Effect Of Clostridium On The Expression Of PCNA,Ghrelin And Hsp70 In The Small Intestine Of Growing Pigs Induced By Deoxynivalenol

Deoxynivalenol(DON),also known as vomiting toxin(VT),poses a great threat to the healthy development of animal husbandry and causes huge economic losses to the livestock industry,however,it has high toxicity and stable molecular structure,and has strong acid resistance and heat resistance,therefore,there is no adsorbent product on the market which has significant detoxification effect on DON.The aim of this study was to validate the effect of a Clostridium as a detoxifying bacterium on the expression of PCNA,ghrelin and Hsp70 in DON-induced growth of porcine small intestine,and provide a detailed theoretical basis for the development and application of Clostridium.In this experiment,40 healthy PIC parents with a 4-month-old weight of 55.58 ±1.83 kg were randomly divided into 5 groups(n=8).The control group was fed a basal diet(DON:371.2 ?g/kg,AFB1:1.5 ?g/kg,FB1: 256.3 ?g/kg,ZEA: 145.16 ?g/kg),and Pure DON group fed basal diet + Vomiting toxin rice.(DON:5429.31?g/kg,AFB1: 3.42 ?g/kg,FB1: 283.98 ?g/kg,ZEA: 210.16 ?g/kg),Pure DON+Clostridium group: basal diet + Vomiting toxin rice +15mL/kgClostridium,natural DON group: basal diet + natural mildew wheat(DON: 1904.44 ?g / kg,AFB 1: 1.59?g/kg,FB1:325.7?g/kg,ZEA:196.08?g/kg),Natural DON+ Clostridium group: basal diet + natural moldy wheat + 15mL/kgClostridium.The pre-test period was 7 days and the test period was28 days.The duodenum,jejunum and ileum were slaughtered at the end of the experiment.HE and IHC and WB methods were used to observe the morphological changes of the small intestine and the changes of PCNA,ghrelin and Hsp70 protein distribution and expression,and analyze the detoxification effect of Clostridium on DON.The results:(1)Compared with the control group,the average daily gain of the pigs in the pure DON group decreased,and the average daily feed intake increased,but the effect was not significant(P>0.05).Histological observation showed that DON caused shortening of small intestine villi,increased depth of crypt,exfoliation of small intestinal mucosa,and duodenal injury was the most serious;IHC and WB results showed: DON inhibited the expression of PCNA in small intestinal mucosa and promoted the expression of PCNA protein in the small intestine gland,especially in the duodenum;DON improved the ability of small intestine villus mucosal epithelial cells and small intestine glands to secrete ghrelin and Hsp70.Ghrelin was most prominent in jejunum,and Hsp70 was most prominent in duodenum.The addition of Clostridium increased the average daily gain of pigs,the average daily feed intake tended tothe level of the control group,and the intestinal villus damage state was restored to a certain extent;the protein expression level of PCNA in the intestinal villus epithelium tends to the level of the control group,and the positive reaction of the small intestine gland is weakened,and the protein expression of ghrelin and Hsp70 is down-regulated,which is close to the control group.(2)The average daily gain and average daily feed intake of the natural DON group were significantly lower(P<0.05),natural DON significantly inhibits the growth and development of small intestine villi,resulting in different lengths of villi,shorter overall length,increased crypt depth,most obvious in duodenum.The results of IHC and WB showed that the DON toxin produced by natural mildew wheat inhibited the secretion of PCNA in the intestinal villus epithelium,while the positive reaction of the small intestine gland was enhanced,with the duodenum being the most prominent;The results of IHC and WB showed that the DON toxin produced by natural mildew wheat inhibited the expression of PCNA in the intestinal villus epithelium,while the positive reaction of the small intestine gland was enhanced,with the duodenum being the most prominent,and up-regulation of its protein expression,of which the ileum is most obvious;The addition of Clostridium significantly increased the average daily gain and average daily feed intake,which was close to the control level and played a role in restoring the morphological damage caused by natural DON;The results of IHC and WB showed that Clostridium increased the number of PCNA-positive cells in the intestinal villus epithelium,attenuated the positive reaction in the small intestine gland,and down-regulated the protein expression of ghrelin and Hsp70,which was close to the control group.Conclusion:Clostridium has a positive effect on the histological changes of the small intestine caused and the abnormal changes in the expression of PCNA,ghrelin and Hsp70 by DON.