| Pesticide residue is one of the most important research in food safety. It has become a new research direction to improve ability response to oxidative stress by using natural antioxidants in food. The present study hypothesis soy isoflavone(SIF) could protect against commonly used carbamate pesticide carbaryl-induced toxicity in nerve cells. PC12 cells were used as a model for nervous cell system. PC12 cells were exposed to carbaryl, treated with genistein(GEN), daidzein(DAI), or their mixed solution for indicated times. It has been investigated that the protective effect of SIF could against carbaryl-induced toxicity in PC12 cells from the decrease of cytotoxicity, neurotransmitter normalization, and change in oxidative stress level.(1) Carbaryl can induce neurotoxicity in PC12 cells. In this study, PC12 cells were exposed to carbaryl in different concentrations from 0.00 ?g/m L to 200.00 ?g/m L, and showed significant changes. Carbaryl-induced cell injuries produced excessive amounts of ROS. At the same time, carbaryl can interact with acetylcholinesterase(ACh E), and inhibit ACh interact with ACh E. ACh released from PC12 cells to medium, and also produced excessive amounts of ROS. Relative fluorescence intensity(RFI) increased from 37.38±11.48 to 45.56±11.96 firstly, then decreased to 17.11±1.50 with the damage of cells. Excessive ROS increased mitochondrial membrane potential(MMP) depolarization ratio, increased the activity of Caspase-3 and the leakage rate of LDH. At the same time the cellular antioxidant system were changed, malondialdehyde(MDA) content gradually increased from 19.27±0.96 ?mol/mg·prot to 112.39±1.59 ?mol/mg·prot, superoxide dismutase(SOD) activity increased from 4.08±0.87 U/mg·prot to 17.77±0.43 U/mg·prot, and the inhibition rate increased from(19.00±1.66)% to(73.00±1.71)%, glutathione(GSH) content reduced from 96.15±6.10 ?mol/g·prot to 22.91±3.98 ?mol/g·prot. Then lead to cell morphological changes, cell became shrinkage, synaptic smaller, and villi disappeared. Finally with the increase of carbaryl concentration, the PC12 cells survival rate decreased from(100±0.46)% to(27.86±1.69)%.(2) Genistein can increase ACh metabolism in PC12 cells. 20.00 ?g/m L GEN has no cytotoxicity in PC12 cells, however, it has pro-oxidant effects on PC12 cells with the content of MDA increased to 13.19±6.69 nmol/mg prot, and RFI level reached 112.51±8.21. At the same time, GSH content increased to 195.89±0.00 ?mol/g·prot. When PC12 cells were exposed to carbaryl, treated with GEN could accelerate the metabolism of ACh, reduce the excessive ACh caused the damage by improving alpha 7-n ACh Rs activity. What's more, GEN can act as a inhibitor of PTK, which inhibit the oxidation of ROS by inhibiting the PTK related pathway.(3) Daidzein could improve PC12 cell antioxidant enzyme system activity. 20.00 ?g/m L DAI has no cytotoxicity, and improved PC12 cell antioxidant enzyme system activity. GSH increased to 297.27±9.47 ?mol/g·prot. DAI reduction is less than GEN in chemical experiment due to the lack of one hydroxyl on C-5 position. However, DAI has better lipotropy in vitro. On the surface of cell membrane, DAI concentration is higher than GEN due to low block conformation. When PC12 cells exposed to carbaryl, treated with DAI reduce the content of MDA and SOD inhibition rate, RFI value, induced the increase of GSH content, SOD activity effect through its antioxidant properties. DAI could also inhibit Caspase-3 mediated apoptosis pathway to achieve the purpose of protecting the nerve.(4) SIF can effectively mitigate adverse effects induced by carbaryl. GEN and DAI have a combined effect that is greater than the effect obtained with each isoflavone alone. Bacause of the low concentration of GEN, mixed solution has better antioxidant activity. PC12 cells GSH content increased to 269.90±0.00 ?mol/g·prot, and LDH leakage rate deceased. When PC12 cells exposed to carbaryl, treated with GEN-DAI not only inhibiting apoptosis mediated by Caspase-3, but improving the activity of alpha 7-n ACh Rs allosteric, enhance nerve cell signaling, and then antagonized the inhibition of TK excess ACh produced by ROS, and finally induce PC12 cells enhanced the antioxidant activity. SIF enhances metabolism of choline and plays a role as both antioxidant and neuroprotectant. In this study, we suggest that consuming a variety of traditional soy foods seems more effective at preventing some diseases than supplementing a normal diet with a pure soy isoflavone mixture.In this research, protective effects of soy isoflavnone genistein, and daidzein against carbaryl-induced toxicity in PC12 cells were researched. Based on this, it could provide direct experimental evidence and possible interventions for cell antioxidant protection pesticide injury, has important theoretical significance. |
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