ABSTRACT
OBJECTIVE: To elucidate the regulation of p38 and PKC for water channel protein AQP9 and the effect on As intake in lour kinds of mammal cells, and to investigate the regulation mechanism of AOP9 phosphorylation. METHODS: p38, PKC protein and phosphorylation levels were analyzed by Western blotting, and phosphorylation level of AQP9 was detected by immuno-precipitation. Intracellular arsenic content was determined by inductively coupled plasma mass spectrometry (ICP-MS). Experimental data were analysed by SPSS statistical software. RESULTS: p38 protein and phosphorylation levels increased significantly with time in four kinds of cells treated with NaAsO2, while no significant change had been observed in PKC protein and phosphorylation levels. AQP9 phosphorylation level was inhibited in L-02 cells when p38 activity was inhibited, and As accumulation also decreased significantly in L-02 cells. CONCLUSION: AQP9 phosphorylation has effect on As intake and rate. But the regulated mechanism may be different in different cells. p38 kinase attends partly AQP9 phosphorylation in liver normal cells L-02, PKC has no effect on AQP9 phosphorylation in all kinds cells used in this paper.
ABSTRACT
Objective:To explore the expression and role of AQP9 in the models of cultured steatosis hepatocytes.Methods:Steatosis models of hepatocytes was established by adding oleic acid to the growing L-02 cell strain.The expression of AQP9 mRNA was measured with RT–PCR and the protein expression of AQP9 were measured by Western blotting.Lipid droplets in the hepatocytes were observed by oil red staining and the contents of triglyceride,FFA and glycerol in hepatocytes were measured with analyzed kit.Results:Through oil red staining.a few lipid droplets were observed at 24h and steatosis hepatocyte increased greatly at 72h.Triglyceride,FFA and glycerol contents in hepatocytes of model groups also increased,as compared with the control group(P
ABSTRACT
Objective:To study the role of AQP9 and its mRNA expression in the formation of ischemic cerebral edema in the rats.Methods:MCAO models were established by occluding unilateral middle cerebral artery(MCA) of the rats with the suture method.The morphological changes were observed with HE stain and transmission electron microscope(TEM).Brain water content(BWC) was measured by using wet-dry weighing method.The expression of AQP9 and its mRNA in the edema brain tissue were detected with in situ hybrilization(ISH) and immunohistochemistry(IHC).Results:The space around the neurons and perivascular space became larger.Some neurons showed necrosis phenotype,and the number of the neurogliocyte increased.The expression of AQP9 and its mRNA were up-regulated in subfornical organ(SFO)、choroid plexes、brain cortex、supraoptic nuclei、hippocampus and other regions,and reached its peak at 48~72h after MCAO.The brain water content(BWC) in ischemic cerebral tissue reached its peak at 72h.Conclusions:the expression of AQP9 and its mRNA were up-regulated following the brain edema.The results imply that AQP9 might play an important role in the formation of ischemic cerebral edema.