Effects Of Primary Chick Kidney Cell Culture And Expression Of Related Genes Caused By NIBV Infection

To investigate the changes of morphological, antioxidant function and apoptosis genes mRNA expression of chick kidney(CK) cells underlying the nephropathogenic infectious bronchitis virus(NIBV) infection, We presented a primary culture model based on the mechanical and enzymatic dissociation of sterile kidney tissue obtained from the SPF chickens(14 days old). The CK cells were divided into 2 groups(infected and control) and inoculated with 0.2 mL sterile DMEM and virus at 72 h post-incubation, respectively. At 12,24, 36, 48, 60, 72 hours post-infected(hpi), the cell viabilities were tested by MTT; the cells and mediums were harvested to investigate the changes of LDH, XOD, UA, MDA and SOD levels in cell and medium by commercial kits; the cells were harvested to determine the relative mRNA expression levels of XOD, Bak1, Bcl2, p53, Caspase-3 and DDIT3 and absolute quantification of N gene of NIBV, individually. The results show that there was a time-dependent increase in lactate dehydrogenase(LDH) both in cells and medium following NIBV infection. Meanwhile, NIBV infection of CK cells resulted in dysregulation of antioxidant function of CK cells, as superoxide dismutase(SOD) activity was decreased and malonaldehyde(MDA) concentration was increased in cells while there was elevation of SOD activity in medium. Furthermore, the xanthine oxidase(XOD)activity and the uric acid(UA) concentration of infected group were significantly increased both in cell and medium, when cell XOD mRNA transcription showed increased in infected group compared with the control group. Taken together, The generation of chicken kidney cells proliferation was inhibited underlying the NIBV infection, the metabolic disorder of XOD and anti-oxidation function were belong to the important pathological mechanism of NIBV infection, the high mRNA expression of apoptosis genes indicate NIBV infection caused apoptosis of primary CK cells.