Antagonistic Effect Of Sodium Selenite On Cadmium Chloride Induced Injury Of Porcine Alveolar Macrophages

The extensive damage mechanism of cadmium has toxic damage to many organs,systems and organs of the body.The sickle’s damage mechanism can lead to peroxidation in cells,mitochondria,DNA damage,and cytokine secretion disorder,which leads to oxidative stress in the body.Cadmium is a heavy metal,it is rich in nature,it is difficult to be degraded,can enter the human body along the food chain,and it is healthy for people with sores.Selenium has a variety of biological functions,including anti-inflammatory,enhancing immune function,relieving heavy metal toxicity and so on.In this study,the passage of porcine alveolar macrophages was cultured in vitro,and cadmium chloride（CdCl₂）and sodium selenite（Na₂SeO₃）were added alone or in combination to explore the anti-toxic effect of Na₂SeO₃ against CdCl₂ toxic damage.The research on the damage effect of cell phages and the protective effect of Na₂SeO₃ on CdCl₂ induced alveolar macrophage damage starts from two aspects.In order to provide new methods and ideas for future research on the damage effect of heavy metals and the development of therapeutic drugs,provide a theoretical basis for the biological function of selenium oxides against heavy metals,and provide a new idea for research in this field;at the same time,the healthy development of animal husbandry and the safety of human public health will be better.Test method:（1）CCK-8 method was used to detect different concentrations of CdCl₂（the initial concentration of CdCl₂ in the culture solution is:0,40,80,100,120,140,160,200,240μmol/L）and Different concentrations of Na₂SeO₃（the initial concentration of Na₂SeO₃ in the culture medium is:64,32,16,8,4,2,1,0.5μmol/L）for 24h to influence the viability of porcine alveolar macrophages,and calculate the24h When CdCl₂ induced the half-inhibitory concentration(IC₅₀)of porcine alveolar macrophages and the optimal concentration of Na₂SeO₃ on the cells;（2）According to the IC₅₀ of CdCl₂ and the optimal concentration of Na₂SeO₃,the test is divided into G1 group(IC₅₀),G2 group(1/2 IC₅₀),G3(1/4 IC₅₀),X group（8μmol/L Na₂SeO₃）,GX1 group,GX2 group,GX3 group;Detect the effect of CdCl₂ and Na₂SeO₃ on the viability of porcine alveolar macrophages by the CCK-8 method alone or in combination;（3）Add CdCl₂ and Na₂SeO₃ alone or in combination,culture for 24h in vitro,collect the cell supernatant,test the activity of LDH in the supernatant with the kit,and detect IL-1α,IL-4,IL-6 by ELISA method,IL-10,TNF-αcontent;（4）By adding CdCl₂ and Na₂SeO₃ alone or in combination,the cells and supernatant after24 hours of in vitro culture were collected,and the activity of LDH,SOD,CAT,GSH-PX and the content of MDA and GSH in the cells were detected by the kit.Results:（1）CdCl₂ and Na₂SeO₃ acted on porcine alveolar macrophages separately for 24h.The effects of both on cell viability were detected by the CCK-8method.It was concluded that the effects of CdCl₂ and Na₂SeO₃ on cells were both"dual".When cells are in a high-concentration cadmium（>80μmol/L）environment,CdCl₂ significantly inhibits cell viability（P<0.01）,while in a low-concentration cadmium（<40μmol/L）environment,CdCl₂ can promote cell viability;When the cells were exposed to low concentration of selenium（<16μmol/L）,it could promote the cell viability,but high concentration（>32μmol/L）showed a certain toxic effect.The IC₅₀ of CdCl₂ induced porcine alveolar macrophages was 106±0.02μmol/L;the optimal concentration of Na₂SeO₃ for porcine alveolar macrophages cultured in vitro was 8μmol/L,and the cell survival rate was 1.2197±0.1523%.（2）When CdCl₂alone acts on porcine alveolar macrophages for 24 hours,the cell survival rate decreases with the increase of CdCl₂concentration;compared with the control group,the G1 group（106μmol/L）significantly inhibited the cell activity（P<0.01）,G2 Group（53μmol/L）inhibited cell viability but not significantly（P>0.05）,and G3 group（26.5μmol/L）promoted cell viability.Na₂SeO₃（8μmol/L）alone acted on porcine alveolar macrophages for 24h.Compared with group B,Na₂SeO₃ significantly increased cell activity（P<0.01）.The combined addition of CdCl₂ and Na₂SeO₃cultured for 24h,compared with adding CdCl₂ alone,the cell viability was improved.（3）CdCl₂ alone acts on porcine alveolar macrophages for 24 hours.Compared the control group with the control group,cadmium can increase the activity of LDH in the cell culture supernatant,increase the content of IL-10 and IL-4,and decrease the content of IL-1αand IL-6.Decrease,the content of TNF-αincreases in high-concentration cadmium environments,and the content decreases in low-concentration cadmium environments;Na₂SeO₃ alone acts on porcine alveolar macrophages for 24 hours.Na₂SeO₃ alone porcine alveolar macrophage 24h,compared with the control group,the supernatant IL-10,IL-4,IL-6,TNF-αcontent significantly improved（P<0.05）,the content of IL-1αbut not significantly increased（P>0.05）;the same time,the supernatant IL-1αwas significantly decreased（P<0.05）,LDH activity decreased（P>0.05）.After adding sodium selenite combined,the content of the cell culture supernatant IL-10,IL-4 levels were reduced,IL-1αand IL-6levels rise.It is effective against TNF-αin porcine alveolar macrophages caused by high concentrations of Cd.The increase in content has a certain antagonistic effect;it can also have an antagonistic effect on the decrease of TNF-αcontent in porcine alveolar macrophages caused by low concentration of Cd.（4）CdCl₂ alone acts on porcine alveolar macrophages for 24 hours,resulting in a decrease in the activities of SOD,CAT,and GSH-PX in the cells,which all decrease with increasing concentrations;the contents of MDA and GSH in the cells increase.When Na₂SeO₃acts on porcine alveolar macrophages alone,it increases the activity of SOD,CAT,GSH-PX and the content of GSH in the cells;reduces the content of MDA in the cells.After the combined addition of sodium selenite,the activities of SOD,CAT,and GSH-PX in cells increased,the content of MDA decreased,and the content of GSH decreased.Conclusion:CdCl₂ can damage alveolar macrophages and decrease cell viability.The IC50 of CdCl₂ and Na₂SeO₃ on porcine alveolar macrophages was 106±0.02μmol/L;the optimal concentration of Na₂SeO₃ was 8μmol/L;8μmol/L Na₂SeO₃could protect porcine alveolar macrophages from injury induced by CdCl₂（106μmol/L,53μmol/L,26.5μmol/L）effect.Generally speaking,selenium has anti-inflammatory and anti-oxidation effects,and has an antagonistic effect on the injury of porcine alveolar macrophages induced by CdCl₂.