg5-FU group, the activity of NOS in serum were (57.22±8.49) u/ml,(46.77±6.39) u/ml,(37.60±8.28) u/ml,(27.17±4.91) u/ml, respectively. The level of Arg-I and the activity of NOS in the model were higher than the control group (P<0.05), and the PBS group showed the highest effect. As the increase of concentrations of5-FU, the level of Arg-I and the activity of NOS were declined gradually, in a dose dependent manner (P<0.05)4. The level of IFN-y in serum of normal mice was (147.45±36.12) pg/ml. In liver cancer mice model of PBS group,10mg/kg5-FU group,30mg/kg5-FU group, and50mg/kg5-FU group, the level of IFN-y in serum were (34.55±16.28) pg/ml,(29.90±8.76) pg/ml,(101.85±24.74) pg/ml, and (122.60±4.81) pg/ml, respectively. The level of IFN-γ in serum in cancer mice was lower than the control group, the lowest level was in the PBS group. Increasing of concentrations of5-FU, the level of IFN-y ascended gradually. Except for the PBS group and the10mg/kg group in serum, other groups had visible difference (P<0.05)5. The T cell proliferation assay showed that proliferation index in groups of spleen cells cultured for24h showed no significant difference. But at48h, the T cell proliferation index in normal mouse was3.23±0.73. In liver cancer mice model of the PBS group,10mg/kg5-FU group,30mg/kg5-FU group, and50mg/kg5-FU group, the proliferation index were1.26±0.18,1.30±0.17,2.02±0.72, and2.63±0.55, respectively. The cancer mice were lower than the normal control group (P<0.05), and the PBS group had the lowest proliferation. As the increase of concentrations of5-FU, the proliferation of T cells increased gradually. Except for the PBS group and the10mg/kg group, other groups showed significant change (P <0.05)6. The levels of MDSCs in liver and spleen were negatively correlated with the level of IFN-y and the proliferation of T cell at48h by the Pearson correlation analysis (P<0.01). The correlation coefficient were-0.843,-0.726,-0.784and-0.715, respectively. And the levels of MDSCs in liver and spleen were positively related to the levels of Arg-I and NOS (P<0.01). The correlation coefficient were0.873,0.798,0.909and0.845, respectively.Conclusion:5-FU, to a extent, could reduce the number of MDSCs in liver and spleen of liver cancer mice, relieve the immune inhibition function of liver cancer mice induced by MDSCs, and restrain the growth of the tumor, in a dose dependent manner. |
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