| Aflatoxins have always been a threat to livestock production because of their wide distribution and toxicity.And for cows,aflatoxin can indirectly threaten human health through milk.Resveratrol has been extensively researched in the past two decades and has shown a positive effect on animal.In this study,MAC-T cells were used as research objects,and a series of studies on the MAC-T cells were carried out on the effects of AFB1 and resveratrol.The aim is to understand whether resveratrol can counteract the effects of AFB1 on MAC-T cells and how the optimal conditions.MAC-T cells were treated with different concentrations of AFB1 and/or resveratrol.Then the effects of the cells were evaluated for their cell viability,morphology and expression of resistancegenes.And the appropriate treatment concentration was screened for the next study.In this study,the treatment of MAC-T cells with different concentrations of AFB1 resulted in a significant decrease in cell viability at 24 h of AFB1 greater than or equal to 4 mg/L;likewise at 4 mg/L AFB1 treatment MAC-T cells showed a significant decrease in total antioxidant capacity and mitochondrial membrane potential;however,AFB1 did not appear to cell morphology and cytoskeleton.Quantitative expression of inflammatory factors and partial transcription of Nrf2 signaling pathway by qPCR showed that significant concentration-correlated change in TNF-α,IL-1β and IL-6,but IL-8 transcription was significantly up-regulated when treated with greater than or equal to 6 mg/L AFB1.However,the transcription of Nrf2 and Keap1 genes did not increase significantly at 24 h.On the contrary,the transcription level of downstream anti-stress gene NQO1 increased with the concentration of AFB1;at 10 mg/L,HSP70 was significantly down-regulated.In the same way,the corresponding items were tested of the treatment by resveratrol,and it was found that the cytotoxicity increased with the increase of resveratrol concentration,and decreased significantly when it reached 30 mg/L.When the resveratrol treatment concentration was 40 mg/L,the total antioxidant capacity of the cells increased,and the mitochondrial membrane potential was hyperpolarized at all treatment concentrations.It is worth noting that resveratrol showed inhibition of cell proliferation at any concentration treatment.At the same time,different resveratrol treatment concentrations have a two-way effect on the MAC-T cytoskeleton.That is,when the resveratrol treatment concentration is less than 10 mg/L,the edge of the cell is promoted to form a flaky pseudopod,but at a treatment concentration higher than 20 mg/L,the flaky pseudopod at the cell edge is inhibited.Similar to AFB1,there was no consistent effect on the transcription of inflammatory factors.TNF-α was significantly down-regulated;IL-1β increased significantly with increasing treatment concentrations;IL-8 was up-regulated at treatment concentrations above 40 mg/L.However,IL-6 was down-regulated at a high treatment concentration and up-regulated at low treatment concentrations.Significant increases in gene transcription in the Nrf2 signaling pathway were observed with greater than 20 mg/L of resveratrol.Based on the above results,4 mg/L AFB1 and 10 mg/L resveratrol were selected for cross-test.The results showed that resveratrol can alleviate the cytotoxicity induced by AFB1,including reducing the increased ROS of MAC-T cells under the action of AFB1,restoring mitochondrial membrane potential and reducing apoptosis.Expression of mRNA transcripts in the Nrf2 signaling pathway was determined by qPCR,and resveratrol also showed good regulation.Therefore,it was shown that resveratrol has a resistant effect on aflatoxin toxicity in MAC-T cells.Conclusion: Resveratrol can resist the damage of AFB1 in MAC-T cells.However,it is necessary to control the dose of resveratrol used because high doses of resveratrol are still toxic. |
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