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This study aimed to investigate the effects of Escherichia coli （E. coli）， Clostridium butyricum （C. butyricum）， and their metabolites on mitochondrial function and related metabolic pathways in porcine intestinal epithelial cells （IPEC-J2）， providing a theoretical basis for regulating intestinal health. Initially， bacterial suspensions were screened to ensure logarithmic-phase growth （3 h） with optimal viability for subsequent experiments. IPEC-J2 cells were then treated with varying concentrations of Escherichia coli， Clostridium butyricum， lipopolysaccharide （LPS）， and short-chain fatty acids （SCFAs）， with 3 replicates per group. Cell viability and morphological integrity were assessed to determine the optimal treatment duration and concentration. Key metrics included ATP levels， reactive oxygen species （ROS）， mitochondrial membrane potential， and the expression of genes related to antioxidant defense and mitochondrial function. The results showed that： 1） The optimal concentrations for treating IPEC-J2 cells were determined as 108 CFU·mL-1 for Escherichia coli and 106 CFU·mL-1 for C. butyricum， with a treatment duration of 1 h. The treatment duration of SCFAs and LPS were also treated for 1 h. 2） Propionic acid and E. coli significantly increased ROS levels （P<0.05）， whereas C. butyricum and LPS reduced them （P<0.05）. 3） C. butyricum upregulated the expression of ACC1， NRF1， TFAM， HO-1， APAF-1， PIG3， and p53 （P<0.05）； E. coli downregulated the expression of TFAM and CtyC （P<0.05） but upregulated the expression of p53 （P<0.05）； Propionic acid and acetic acid had no significant effects on genes expression. The results indicated that SCFAs enhanced cellular metabolism， E. coli and LPS triggered oxidative stress and apoptotic pathways. C. butyricum activated mitochondrial lipid metabolism， antioxidant defenses， and apoptosis in IPEC-J2 cells. These findings offer novel insights into mitochondria-targeted strategies for gut microbiota regulation.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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