Effect and mechanism of formononetin mediated TLR4/iNOS pathway on local immune regu-lation in tumor bearing mouse model of pancreatic cancer
Objective To investigate the effect of formononetin mediated Toll-like receptor 4(TLR4)/inducible nitric oxide synthase(iNOS)pathway on local immune regulation in tumor bearing mouse model of pancreatic cancer.Methods 50 BALB/c mice were injec-ted with mouse pancreatic cancer cells MPC-83 under the armpit to establish a tumor bearing mouse model of pancreatic cancer.The successfully modeled mice were randomly divided into model group,gemcitabine group(0.5 mg/g),low,medium and high dose form-ononetin groups(25,50 and 100 g/kg),with 10 in each group.3 days after the end of administration,the tumor inhibition rate was cal-culated,and hematoxylin eosin(HE)staining was used to observe the micropathology of tumor tissue,and the content of NO was deter-mined by Griess method,and the expressions of TLR4,nuclear transcription factor-κB p65(NF-κB p65)and iNOS mRNA were de-tected by real-time quantitative polymerase chain reaction,and the expressions of TLR4,NF-κB p65,iNOS protein and the level of p-NF-κB p65 were detected by western blotting.Results Compared with model group,tumor volume in gemcitabine group and form-ononetin 3 dose groups were decreased on day 7,14,21 and 28,and tumor mass,the content of NO,and the mRNA and protein expres-sions of TLR4,NF-κB p65,iNOS and the level of p-NF-κB p65 in tumor tissue were significantly decreased,with statistically sig-nificant(P＜0.05 or P＜0.01).Conclusion Formononetin can improve the local immune function of pancreatic cancer bearing mice,which may be related to the inhibition of TLR4/iNOS pathway in tumor microenvironment,the down-regulation of NO content.
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