Asiatic Acid Alleviates LPS-induced Pyroptosis in Renal Cell by Inhibiting HMGB1/TLR4/NF-κB Pathway in Broilers
The aim of this study was to explore the mechanism of asiatic acid(AA)on LPS-in-duced pyroptosis in kidney cells of yellow-feather broilers through HMGB1/TLR4/NF-κB path-way.Forty one-day-old healthy broilers were reared to 7 days of age and randomly divided into four groups:control group(CON),LPS-induced model group(LPS),AA low-dose group(LPS+AA 15 mg·kg-1)and AA high-dose group(LPS+AA 30 mg·kg-1).Broilers in the AA treat-ment groups were given the corresponding dose of AA by intragastric administration for 14 days.Except the control group,all broilers were injected intraperitoneally 0.5 mg·kg-1 LPS to estab-lish the model of acute kidney injury at 16,18 and 20 days of age.Broilers were killed on the 20th day after LPS was intraperitoneally injected for 12 h and renal samples were collected.He-matoxylin-eosin(HE)staining was used to observe the pathological changes of renal tissue.The mRNA expression levels of HMGB1,TLR4,NF-κB,IκB,NLRP3,Caspase-1,IL-18,IL-1βand TNF-α in renal tissue were detected by RT-qPCR.Protein expression levels of HMGB1,TLR4,P-NF-κB,NLRP3,GSDMD,IL-1β and Cleaved caspase-1/Caspase-1 were detected by Western blot.The expression level of P-IκB was detected by immunohistochemical method.The protein expression levels of HMGB1,NLRP3 and ASC were detected by immunofluorescence.The results showed that LPS induced vesicular degeneration of renal tubular epithelial cells and glomerular dysplasia,which could be ameliorated by treatment with AA.At the same time,the mRNA levels of HMGB1,TLR4,NF-κB,IκB,NLRP3,Caspase-1,TNF-α and IL-18 in re-nal tissue were significantly downregulated by AA(P＜0.05).Protein levels of HMGB1,TLR4,P-NF-κB,NLRP3,GSDMD,IL-1β,and Cleaved caspase-1/Caspase-1 decreased.The results of immunohistochemistry and immunofluorescence showed that AA could significantly reduce the expression and distribution of P-IκB,HMGB1,NLRP3 and ASC in renal tissue by LPS-induced.In conclusion,AA can alleviate LPS-induced pyroptosis in kidney cells of broilers by inhibiting the HMGB1/TLR4/NF-κB pathway.
万方数据
维普
