ABSTRACT
Objective To investigate the effect of transient receptor potential vanilloid 4 (TRPV4) inhibitor HC067047 on anxiety-like behavior in mice induced by lipopolysaccharide (LPS). Methods Totally 48 male C57BL/6 mice were randomly divided into control group (NS), model group (LPS) and drug intervention group (HC + LPS). Anxiety mouse model was established by intraperitoneal injection of 0.83 mg/kg LPS. The HC + LPS group was intraperitoneally injected with HC067047 (10 mg/kg) 30 minutes before LPS injection, and the NS group and LPS group were injected with equal volume of normal saline. Open field test and social interaction experiments were used to detect anxiety-like behaviors in each group of mice; Immunohistochemical chemistry and Western blotting were used to detect the expression of TRPV4, inducible nitric oxide synthase (iNOS), neuronal nitric oxide synthase (nNOS) and endothelial nitric oxide synthase (eNOS) in the hippocampus. Results Immunohistochemical and Western blotting experiments showed that, compared with the NS group, the expression of TRPV4 in the hippocampus of the LPS group was significantly up-regulated (P<0.0001); In the open field test, compared with the NS group, the total distance (P < 0.0001), the distance in the central area (P<0.01) and the time of in the central area mice in the LPS group reduced significantly (P< 0.01). HC067047 intervention reversed the activities of LPS model mice total distance (P < 0.05), the distance of activities in the central area (P < 0.001) and the time of in the central area (P < 0.01); In the social interaction test, compared with the NS group, the interaction time the unfamiliar mice reduced significantly in LPS group (P<0.01), which was reversed by HC067047 treatment (P< 0.01); Western blotting detection revealed that the expression of hippocampal iNOS (P<0.05), nNOS (P < 0.001), and eNOS (P < 0.001) in the LPS group were significantly higher than the NS group, which reduced remarkably by HC067047 treatment (iNOS P < 0.05, nNOS P < 0.01 and eNOS P < 0.01). Conclusion Inhibiting the expression of TRPV4 can improve the anxiety-like behavior, and this process may be related to anti-oxidative stress.
ABSTRACT
Aim To investigate the effect of high-salt model of mouse thoracic aortic endothelial cells on the production of nitric oxide (NO) . Methods After preincubation of thoracic aortic endothelial cells with transient receptor potential vanilloid 4 (TRPV4) inhibitor HC067047, the effects of TRPV4 on NO production were studied by Ca2+and NO staining with calcium ion fluorescent probe Fluo-4 and NO fluorescent probe DAF-FM DA. The thoracic aortic endothelial cells were stimulated with a high salt concentration of 60 mmo! L"1 to detect Ca2+influx and NO production Compared with control group, suppression of TRPV4 inhibited Ca2+influx and NO production in thoracic aortic endothelial cells, and high salt conditions inhibited TRPV4-medicated Ca2+influx and NO production compared with mannitol under the same osmotic pressure. Conclusion In high salt state, the inhibition of TRPV4 channel leads to the decrease of Ca2+influx and the down-regulation of NO synthesis.