Arbidol reduces neurotoxicity induced by aquaporin-4 antibody positive serum in vivo and in vitro

Abstract

Objective To investigate the protective effect of arbidol on aquaporin-4(AQP4)antibody-induced neurotoxicity in vivo and in vitro. Methods (1)The cortical cells were conventionally obtained from newborn SD rats; 4 days after seeding, the cultured cortical cells were randomly divided into four groups: control group, AQP4 antibody-positive serum treatment group, arbidol preconditioning group and dimethyl sulfoxide(DMSO)treatment group; cells in the arbidol preconditioning group were subdivided into 12.5 μmol/L, 25 μmol/L and 50 μmol/L arbidol preconditioning subgroups; cells in the control group were given 10% volume ratio of healthy human sera; cells in the AQP4 antibody-positive serum treatment group were added the same volume of AQP4 antibody-positive serum; cells in the 12.5 μmol/L, 25 μmol/L and 50 μmol/L arbidol preconditioning subgroups were given arbidol one h before adding AQP4 antibody-positive serum. After 6 h of cultivation, the morphological and total area changes of astrocytes and neurons through Hoechst33258/PI staining and immunofluorescence were aboserved.(2)Twenty-four SD rats were randomly divided into control group, AQP4 antibody-positive serum treatment group, arbidol preconditioning group and DMSO treatment group; rats in the later three groups were induced animal models of neuromyelitis optica(NMO)via intracerebral injection of AQP4 antibody positive serum; rats in the later two groups were given 54 mg/kg(concentration: 10 mg/mL)arbidol and DMSO respectively, 3 h before adding AQP4 antibody-positive serum; NMO models were identified by HE and immunohistochemical staining. Three d after convention feeding, the rats in the four groups were sacrificed, and the brain tissue sections were performed immunohistochemical staining to detect the expression levels and lesion size of AQP4 and glial fibrillary acidic protein(GFAP). Results (1)In vitro, the cell viability in 25 μmol/L arbidol preconditioning subgroup was significantly increased as compared with AQP4 antibodypositive serum treatment group and other subgroups(P<0.05). The total areas of positive cells(astrocytes and neurons)in AQP4 antibody-positive serum treatment group were significantly decreased as compared with those in the control group(P<0.05); while those in the arbidol preconditioning group was significantly increased as compared with those in the AQP4 antibody-positive serum treatment group(P<0.05).(2)In vivo, intracerebral injection of AQP4-Ab positive serum caused NMO-like pathology, with leukocyte infiltration and loss of AQP4 and GFAP within the lesion; the loss areas of AQP4 and GFAP in AQP4 antibody-positive serum treatment group were significantly increased as compared with those in the control group(P<0.05); while those in the arbidol preconditioning group was significantly decreased as compared with those in the AQP4 antibody-positive serum treatment group(P<0.05). Conclusion Arbidol alleviates the neurotoxicity which induced by AQP4-Ab positive serum in vivo and in vitro, and it would be effective on the treatment of NMO. Key words: Neuromyelitis optica; Aquaporin-4; Arbidol; Astrocyte; Neuron; In vivo; In vitro