| With the development of immunology in recent years and research on the immune system,many diseases have a close relationship with low immunity.Therefore,research has found that a safe and effective immune enhancer is imminent.In this experiment,the immune enhancement of Kangai injection was studied from the molecular,cellular and overall levels.1.Effect on the immune activity of Raw 264.7 macrophageMethods: The experiment was divided into blank group,Kangai injection 40,80,120 ?L/mL group and positive control lipopolysaccharide 10 ?g/mL group.The blank group was given serum-free DMEM culture medium,the drug group was given Kangai injection 40,80 and 120 ?L/mL,the positive control group was given lipopolysaccharide 10 ?g/mL,and the pre-treatment for 24 h was used to detect the phagocytosis and adhesion ability of Raw 264.7 cells,intracellular ROS,NF-?B distribution,supernatant NO,TNF-? and IL-6 levels.Results: Kangai injection can significantly enhance the phagocytic ability and adhesion ability of Raw 264.7 cells(P<0.01),increase the levels of ROS,NO,TNF-? and IL-6(P<0.05 or P<0.01),and promote NF-?B nuclear translocation,suggesting that Kangai injection has the effect of enhancing the immune activity of Raw 264.7 cells.2.Effect of leukopenia on the number of peripheral blood leukocytes and the number of bone marrow nucleated cells in mice(1)cyclophosphamide-induced leukopenia modelMethods: The mice were randomly divided into blank group,model group,Kangai injection 5,10 and 20 mL/kg group and positive drug QLK 2 g/kg group.The model group,Kangai injection each dose group and the positive drug group mice were given intraperitoneal injection(ip)with cyclophosphamide 80 mg/kg on the first and third days of the experiment to establish a cyclophosphamide-induced leukopenia model.In the blank group and the model group,normal saline 20 mL/kg was given by ip,the drug group was given Kangai injection 5,10 and 20 mL/kg,and the positive drug group was given QLK 2 g/kg by intragastric administration(ig),once/days,a total of 7 days.Blood was taken from the tail vein on the 0th,2nd,4th,6th,and 8th day of the experiment,and the peripheral blood leukocyte count was performed.On the 8th day of the experiment,the femur was taken and the bone marrow nucleated cells were counted.Result: Kangai injection could significantly reduce the number of peripheral blood leukocytes and bone marrow nucleated cells in mice induced by cyclophosphamide(P<0.05 or P<0.01),suggesting that Kangai injection has obvious effect on leukopenia caused by cyclophosphamide.(2)X-ray induced leukopenia modelMethods: The mice were randomly divided into blank group,model group,Kangai injection 5,10 and 20 mL/kg group and positive drug QLK 2 g/kg group.The model group,Kangai injection each dose group and the positive drug group mice were given X-ray irradiation,0.305 GY/min,a total of 2.0 GY on the first day of the experiment to establish an X-ray induced leukopenia model.In the blank group and the model group,normal saline 20 mL/kg was given by ip,the drug group was given Kangai injection 5,10 and 20 mL/kg,and the positive drug group was given QLK 2 g/kg by intragastric administration(ig),once/ days,a total of 7 days.Blood was taken from the tail vein on the 0th,2nd,4th,6th,and 8th day of the experiment,and the peripheral blood leukocyte count was performed.On the 8th day of the experiment,the femur was taken and the bone marrow nucleated cells were counted.Results: Kangai injection can significantly reduce the number of peripheral blood leukocytes and bone marrow nucleated cells in mice induced by X-ray(P<0.05 or P<0.01),suggesting that Kangai injection has obvious resistance to X-ray-induced leukopenia effect.3.Effect on immune function of immunocompromised mice(1)cyclophosphamide-induced immunocompromised animal modelMethods: Mice were randomly divided into blank group,model group,Kangai injection 5,10 and 20 mL/kg group and positive drug levamisole 20 mg/kg group.Model group,Kangai injection each dose group and positive drug group mice were given phosphoramide 80 mg/kg by ip on day 1,3,and 5 to establish a cyclophosphamide-induced immunocompromised model.The blank group and the model group were given normal saline 20 mL/kg,the drug group was given Kangai injection 5,10 and 20 mL/kg,and the positive drug group was given levamisole 20 mg/kg once daily.day.The mouse thymus and spleen index,delayed type hypersensitivity,antibody production ability,macrophage phagocytosis ability,T lymphocyte transformation ability,NK cell activity and serum TNF-? and IL-6 levels were measured.Results: Kangai injection had no significant effect on the activity of NK cells in immunosuppressed mice induced by cyclophosphamide(P>0.05).Kangai injection can significantly improve the organ index,serum hemolysin,TNF-? and IL-6 levels of immunosuppressive mice induced by cyclophosphamide,enhance delayed type hypersensitivity,T lymphocyte transformation ability,macrophage phagocytosis ability(P<0.05 or P<0.01),suggesting that Kangai injection had a significant enhancement effect on cyclophosphamide-induced immunodeficiency in mice.(2)Hydrocortisone-induced immunocompromised animal modelMethods: Mice were randomly divided into blank group,model group,Kangai injection 5,10 and 20 mL/kg group and positive drug levamisole 20 mg/kg group.Model group,Kangai injection each dose group and positive drug group mice were given hydrocortisone 40 mg/kg by subcutaneous injection(sc)on the first,third,fifth and seventh day of the experiment to establish hydrocortisone-induced immunocompromised model.The blank group and the model group were given normal saline 20 mL/kg,the drug group was given Kangai injection 5,10 and 20 mL/kg,and the positive group was given levamizole 20 mg/kg once/day for 7 days.The mouse thymus and spleen index,delayed type hypersensitivity,antibody production ability,macrophage phagocytosis ability,T lymphocyte transformation ability,NK cell activity and serum TNF-? and IL-6 levels were measured.Results: Kangai injection can significantly improve the organ index,serum hemolysin,TNF-? and IL-6 levels in immunosuppressed mice,enhance delayed type hypersensitivity,macrophage phagocytosis,T lymphocyte transformation ability and NK cell activity(P<0.05 or P<0.01),suggesting that Kangai injection has a significant enhancement effect on hydrocortisone-induced immunocompromised mice.Conclusion:Kangai injection can enhance the immune activity of RAW 264.7 cells,increase the white blood cell count of leukopenia mice and the immunity of immunocompromised mice.It has a good immune enhancement effect and is expected to be a safe and effective immunopotentiator for clinical application. |
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