The Protective Effect And Mechanism Of Quercetin On Oxidative Damage And Cell Apoptosis Under ?-ZOL Exposure

?-Zearalenol(?-Zearalenol,?-ZOL)is the most important metabolite of the mycotoxin zearalenone(ZEN)in humans,animals and plants.It is mainly found in corn,wheat,rice and oats and other agricultural products have multiple organ toxicity,and its toxicity is about 10 times that of the parent toxin ZEN.The influence and control of its toxicity is one of the current research hotspots.One of the research trends is to use the biological activity of food-derived natural ingredients to prevent and control the toxicity of hazardous factors.Quercetin(Quercetin,Que)is a common dietary polyphenol flavonoid compound widely found in vegetables and fruits.It has anti-oxidation,anti-apoptosis and anti-inflammatory activities,and has important protective effects on the human body.In order to study the preventive and protective effect of Que and the improvement effect of Que on the body damage after?-ZOL exposure,this article uses two application simulation scenarios of?-ZOL after Que preventive treatment and Que intervention after?-ZOL exposure to experimental animals.Caenorhabditis elegans and Caco-2 cells were studied separately,and the protective effects of Que on cell oxidative damage and apoptosis under?-ZOL exposure under two simulation scenarios were further studied,and the protective mechanism was carried out.explore.The main research results and conclusions are as follows:(1)This paper studies for the first time in two application simulation scenarios of?-ZOL infection after Que preventive treatment and Que intervention after?-ZOL exposure.Que's effect on the growth status of model animals Caenorhabditis elegans and Caco-2 cells after?-ZOL exposure(Using indicators such as nematode body length,egg production,and cell proliferation activity)protection and intervention.The results show that in the?-ZOL exposure simulation scenario(Scenario 1)after Que preventive treatment,the IC₅₀ of?-ZOL is 82.94?mol/L.At the non-damaging concentration of Que(80?mol/L),the low concentration of Que(20?mol/L,40?mol/L)pretreatment has a protective effect on the growth and reproduction of nematodes exposed to different doses(20,40,60?mol/L)of?-ZOL.Among them,40?mol/L Que pretreatment can increase the relative body length of nematodes treated with 20,40,and60?mol/L?-ZOL by 2%,7%and 8%,respectively,and increase the relative egg production of nematodes by 21%,75%,25%;when Que exceeds 60?mol/L,it has no protective effect on the growth of nematodes.40?mol/L Que pretreatment can significantly increase the survival rate of Caco-2 cells treated with 20,40,and 60?mol/L?-ZOL by 19%,23%,and 19%,and return to normal levels,but when the concentration of ZOL is higher than 60?mol/L,Que pretreatment has no effect on cell survival;In the Que intervention simulation scenario(scenario 2)after?-ZOL exposure,low-dose(20,40?mol/L)Que intervention has an improvement effect on nematodes injured by low-dose?-ZOL.40?mol/L Que can increase the relative body length of nematodes infected with?-ZOL(20 and 40?mol/L)by 2%and 10%,respectively.The egg volume is increased by 2%and 13%,but when the concentration of?-ZOL is too high(60?mol/L),the growth and reproduction ability of nematodes cannot be improved.At the same time,40?mol/L Que can significantly increase the survival rate of cells treated with 20,40,and 60?mol/L?-ZOL by 13%,4%,6%,but when the concentration of?-ZOL reaches 60?mol/L,Que intervention also has no effect on the damaged cells.Therefore,Que pretreatment and intervention within a certain concentration range can alleviate the damage caused by?-ZOL to cells and nematodes to a certain extent.Among them,40?mol/L Que pretreatment has a more significant protective effect on nematodes and cells than Que intervention.(2)The experiment explored the protective effect and mechanism of Que pretreatment on?-ZOL-induced cell damage through the study of cell oxidative damage,apoptosis and related protein expression in the toxin exposure simulation scene after Que pretreatment.In terms of cell oxidative damage,low-dose(20,40?mol/L)Que pretreatment can significantly eliminate excess ROS generated in cells caused by low-dose(20,40?mol/L)?-ZOL damage,and its relative fluorescence intensity It is reduced by 11?39%;the relative activity of antioxidant enzyme SOD is increased by5?23%;the relative content of lipid peroxidation product MDA is reduced by 12?24%.Compared with the 40?mol/L?-ZOL injury group without Que pretreatment,40?mol/L Que pretreatment significantly increased the number of cells,and the relative protein content in the cells increased by 12%.In terms of apoptosis and protein expression,40?mol/L Que pretreatment can increase the relative level of ATP and mitochondrial membrane potential in cells by 3%and 9%,respectively,and reduce the apoptosis rate by 3%,reducing the fragmentation and fragmentation of nuclei.Intensive staining reduces the degree of DNA damage and inhibits the occurrence of cell apoptosis;at the same time,Que pretreatment can rationally regulate the anti-apoptotic protein Bcl-2 and the pro-apoptotic protein Caspase-3,and maintain the normal physiological activities of cells.It shows that Que pretreatment can exert its antioxidant effect,slow down cell oxidative stress,and protect the integrity of mitochondrial membrane,thereby protecting cells from oxidative damage and apoptosis caused by?-ZOL.(3)Experiments through the study of cell oxidative damage,apoptosis and related protein expression in the simulated scene of Que intervention after?-ZOL toxin exposure,further explore the improvement effect of Que intervention on?-ZOL-induced cell damage and its mechanism.Low-dose(20,40?mol/L)?-ZOL treatment can cause overproduction of ROS in cells,increase of MDA content,and decrease of antioxidant enzyme SOD.Re-intervention with low concentrations(20,40?mol/L)of Que after poisoning can obviously reverse the effect of?-ZOL damage on cells,reducing the relative fluorescence intensity of ROS in cells by 30?61%and the relative content of MDA 10?33%,the relative activity of SOD increased by 11?18%.40?mol/L?-ZOL treatment can change the morphology of cells and cell nuclei,reduce cell number and protein content,reduce ATP levels and mitochondrial membrane potential and a series of cell damages,until apoptosis.In addition,40?mol/L Que intervention can reduce the apoptosis caused by?-ZOL,reduce the appearance of irregular fragmented cells,and increase the relative protein content,ATP relative level and mitochondrial membrane potential in cells by 13%,13%,and 19%respectively,the cell apoptosis rate is reduced by 2%.Que intervention can stimulate the nuclear transfer of Nrf2,promote the production of antioxidant enzymes,and at the same time down-regulate the expression of pro-apoptotic protein Caspase-3,inhibit chromosome aggregation,and thus play an anti-apoptotic effect.This shows that Que intervention protects cells from the toxic damage of?-ZOL by improving the integrity of the mitochondrial membrane,regulating cell structure and cutting DNA repair between nucleosomes.In summary,both Que pretreatment and intervention can alleviate the toxic effects of cells and nematodes exposed to?-ZOL to a certain extent.Que pretreatment can enhance ROS clearance by regulating the Bcl-2/Casepase-3 apoptotic pathway.Ability and self-stress resistance,reduce oxidative damage,and protect cells to a certain extent;Que intervention promotes the production of antioxidant enzymes by stimulating the nuclear transfer of the antioxidant pathway Nrf2,and at the same time down-regulates the expression of the pro-apoptotic protein Caspase-3 Cell damage.