Establishment Of Paraquat Poisoning Model And Study On Detoxification Effect Of Coenzyme Q10

Paraquat is widely used in agricultural production in China.It is a highly toxic quaternary ammonium herbicide,which is highly toxic to humans and animals.Paraquat poisoning has become a serious public health problem to be solved urgently in society because of having no special antidote,still mainly requiring supportive treatment and having a poor prognosis,which result in an extremely high fatality rate after poisoning.In this study,based on the two model organisms of Caenorhabditis elegans(C.elegans)and mice,the animal model of paraquat poisoning was established to explore the feasibility of coenzyme Q10(CoQ10)as an antioxidant in the treatment of paraquat poisoning.1.Constructing a model of C.elegans poisoned by paraquat(1)Gradient concentration of paraquat acted on C.elegans.Through the statistical analysis of C.elegans survival,swallowing ability and movement ability,Paraquat concentration of 5 mg/mL was selected to construct the model of paraquat poisoning.Paraquat concentration of 5 mg/mL significantly shortened the life span of C.elegans.(2)CoQ10 gradient concentration acted on paraquat poisoning C.elegans model.The results showed that CoQ10 prolonged the lifespan of paraquat poisoning C.elegans in a dose-dependent manner,and showed good treatment effect on paraquat poisoning C.elegans.(3)Possible mechanism of CoQ10 on paraquat poisoning C.elegans: The DCFH-DA fluorescent probe was used to detect the ROS content in the C.elegans.CoQ10 pretreatment can reduce ROS content in paraquat poisoning C.elegans and inhibit ROS rising levels;The mRNA expression levels of eight kinds of oxidative stress genes(gcs-1),glutathione S-transferase(gst-4,gst-20),superoxide dismutase(sod-1,sod-2,sod-3,sod-4,sod-5)in C.elegans were detected by real-time fluorescence quantitative(qPCR).CoQ10 pretreatment can inhibit the increase of mRNA expression of some genes of glutathione S-transferase gene family and superoxide dismutase gene family in paraquat-induced C.elegans,and inhibit oxidative stress in paraquat-poisoned C.elegans;By fluorescence imaging of transgenic C.elegans CL2166(GST-4::GFP).CoQ10 pretreatment reduced the level of oxidative stress in paraquat-poisoned C.elegans;The content of MDA in C.elegans was detected by TBA colorimetry.CoQ10 pretreatment could reduce the level of lipid peroxidation and reduce the degree of oxidative damage in paraquat-poisoned C.elegans.2.Construction of a mouse model of paraquat poisoning(1)Different doses of paraquat were intragastrically administered to mice.According to the survival conditions of mice,paraquat was selected to construct a mouse model of paraquat poisoning.The dose of paraquat was 80 mg/kg.(2)CoQ10 was administered orally to poisoned mice,and the mortality of mice was examined to study the effect of CoQ10 on paraquat poisoned mice.CoQ10 administration can significantly improve symptoms such as shortness of breath,sluggish action,and malaise in paraquat poisoning mice,while increasing the chance of survival.CoQ10 shows good treatment effect on paraquat poisoning mice(3)CoQ10 gavage administration of paraquat poisoning mice.By calculating the wet/dry weight ratio of lung tissue in mice with paraquat poisoning,the administration of CoQ10 could reduce the wet/dry weight ratio of lung tissue in mice with paraquat poisoning,and to some extent alleviate the pulmonary edema in mice with paraquat poisoning;The content of ROS in the lung tissue of mice was detected by DCFH-DA fluorescence probe.The administration of CoQ10 could reduce the content of ROS in the lung tissue of mice poisoned by paraquat to some extent;The SOD activity in lung tissue of mice was detected by WST-1 colorimetry.CoQ10 administration could improve SOD activity in lung tissue of paraquat poisoning mice;The MDA content in lung tissues of mice was detected by TBA colorimetry.CoQ10 can reduce the MDA content in lung tissues of paraquat poisoned mice to a certain extent.(4)In order to further detect the effect of CoQ10 on lung injury in mice poisoned by paraquat,paraffin sections of lung tissue were made and pathological analysis was carried out.It was found that CoQ10 administration could significantly relief the pulmonary tissue structure disorder of paraquat poisoning mice,the intra alveolar bleeding and the infiltration of inflammatory cells,and reduce lung injury in paraquat poisoning mice.In summary,the results show that CoQ10 plays an important antioxidant role,which can effectively remove excess ROS produced in the body,reduce oxidative stress,and have a better anti-lipid peroxidation effect,which can reduce paraquat-induced oxidative damage and reduce the degree of damage caused by paraquat poisoning to the body,CoQ10 shows a good treatment effect on the two model organisms after paraquat poisoning,and it has a certain protective effect on the lungs,which are the main organs damaged by paraquat poisoning.