Effects of Diquat on Oviduct Magnum Structure,Cell Apoptosis and Expression of Genes Related to Reproductive Performance in Hens(Gallus gallus)
Diquat,a bipyridine herbicide commonly utilized in modern agricultural production,can easily induce oxidative stress in poultry,in order to investigated the effects of oxidative stress induced by Diquat on hens'oviduct magnum,in this study,45 healthy 180-day-old Kangle yellow hens(Gallus gallus)with similar body weight were randomly divided into 3 groups,with 15 hens in each group.The hens in each group were fed a basal diet and had free access to water.After 1 week of adaptive feeding,the experimental groups were injected with 10 and 20 mg/kg BW(body weight)dose of Diquat intraperitoneally,while those in control group were injected with an equal dose of normal saline intraperitoneally.After 21 d,the oviduct magnum tissues were collected.The antioxidant indicators were detected using Antioxidant Function Assay Kit.The pathological injury was observed using hematoxylin-eosin(HE)staining technology,and the relevant protein distribution were detected by immunohistochemistry(IHC)technology.The expressions of apoptosis and reproductive performance-related genes and proteins were detected using qPCR and Western blot technology.The results showed that the activities of catalase(CAT)and total superoxide dismutase(T-SOD)in the oviduct magnum of Diquat groups were significantly decreased(P＜0.05),and the concentration of malondialdehyde(MDA)was significantly increased(P＜0.05).HE staining results showed that the tube diameter of oviduct magnum in Diquat groups were reduced,cells of lamina propria were sparsely-distributed,cilia in the free surface of epithelium mucosae were damaged,and mucosal epithelial cells were vacuolated in Diquat 20 mg/kg group.IHC results showed that the staining intensity of proliferating cell nuclear antigen(PCNA)in the epithelium mucosae and lamina propria of oviduct magnum in Diquat groups were decreased.The staining intensity of Wingless-related murine mammary tumor virus integration site 4(Wnt4)in the free surface of mucosal epithelium was also decreased.qPCR results showed that the gene expression level of anti-apoptosis-related gene B-cell lymphoma-2(Bcl-2)in oviduct magnum tissues of Diquat 20 mg/kg group was significantly decreased(P＜0.05).The gene expression levels of Bcl-2 associated X protein(Bax)and cysteinyl aspartate specific proteinases 3(Caspase3)and the ratio of Bax/Bcl-2 in Diquat groups were significantly up-regulated(P＜0.05).The gene expression levels of reproductive performance-related genes zona pellucida sperm-binding protein 2(ZP2)and Wnt4 were significantly down-regulated(P＜0.05).Western blot results showed that the protein expression levels of Bcl-2,Bax,Caspase3,ZP2 and Wnt4 were basically consistent with the gene expression changes.The above results indicated that Diquat can induce oxidative stress and apoptosis in the oviduct magnum of hens,thereby damaging the tissue structure of the enlarged part and inhibiting the expression of genes related to reproductive performance of the oviduct magnum.This study provides a theoretical basis for alleviating the damage to the oviduct of poultry caused by the bipyridine pesticides and protecting the reproductive performance.
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