To investigate whether acetoacetic acid(ACAC)induced mitochondrial damage of bovine mammary epithelial cells by inhibiting the nuclear transcription factor NF-E2-related factor 2(Nrf2).Bovine mammary epithelial cells(MAC-T)were treated with 0,0.6,1.2 or 1.8 mmol·L-1 ACAC for 24 h;MAC-T cells were pretreated with 10 μmol·L-1 SFN for 24 h and then treated with 1.2 mmol·L-1 ACAC for 24 h.Flow cytometry was used to detect mitochondrial reactive oxygen species(ROS);JC-1 staining was used to detect mitochondrial membrane potential changes;the gene expression of mitochondrial oxidative phosphorylation complexⅠ-Ⅴ(COⅠ-Ⅴ)was determined by q-PCR,as well as the mitochondrial biosynthesis regulator Nrf2,peroxisome proliferator-activated receptor γ-γ activator 1-α(PGC-1α),mitochondrial transcription factor A(TFAM),Nuclear respirator-1(Nrf1),and mitochondrial DNA;the adenine nucleoside triphosphate(ATP)content was detected by the kit.The results showed that after ACAC treatment,the content of ROS was significantly increased(P<0.05),while the mitochondrial membrane potential decreased significantly(P<0.05),and the mRNA expression level of COⅠ,COⅡ,COⅢ,COⅣ and COⅤ were significantly decreased(P<0.05),the mRNA expression of PGC-1α,TFAM,Nrf1 and Nrf2 were decreased(P<0.05),and the mtDNA content was decreased(P<0.05),the ATP content was decreased(P<0.05),while SFN pretreatment alleviated the above effects.The results showed that ACAC induced mitochondrial damage in MAC-T cells by inhibiting the Nrf2 signaling pathway.
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