坏死梭杆菌对奶牛真皮成纤维细胞线粒体动力学与自噬的影响

doi:10.16431/j.cnki.1671-7236.2026.01.037

Abstract

Abstract:

Objective This study was conducted to investigate the effects and potential mechanisms of Fusobacterium necrophorum subsp. necrophorum （Fnn） infection on mitochondrial dynamics and mitophagy of bovine dermal fibroblasts（BDFs）. Method This study isolated and cultured BDFs from the skin between cow toes and co-cultured them with 1×10⁸ CFU/mL Fnnfor 12 h to establish an infection model group （Fnn）. Uninfected cells were used as control group （CON）， the morphological changes of BDFs afterFnninfection were observed. The cell activity of BDFs was detected by CCK-8 kit. Real-time quantitative PCR was used to detect the changes in the expression of genes related to mitochondrial dynamics， biosynthesis and mitophagy， and immunofluorescence assay was used to detect the changes in the expression of related proteins. Result Compared with CON group， cells in Fnn group showed swelling， bulging， and deformation， and Fnn infection significantly inhibited the activity of BDFs. Real-time quantitative PCR results showed that compared with CON group， after Fnn infection， the relative expression levels of peroxisome proliferator receptor gamma coactivator alpha （PGC-1α）， nuclear respiratory factor-1 （Nrf1）， mitochondrial transcription factor A （Tfam）， mitochondrial fusion factor gene-2 （Mfn-2）， phosphatase and tensin homolog induced protein kinase 1 （PINK1）， E3 ubiquitin ligase （Parkin） genes in BDFs were significantly or extremely significantly decreased （P<0.05 or P<0.01）， while the relative expression levels of optic nerve atrophy protein 1 （Opa1）， Mfn-1， and Sequetosome 1 （P62） genes showed a decreasing trend （P>0.05）. The relative expression of genes Fis1， light chain-3α （LC3α）and LC3β were extremely significantly increased （P<0.01）. The results of immunofluorescence assay detection showed that compared with CON group， after Fnn infection， the fluorescence signals of dynein related protein 1 （Drp1）， PINK1， Parkin and P62 in BDFs were enhanced， the fluorescence signals of Opa1， Mfn-1 and LC3 were weakened. Conclusion Fnn infection could disrupt the mitochondrial biosynthesis of BDFs， disrupt mitochondrial fusion and fission processes， and induce mitophagy. This results provided a new theoretical basis for a deeper understanding of the pathogenesis of hoof rot disease.

Key words: foot rot; Fusobacterium necrophorum subsp.necrophorum; mitochondrial biosynthesis; mitochondrial dynamics; mitophagy

CLC Number:

S852.61

LIU Meng, ZHANG Anchi, DING Ruxin, GE Yansong, ZHENG Jiasan. Effects of Fusobacterium necrophorum subsp. necrophorum on Mitochondrial Dynamics and Mitophagy of Bovine Dermal Fibroblasts[J]. China Animal Husbandry & Veterinary Medicine, 2026, 53(1): 418-426.

Figures/Tables 6

Table 1

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引物名称 Primer name	引物序列 Primer sequences （5′→3′）	登录号 Accession No.	退火温度 Annealing temperature/℃	产物长度 Product length/bp
PINK1	F：CTGCAGTTGCCTTTCACCAG	XM_061148080.1	57.6	185
PINK1	R：CCCTGGCCGTAAAAGGGATT	XM_061148080.1	57.6	185
Parkin	F：CTGGGACCAGAGCGGAATTT	XM_070795610.1	57.7	110
Parkin	R：GCAGCTGATGTCTCGACTGT	XM_070795610.1	57.7	110
P62	F：TCAGCTTCTGCTTTAGCCCG	XM_061149642.1	57.7	165
P62	R：AAAAGGCAACCAAGTCCCCA	XM_061149642.1	57.7	165
LC3α	F：CCACTCAGGCCTTCTTCCTG	XM_027558755.1	58.2	95
LC3α	R：ATCCTCGTCCTTCTCCTGCT	XM_027558755.1	58.2	95
LC3β	F：ATGCCGTCCGAGAAAACCTT	XM_061386957.1	57.3	112
LC3β	R：TTGCCATACTTCCTTCCGGG	XM_061386957.1	57.3	112
PGC-1α	F：GGACATGTGCAACCAGGACT	XM_045474382.1	57.8	65
PGC-1α	R：GGTCTTCACCAACCAGAGCA	XM_045474382.1	57.8	65
Nrf1	F：ATGACCATCCAGACAACGCA	XM_027539015.1	57.1	151
Nrf1	R：CCTCTTTTATTGCCCACCCCT	XM_027539015.1	57.1	151
Tfam	F：TGAATGCGCTGGGAAAGTCA	NM_001034016.2	57.1	71
Tfam	R：ACTAAAGGGATAGCGCAGTCG	NM_001034016.2	57.1	71
Opa1	F：GCCTGACATTGTGTGGGAGA	XM_045502573.1	57.7	159
Opa1	R：CCAGGTGAACCTGTGGTGAA	XM_045502573.1	57.7	159
Mfn-1	F：GCTGGGCTTCTCTTGTCCAT	XM_061415356.1	57.7	64
Mfn-1	R：AATCCTAGAAGCACCCGCTG	XM_061415356.1	57.7	64
Mfn-2	F：GGGCTCAGAGGAGAAGAGGA	XM_061160381.1	58.2	72
Mfn-2	R：AGCTGTTCGTCCTGGTGAAG	XM_061160381.1	58.2	72
Fis1	F：GCTGAACGAGTTGGTGTCTG	XM_059038717.2	56.7	157
Fis1	R：AGCAAGGCCTTTACGGATGT	XM_059038717.2	56.7	157
β-actin	F：TCCTGCGGCATTCACGAAACTAC	XM_027528015.1	60.7	80
β-actin	R：GTGTTGGCGTAGAGGTCCTTGC	XM_027528015.1	60.7	80

Effects of Fusobacterium necrophorum subsp. necrophorum on Mitochondrial Dynamics and Mitophagy of Bovine Dermal Fibroblasts

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