nephrin and Hypercholesterolemia

The protective effects of Rho kinase inhibitor fasudil against renal diseases have recently been reported. We compared the therapeutic effects of fasudil on the spontaneously hypercholesterolemic (SHC) rat, a model of chronic kidney disease (CKD) with proteinuria, with those of the angiotensin receptor blocker olmesartan (OL) by paying attention to the proteinuria and the macrophage phenotype. SHC rats were allocated to six treatment groups: a vehicle (Ve) group, a low-dose fasudil (FL) group, a high-dose fasudil (FH) group, an OL group, a combination of low-dose fasudil and OL (CL) group, and a combination of high-dose fasudil and OL (CH) group. Sprague-Dawley rats treated with vehicle served as a control (n = 7/each). The rats were treated for 24 wk. Compared with the Ve group, proteinuria was significantly decreased in the FH, OL, and CL groups, and it completely disappeared in the CH group. Glomerular stainings of nephrin and F-actin were focally impaired in the Ve group but were restored in the CH group. Western blotting showed that the CH group had significantly increased renal nephrin expression compared with the Ve group. Interstitial infiltration of macrophages was significantly increased in the Ve group, which was significantly attenuated in all treatment groups. The ratio of CD206 (M2 macrophage marker) to CD68 mRNA was significantly greater in the CH group than in the Ve group. These results indicate that fasudil with OL reduces proteinuria by protecting podocyte integrity and alters the interstitial macrophage density/phenotype, thereby exerting renoprotective effects against CKD.

Topics: 1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; Actins; Angiotensin II Type 1 Receptor Blockers; Animals; Antigens, CD; Antigens, Differentiation, Myelomonocytic; Enzyme Inhibitors; Hypercholesterolemia; Imidazoles; Kidney Glomerulus; Lectins, C-Type; Macrophages; Male; Mannose Receptor; Mannose-Binding Lectins; Membrane Proteins; Microscopy, Electron; Phenotype; Proteinuria; Rats; Receptors, Cell Surface; Renal Insufficiency, Chronic; rho-Associated Kinases; RNA, Messenger; Tetrazoles

The mouse, as opposed to the rat, is relatively resistant to the experimental nephrosis induced by puromycin aminonucleoside. The reason for this species specificity is not known. Apolipoprotein E (apoE)-deficient mice were used to determine whether hypercholesterolemia plays a role in inducing proteinuria.. Thirty-two mice were divided into normal and high cholesterol diet groups and then divided further into four subgroups: puromycin, puromycin+probucol, probucol and control. Urinary albumin of these mice was analyzed by nephelometry. The lipid peroxidation (LPO) end products malonyldialdehyde (MDA) and 4-hydroxynonenal (4-HNE) were detected by immunohistochemistry, and the expression level of the glomerular slit diaphragm protein, nephrin, was studied by immunohistochemistry and real time RT-PCR.. Overt proteinuria was induced by puromycin only in the apoE knockout mice ingesting the high cholesterol diet. The staining intensities of MDA and 4-HNE were stronger in the glomeruli of proteinuric mice compared to glomeruli of non-proteinuric mice. When serum cholesterol levels were reduced by probucol, proteinuria decreased and fewer LPO end products were seen immunohistochemically. Three and eight days after puromycin injection the level of nephrin mRNA in the kidneys of proteinuric mice decreased in comparison to the controls. Puromycin-treated mice kidneys demonstrated a clearly reduced reactivity to the nephrin antibodies.. Hypercholesterolemia, possibly via LPO, is a prerequisite for puromycin-inducible glomerular damage in the mouse. Furthermore, nephrin protein and mRNA levels appear to be candidate markers of glomerular damage in the mouse.

Topics: Animals; Apolipoproteins E; Cholesterol, Dietary; Hypercholesterolemia; Immunohistochemistry; Lipid Peroxidation; Membrane Proteins; Mice; Mice, Knockout; Nephrosis; Protein Synthesis Inhibitors; Proteins; Proteinuria; Puromycin; RNA, Messenger