Effect of Phyllanthus emblica L.total flavonoids on immunomodulatory function in H22 tumor-bearing mice
OBJECTIVE To explore the effect of Phyllanthus emblica L.(PEL)total flavonoids on immunomodulatory function in H22 tumor-bearing mice and examine its relationship with changes in tumor microenvironment.METHODS After an inoculation of H22 cell,50 Kunming mice were randomized into five groups of Control,5-fluorouracil(5-FU)and low(YGZY-L:0.496 g·kg-1),medium(YGZY-M:0.991 g·kg-1)and high(YGZY-H:1.982 g·kg1)doses of PEL total flavonoids(n=10 each).Tumor weight and tumor inhibition rate(TIF)were recorded after 14-day dosing.The levels of interleukin-2/6/10(IL-2/6/10),interferon-gamma(IFN-γ)and tumor necrosis factor-alpha(TNF-α)in tumor tissues were detected by enzyme-linked immunosorbent assay(ELISA).Hematoxylin-eosin(HE)stain was utilized for observing the morphology of tumor tissues and immunohistochemistry for detecting the ratio of CD4+T/CD8+T cell in tumor tissues.Single-cell suspension of tumor tissues was prepared for isolating lymphocytes and the proportion of infiltrating lymphocytes in tumor tissues was observed by flow cytom-etry.Western blot was utilized for observe the expressions of NF-κB1,FLT3,PD-1,BTLA and CTLA-4.RESULTS TIF was 62.2％in 5-FU group,13.3％in YGZY-L group,40.7％in YGZY-M group and 53.5％in YGZY-H group.HE stain indi-cated that PEL total flavonoids induced cellular necrosis and nuclear condensation.Immunohistochemistry revealed that PEL total flavonoids and 5-FU could activate the number of CD4+T/CD8+T cells in tumor microenvironment(P＜0.01).PEL total flavo-noids significantly up-regulated the levels of IL-2,IL-6,IFN-y and TNF-α and down-regulated IL-10 in tumor microenviron-ment(P＜0.01).The proportions of CD4+CD25+Foxp3+Treg,CD4+PD-1+TIL and CD8+PD-1+TIL dropped while the pro-portion of CD8+CD28+CTL rose(P＜0.05 or P＜0.01).As compared with Control group,PEL total flavonoids down-regulated markedly the expressions of NF-κB1/FLT3(P＜0.01)and PD-1/CTLA-4/BTLA(P＜0.05 or P＜0.01)in tumor microenvironment.CONCLUSION PEL total flavonoids may treat liver cancer tumors through improving tumor immune micro-environment and modulating immune function of tumor-bearing mice.
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