Effect Of HIIT Pretreatment On NK Cell Function In Melanoma Mice

Research purpose:Current studies have focused on the enhancement of NK cell function after exercise intervention in tumor-bearing mice to exert the anti-tumor function of NK cells,but the effects and mechanisms of exercise intervention at different stages of melanoma development are unclear.In this study,we investigated the effects of HIIT pretreatment on NK cell development and NK cell metabolic function in melanoma mice to reveal the effects of HIIT pretreatment on the anti-tumor effect of NK cells,and thus provide new ideas for exercise-assisted clinical treatment.Research methods:Fifty-seven SPF-grade 6-week-old female C57 mice were housed for one week in an SPF-grade animal room before the experiments.Experiment 1:divided into 2 groups,tumor group(T group,N=9)and tumor motility group(THs group,N=9).no motility in the T group after subcutaneous injection of 100 μL melanoma cell suspension;a 2-week HIIT intervention was started 24 hours after subcutaneous injection of 100 μL melanoma cell suspension in the THs group.Experiment 2:divided into three groups:normal control group(group C,N=13),short-term exercise tumor group(group TH,N=13)and long-term exercise tumor group(group HTH,N=13).group C was injected intraperitoneally with 100μL of PBS buffer;group TH was injected intraperitoneally with 100μL of melanoma The HTH group was injected intraperitoneally with 100μL of melanoma cell suspension after eight weeks of exercise intervention and continued with exercise intervention for two weeks after 24 hours of injection.The maximum exercise capacity was tested before exercise intervention,four weeks after,and before tumor injection.Mouse spleen and tumor tissues were taken,prepared as single-cell suspensions,and incubated with antibodies.The percentage of NK cells and their subtypes,NK cell checkpoints,the mitochondrial mass of NK cell subtype cells,glucose uptake capacity,and ROS expression in each tissue were measured by flow cytometry.Research results:1.Survival time showed that mice in the THs group injected with melanoma cells followed by 2 weeks of exercise intervention had a higher survival time than those in the T group without exercise intervention,and the survival times of the two groups were statistically different2.The maximal locomotor ability test showed that after 8 weeks of HIIT intervention,the maximal locomotor speed of mice in the HTH group was significantly higher than that of mice in the C and TH groups,and the maximal locomotor speed of mice in the HTH group was significantly higher compared to that before the locomotor intervention.3.NK cell results showed that:(1)The NK cell percentage results showed that in the tumor tissue,the percentage of NK cells in the HTH group was significantly higher than that in the TH group.(2)The results of NK cell subtype percentage showed that in the spleen tissue,the percentage of DN cells in the HTH group was statistically lower than that in the TH group;the percentage of CD27SP cells in the HTH group was significantly lower than that in the TH group;the percentage of CD27SP cells in the TH The percentage of CD27SP cells was higher in the TH group than in the C group,with a significant difference.the DP cells in the HTH group were significantly higher than those in the TH and C groups.4.Glucose uptake capacity of different subtypes of NK cells:in splenic tissue,the percentage of glucose uptake by DN cells was significantly lower in the HTH group than in the TH group.5.Mitochondrial mass of different cell subtypes of NK cells.(1)Changes in the spleen:the percentage of mitochondrial mass of CD27SP cells was lower in the HTH group than in the C group and was statistically significant.(2)Changes in tumors:the mean mitochondrial fluorescence intensity of CD27SP cells in the HTH group was significantly lower than that in the TH group.the mean mitochondrial fluorescence intensity of DP cells in the HTH group was statistically higher than that in the TH group.the mitochondrial activity of CD11bSP cells in the HTH group was statistically higher than that in the TH group.Research conclusion:HIIT pretreatment promotes the infiltration of mature NK cells in tumor tissues and improves the aerobic metabolic capacity of mature NK cells to enhance the anti-tumor effect of the organism.