Study on Asiatic Acid Alleviates LPS-induced Acute Kidney Injury by Regulating Apoptosis and Autophagy of Broilers
This study aimed to investigate the effect of Asiatic acid(AA)on lipopolysaccharide(LPS)-induced apoptosis and autophagy in the kidneys of broilers.Forty broilers(one day old)were fed adaptively until seven days old and then were randomly divided into the control group(Con),LPS group(LPS),low dose AA group(LPS+AA 15 mg·kg-1)and high dose AA group(LPS+AA 30 mg·kg-1).The broilers in the AA pretreatment group were gavaged with corresponding doses of AA daily for 14 consecutive days.Except for the control group,broilers in other groups were intraperitoneally injected with 0.5 mg·kg-1 LPS at the age of 16,18 and 20 days to establish an acute kidney injury(AKI)model.All broilers were sacrificed at the age of 21 days,and kidney samples were collected.Hematoxylin-eosin(HE)staining was used to observe the histopathological changes in the kidney tissue.The kidney coefficient was calculated and the levels of antioxidant enzymes in kidney tissue were detected.Real-time fluorescence quantitative(qRT-PCR)and Western blot were used to detect the expression of apoptosis-related genes and proteins as well as the expression of autophagy-related genes and proteins.Immunohistochemis-try and immunofluorescence were used to detect the expression and distribution of cytochrome c(Cytc)and microtubule-associated protein 1A/1B-light chain 3(LC3)in kidney tissue.The TUNEL staining was used to detect the apoptosis rate of the kidneys.The results showed that AA could alleviate LPS-induced kidney pathological damage of broilers,and significantly reduce the kidney coefficient and malondialdehyde(MDA)content as well as significantly increase the activities of glutathione peroxidase(GSH-Px),superoxide enzyme(SOD)and catalase(CAT)(P＜0.05).AA significantly reduced the mRNA expression of P53,BAX and Caspase3 in LPS-induced AKI of broilers(P＜0.05),and significantly promoted the mRNA expression of BCl2,Beclin1,ATG5,LC3-Ⅰ and LC3-Ⅱ in LPS-induced AKI of broilers(P＜0.05).Moreover,AA pretreatment significantly reduced the protein expression of P53,Bak1,BAX and Cleaved-Caspase3 in LPS-induced AKI of broilers(P＜0.05),and significantly promoted the protein expression of Beclin1,ATG5,LC3 Ⅱ/Ⅰ in LPS-induced AKI of broilers(P＜0.05).Immunohis-tochemical and immunofluorescence results also showed that AA pretreatment significantly reduced the protein expression and distribution of Cytc in LPS-induced AKI(P＜0.001),and significantly promoted the protein expression and distribution of LC3 in LPS-induced AKI(P＜0.001).The TUNEL results showed that AA pretreatment significantly reduced the LPS-induced apoptosis rate in the kidney of broilers(P＜0.001).In summary,AA alleviates LPS-induced AKI of broilers by inhibiting oxidative stress,promoting renal cell autophagy and reduc-ing apoptosis.This result provides a theoretical basis for AA becoming a potential feed additive and preventing LPS-induced AKI of broilers.
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维普
