Over-expression of arginine vasopressin in magnocellular neurosecretory cells of hypothalamus and its potential relationship with development of diabetic nephropathy

Figures

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  Figure 1. Rat body weight in diabetes mellitus (DM) and control groups at 0, 4, 8, 12, 16, 20 and 24 weeks. N = 18 in DM group and n = 10 in control group Data are shown as mean ± SD, *p < 0.05 and **p < 0.01 compared to control group.

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  Figure 2. Morphological studies of diabetic kidney in rats. A – Light microscopic findings of renal morphological changes in control rats by periodic acid-Schiff staining (PAS 400×). B – Light microscopic findings of renal morphological changes in diabetes mellitus (DM) rats by PAS staining (400×). C – Electron microscopic findings of renal ultramicrostructure changes in control rats (12 000×, scale bar: 2 μm). D – Electron microscopic findings of renal ultramicrostructure changes in DM rats (12 000×, scale bar: 2 μm). N = 18 in DM group and n = 10 in control group

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  Figure 3. Morphological studies of hypothalamus in rats. A, B – Morphological studies of hypothalamus in control and diabetes mellitus (DM) rats. C – Number of neurons in hypothalamus in DM group and control group. D – Gray value of astrocytes in hypothalamus in DM group and control group. E – Number of GFAP positive cells in hypothalamus in DM group and control group. N = 18 in DM group and n = 10 in control group Data are shown as mean ± SD, *p < 0.05 and **p < 0.01 compared to control group.

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  Figure 4. Expression levels of AVP in hypothalamus and levels of RVP in kidney of rats. A – The qRT-PCR analysis of expression of AVP mRNA in hypothalamus of diabetes mellitus (DM) rats and controls. B – The qRT-PCR analysis of expression of RVP mRNA in kidney of DM rats and control rats. GAPDH served as the loading control of qRT-PCR. C – Immunofluorescence analysis of AVP expression in hypothalamus of DM rats and controls. AVP was stained in red (Cy3-conjugated), and nuclei were stained with Hoechst-33258 dye (blue). Average optical density (AOD) value of AVP in hypothalamus of DM rats is shown below. Scale bar: 20 μm. D – Immunochemistry analysis of RVP expression in kidney of DM rats and normal controls. AOD value of RVP in kidney of DM rats is shown below. Scale bar: 20 μm. E – Western blot analysis of AVP protein expression level in DM rats and control rats. F – Western blot analysis of RVP protein expression level in DM rats and control rats. β-actin served as the loading control of Western blot. N = 18 in DM group and n = 10 in control group Data are shown as mean ± SD, **p < 0.01 compared to control group.

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  Figure 5. The correlation between AVP, RVP expression and proteinuria. A – The positive correlation between proteinuria and mRNA expression of AVP. B – The positive correlation between proteinuria and protein expression of AVP. C – The positive correlation between proteinuria and mRNA expression of RVP. D – The positive correlation between proteinuria and protein expression of RVP. E – The positive correlation between mRNA expression of AVP and RVP. F – The positive correlation between protein expression of AVP and RVP. N = 18 in DM group and n = 10 in control group