Bacillus Amyloliquefaciens B10 Can Alleviate Aflatoxin B1-induced Kidney Oxidative Stress And Apoptosis In Mice

As a natural toxin contained in food,aflatoxin is a great threat to animals.It can damage the liver,kidney and other tissues of the animal body,and can cause cancer in the animal in severe cases.Aflatoxin B1(AFB1)is the most dangerous and the most widely studied among aflatoxins,and is classified as a carcinogen by the International Agency for Research on Cancer.Since there is no specific antidote for AFB1 at present,it is a research hotspot to explore ways to reduce the toxicity of AFB1 to the body.Due to the probiotic properties of probiotics,more and more people are used to study the detoxification effect of mycotoxins.Therefore,this study intends to use the Bacillus amyloliquefaciens B10 isolated and identified in our laboratory as the research object to explore whether it can alleviate the kidney injury induced by AFB1 in mice and its mechanism.Methods: In this experiment,56 6-week-old male Kunming mice were randomly divided into 4 groups,each with 14 mice.They were respectively Con group,AFB1 group,AFB1+B10 group and B10 group.The mice were fed by gavage.Gavage with LB culture fluid in group Con;Gavage AFB1 working fluid in AFB1 group,Gavage B10 working fluid + AFB1 working fluid in AFB1+B10 group,Gavage B10 working fluid in group B10,in which AFB1 and B10 both use LB culture fluid as a carrier.Gavage the mice.After 28 days of intragastric administration,the mice were euthanized,blood was collected from the eyeballs,and serum was collected by centrifugation to detect the kidney function indexes UA,BUN and CRE of the mice.Obtain fresh kidney tissue,rinse with saline,and remove excess fat on it.Weighing is used to determine the ratio coefficient of mouse organs.Select some fresh kidney tissues,fix them with 4% paraformaldehyde,and perform HE staining and Tunel detection.The remaining kidney tissue was chopped up and stored in a refrigerator at-80°C.For MDA,GSH-PX,SOD and CAT oxidative stress index detection and Western Blot detection(Nrf2,HO-1,PTEN,AKT,P-AKT,Keap-1,Bcl-2,Bax,Caspase-3 and Caspase-9).Result:1.The results of the mouse organ ratio coefficient showed that compared with the Con group,the mouse organ ratio coefficient in the AFB1 group increased significantly(P<0.01);compared with the AFB1 group,the kidneys of the mice in the AFB1+B10 group The converter ratio coefficient is reduced(P<0.01).The results show that B10 can effectively alleviate the kidney damage caused by AFB1.2.The serological test results of mice showed that compared with the Con group,the content of UA,BUN and CRE serological indicators in the AFB1 group was extremely significantly increased(P<0.01);compared with the AFB1 group,the AFB1+B10 group The serum levels of UA,BUN and CRE in mice decreased(P<0.01).The results show that B10 can effectively alleviate the serum changes in mouse kidney induced by AFB1.3.The test results of the oxidation kit showed that compared with the Con group,the MDA content of the AFB1 group was extremely significantly increased(P<0.01),and the vitality of GSH-PX,SOD and CAT was extremely significantly reduced(P<0.01);compared with AFB1 Compared with the group,the content of MDA in the AFB1+B10 group was reduced(P<0.01),and the activities of GSH-PX,SOD and CAT were significantly increased(P<0.01).The results show that B10 can effectively inhibit the oxidative stress damage of the kidney induced by AFB1.4.Pathological tissue section results showed that renal interstitial congestion appeared in the kidney tissue of the AFB1 group of mice;vacuole-like lesions appeared in the renal tubules;renal tubular epithelial cells swelled and died,and the sloughed epithelial cells formed cell casts;renal cysts The cavity space was wider than that in the Con group;severe granular lesions and extensive hemorrhage appeared in the kidney tissue.After B10 treatment,the kidney bleeding of mice was reduced.The results show that B10 can improve the kidney damage caused by AFB1.5.Tunel test results showed that compared with the Con group,the number of apoptotic cells in the kidney tissue of the AFB1 group was extremely significantly increased(P<0.01);compared with the AFB1 group,the number of apoptotic cells in the kidney tissue of the AFB1+B10 group was extremely significant Decrease(P<0.01).The results show that B10 can effectively inhibit AFB1 induced apoptosis.6.Western Blot detection of related proteins showed that the results of the Nrf2/HO-1 signaling pathway showed that compared with the Control group,the expression of Nrf2 protein in the AFB1 group was extremely significantly reduced(P<0.01);the expression of HO-1 protein was significantly reduced(P<0.05);Keap-1 protein expression increased significantly(P<0.01).The results of apoptosis signal pathway showed that compared with the Control group,the protein expression of AKT,P-AKT and Bcl-2 in the AFB1 group was significantly reduced(P<0.05);PTEN,Bax,Caspase-9 and Caspase-3 The protein expression level of the protein increased significantly(P<0.01).After B10 treatment,this result was reversed(P<0.01;P<0.05).The results showed that B10 can regulate Nrf2/HO-1 and apoptosis signaling pathway to protect mouse kidney tissue from AFB1-induced oxidative damage and apoptosis.Conclusion: Bacillus amyloliquefaciens B10 has an antagonistic effect on the kidney damage of male mice induced by AFB1.Bacillus amyloliquefaciens B10 can effectively reduce AFB1-induced kidney tissue damage in mice through Nrf2/HO-1 and apoptosis pathways.