Study On Toxicity Attenuation And Antitumor Synergy Of The Secondary Metabolites Of Rhizotonia Legminicola On Adriamycin

Subject: Established the model of adriamycin-induced myocardium injury and bone marrow arrest in mice to investigate the protective effects of the secondary metabolites of rhizoctonia leguminicola on adriamycin(ADR)toxicity. It studied the protective effects of the secondary metabolites of rhizoctonia leguminicola by blood routine analysis,ECG test,biochemical index mensuration,pathology changes observation and so on. It also investigated the co-antitumor effects of the secondary metabolites of rhizoctonia leguminicola and adriamycin. All these could provide new thoughtfulness of the application of the secondary metabolites in antitumor treatment.Method:①the mice were injected intraperitoneally by 2mg·kg-1,4mg·kg-1,6mg·kg-1 ADR respectively every other day for 5 times, and some mice were injected by 20mg·kg-1 ADR one time. By Blood routine analysis,biochemical index mensuration,pathology changes observation, it analysed the injury degree of ADR in mice to establish the feasible way of adriamycin-induced myocardium injury and bone marrow arrest.②The mice were infused the secondary metabolites of rhizoctonia leguminicola which had swainsonine and injected ADR after 6h, the doses of swainsonine were 8mg·kg-1,16mg·kg-1and 32mg·kg-1. 24h after the last administration,blood routine and ECG were tested, the activities of creatine kinase (CK) and lactate dehydrogenase(LDH)in serum, superoxide dismutase(SOD) and the content of malondi- aldehyde(MDA) in myocardium were examined. Pathological changes were observed in myocardium. It would Investigate the protective effects and protective dose of the secondary metabolites of rhizoctonia leguminicola to the adriamycin-induced myocardium injury and bone marrow arrest in mice.③Cancer model were estabilished by transplanting S180 cells to mice. The mice were divided into 4 groups randomly: control group(Ⅰ,normal saline),SW group(Ⅱ, SW16 mg·kg-1), ADR group(Ⅲ,ADR2.5mg·kg-1),combination group(Ⅳ,SW16 mg·kg-1+ADR2.5mg·kg-1). 48h after the last administration, the proliferation index of T lymphocytes and the phagocyte ability of peritoneal macrophages were determined. It also calculated the rate of tumor inhibition and spleen index,and observed the pathology changes by optics microscope.④After being treated with the secondary metabolites of rhizoctonia leguminicola at different dose(10,5,2.5,1.25,0.63,0.32μg·ml-1)for 72 h,the IC50 of this secondary metabolites to HepG2 cells were examined. The survive rate of HepG2 cells were examined after being treated with this secondary metabolites and ADR(60μg·ml-1,6μg·ml-1) for 72h.Result:①Injecting ADR intraperitoneally at a dose of 4mg·kg-1 every other day for 5 times could induce bone marrow arrest and myocardium injury in mice.②The secondary metabolites of Rhizoctonia leguminicola could attenuates adriamycin-induced myocardium injury and bone marrow arrest in mice. It prevented leukocyte count, erythrocyte count and haematoglobin content from decreasing. It increased heart rate, decreased Q-wave amplitude, shortened QRS duration and rose S-T segment. It also decreased the activities of serum CK and LDH, content of myocardial MDA, increased the activity of myocardial SOD. Moreover, it preserved the mouse myocardial morphology.③The tumor inhibitory rate was highest(64%)in groupⅣin vivo, and it indicated that the secondary metabolites of rhizoctonia leguminicola could enhance the antitumor effects of ADR. The spleen index was 38.97±4.03 in groupⅣ,and it was higher than groupⅢ(p<0.01). The proliferation index of T lymphocytes was 1.22±0.03 in groupⅣ, it was also higher than groupⅢ(p<0.05). There was large area putrescence in tumor tissue in groupⅣ. The result showed that the secondary metabolites could attack tumor cells directly and stimulate immune responses to inhibit tumor growth.④with the concentration of the secondary metabolites increased, the survive rate of HepG2 cells decreased,and the IC50 in 72h was1.25μg·ml-1. While the secondary metabolites was combined with ADR to HepG2,the survive rate had no change, it might be that the immunostimulation effects played an important role in tumor inhibition.Conclusion:①Injecting ADR intraperitoneally at a dose of 4mg·kg-1 every other day for 5 times could induce bone marrow arrest and myocardium injury in mice.②The secondary metabolites of Rhizoctonia leguminicola (16mg·kg-1,32mg·kg-1SW)could attenuates adriamycin-induced myocardium injury and bone marrow arrest in mice.③The secondary metabolites of rhizoctonia leguminicola could enhance the antitumor effects of ADR on transplanting S180 rather than HepG2 cells.