| Background:Malaria is an insect-borne infectious disease caused by Plasmodium parasites bitten by anopheles mosquitoes.It is a serious threat to human life and has attracted worldwide attention.Artemisinin-based combination therapies(ACTS)have played an indispensable role in the treatment of malaria patients.However,the emergence of artemisinin-resistant strains poses a major new challenge to malaria cure and an urgent need to find new antimalarial drugs.Zincpolyanemine(ZPT)was the first zinc compound containing mercaptopyridine extracted from the root of the annonaceae plant Lingshishuomia,which has antibacterial,antimalarial and antitumor effects.Objective:In this paper,artemisinin-resistant and chloroquine-resistant Plasmodium bergi K173strain(Pb K173)was selected to establish artemisinin-resistant and chloroquineresistant mice malaria model.The combination of Artesunate(ART)with different doses of ZPT and Benflumetol with different doses of ZPT was investigated.In order to provide a basis for ZPT in Lingshuiangluo as a new antimalarial drug,and provide a new idea for drug combination treatment of malaria.Methods:1.The 65 th genemiceion of artemisinin-resistant P.berghei mouse malaria model was established by artemisinin-resistant P.berghei strain K173(Pb K173);Model verification: Model mice were given different doses of artemisinin by gavage for 4days.The general situation of mice was observed,and the average infection micee and resistance index of protozoa were detected by daily weighing and blood smear.2.The acute toxicity of ZPT was investigated by single intragastric administmiceion of ZPT.The appearance,mental state,behavior,fur,breathing,urine and feces,color changes,diarrhea,death,and abnormal secretions in eyes and nose of the mice were observed every day for 14 days after administmiceion.3.In the 65 th genemiceion of artemisinin-resistant Plasmodium Birkeni model,different doses of ZPT,ART and their combination were given,and the infection micee and relapse micee of mice were observed by blood smear.The expression levels of inflammatory factors TNF-α,IL-6 and anti-inflammatory cytokine IL-10 in serum were detected by ELISA kit,and the coefficients of liver and spleen organs were calculated.To compare the efficacy of ZPT,ART alone or combined against artemisinin-resistant murine malaria.4.The chloroquine-resistant mouse malaria model was established,and different doses of ZPT,BEN and their combination were given respectively.The infection micee and relapse micee of mice were observed by blood coating,the expression levels of inflammatory factors TNF-α,IL-6 and IL-10 in serum were detected by ELISA kit,and the coefficients of liver and spleen organs were calculated to investigate the pathological changes of liver and spleen tissues.To compare the efficacy of ZPT,BEN alone or combined against chloroquine-resistant murine malaria.Results:1.24 h after administmiceion,the average infection micee of the protozoa in the(1)to(5)groups of mice was more than 0.5%,and the 65 th genemiceion of artemisininresistant Plasmodium brucei strains showed obvious resistance to artemisinin.The RI was: 16.37,indicating successful modeling,which could be used as a model for later experiments.2.Compared with the control group,the mice were given ZPT 200 mg·kg-1 by intragastatic administmiceion,and no obvious poisoning such as mental distress,limb weakness,or death occurred.There were no significant differences in liver and spleen coefficients(P > 0.05),no obvious abnormalities in tissue morphology and pathology of liver and spleen(P > 0.05),no significant differences in body weight within 14days(P > 0.05).3.24 h after the last administmiceion,there was no significant difference in the average infection micee among the experimental groups(P > 0.05).On the 7th and14 th day after drug withdrawal,compared with the control group,ART group,ZPT group,ZPT+ART group,2x ZPT+ART group,2.5x ZPT+ART group,The average infection micee of mice in 3x ZPT+ART group was significantly decreased(P < 0.05).Compared with ART group,the average infection micee of mice in ZPT+ART group,2x ZPT+ART group,2.5 ZPT+ART group and 3x ZPT+ART group was significantly decreased(P < 0.05).The average inhibitory micee was significantly different among all groups(P < 0.05).Compared with the control group,the liver and spleen coefficients of mice in ART group,ZPT group,ZPT+ART group,2x ZPT+ART group,2x ZPT+ART group,3x ZPT+ART group and 3x ZPT+ART group were decreased,and the liver and spleen coefficients of mice in 2x ZPT+ART group and 3x ZPT+ART group were decreased significantly(P < 0.05);Compared with the control group,the serum contents of TNF-α,IL-6 and IL-10 in each group were decreased 14 days after drug withdrawal,and the contents were significantly decreased in ZPT group,ZPT+ART group,2x ZPT+ART group and 3x ZPT+ART group(P < 0.05);Compared with ART group,serum levels of IL-6 and IL-10 in ZPT+ART group,2x ZPT+ART group,2.5 ZPT+ART group and 3x ZPT+ART group were decreased(P < 0.05),the content of TNF-α in T2.5 ZPT+ART group and 3 ZPT+ART group was decreased,and the content of TNF-α in 2 ZPT+ART group,2.5 ZPT+ART group and 3ZPT+ART group was significantly decreased(P < 0.05).4.On the 7th and 14 th day after drug withdrawal,compared with the control group,the average infection micee of mice in BEN group,ZPT group,2x ZPT+BEN group,2.5x ZPT+BEN group and 3x ZPT+BEN group were significantly decreased(P <0.05).The average infection micee of mice in ZPT+BEN group,2x ZPT+BEN group,2.5 ZPT+BEN group and 3x ZPT+BEN group was significantly decreased(P < 0.05);Compared with ZPT group,the average infection micee of mice in ZPT+BEN group,2x ZPT+BEN group,2.5 ZPT+BEN group and 3x ZPT+BEN group was significantly decreased(P < 0.05).The average inhibitory micee was significantly different among all groups(P < 0.05).Compared with blank group,liver and spleen coefficients increased in all groups,and there was no significant difference in the increase of liver and spleen coefficients in ZPT+ART group,2x ZPT+BEN group,2.5x ZPT+BEN group and 3x ZPT+BEN group compared with blank group(P > 0.05).There were significant differences between control group,BEN group and ZPT group compared with blank group(P < 0.05);Compared with the control group,the liver and spleen coefficients of the BEN group,ZPT group,ZPT+BEN group,2x ZPT+BEN group,2.5 ZPT+BEN group and 3x ZPT+BEN group were decreased,and the liver and spleen coefficients of the 3x ZPT+ART group were significantly decreased(P < 0.05);Compared with the control group,the serum contents of TNF-α,IL-6 and IL-10 in each group were decreased 14 days after drug withdrawal,and the contents were significantly decreased in ZPT group,ZPT+BEN group and 3x ZPT+BEN group(P <0.05);Compared with BEN group,serum levels of IL-6 and IL-10 in ZPT+BEN group,2x ZPT+BEN group and 2.5x ZPT+BEN group were decreased(P < 0.05),the TNF-α content in 2.5 ZPT+BEN group and 3 ZPT+BEN group was decreased,and the TNF-α content in 2 ZPT+BEN group,2.5 ZPT+BEN group and 3 ZPT+BEN group was significantly decreased(P < 0.05).Conclusion:In this experiment,we developed the 65 th genemiceion of artemisinin-resistant P.Berghei mouse malaria model with highly stable resistance.Based on this model,we found that: Dark Russell,the main active ingredient of dark Russell in Lingshuijiang,has obvious efficacy in the treatment of artemisinin-resistant P.berghei malaria and chloroquine-resistant P.berghei malaria,but there will be a certain degree of relapse.When dark Russell is combined with artesunate and benzol in the treatment of artemisinin-resistant P.berghei malaria and chloroquine-resistant P.berghei malaria,it is better than dark Russell alone.The therapeutic effect on mice is also increasing.Therefore,dark Russell has great development value as an antimalarial drug,and the combination of dark Russell and artemisinin drugs provides a new drug idea for the treatment of malaria. |
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