Secretory Function of Kupffer Cells in Liver Cancer After Cryoablation

Objective To explore the effect of Kupffer cells (KCs) secretion on HCC cells after cryoablation and its mechanism.

Methods Sixteen groups were divided:37℃ control group, three hypothermia groups(0℃, 5℃, 10℃), freeze-thaw necrotic substances group, three combined stimulation group and 8 parallel groups for each group. In the 8 parallel groups, pyrrolidine dithiocarbamate(PDTC) was added, and the final concentration was 100 μmol/ml. The concentration of TNF-α, IL-1β and INF-γ in 16 groups of KCs supernatant were determined by Elisa method. The expression of NF-κB protein in each group was detected by Western blot.

Results The secretion of inflammatory factors TNF-α, IL-1β and INF-γ increased after combined stimulation of KCs with hypothermia and freeze-thaw necrotic substances (P<0.01). The combined stimulation of low temperature (0℃, 5℃, 10℃) and freeze-thaw necrotic substances in the three groups showed superposition effect. Hypothermia stimulation had no significant effect on the expression of NF-κB protein (P>0.05), but the expression of NF-κB protein in the combined stimulation group and freezethaw necrosis substances group were significantly up-regulated (P<0.05). Treated with NF-κB inhibitor, the expression of NF-κB protein and the secretion level of inflammatory factors did not change significantly among 8 parallel groups (P>0.05). The concentrations of TNF-α, IL-1β and INF-γ were positively correlated with the expression of NF-κB protein in the culture medium of KCs (r ≥ 0.870, P=0.000).

Conclusion Freeze-thaw therapy could enhance the function of KCs secretion cytokines, and to a certain extent, it could induce inflammatory response and eliminate tumor cells. The secretion function of KCs may play a role through NF-κB signaling pathway.