Emodin ameliorates glucose-induced matrix synthesis in human peritoneal mesothelial cells

Abstract

Our findings demonstrate that emodin ameliorates the undesirable effects of concentrated glucose on HPMC via suppression of PKC activation and CREB phosphorylation, and suggest that emodin may have a therapeutic potential in the prevention or treatment of glucose-induced structural and functional abnormalities in the peritoneal membrane.

Anti‐DNA antibody induction of protein kinase C phosphorylation and fibronectin synthesis in human and murine lupus and the effect of mycophenolic acid

“…Except where indicated otherwise, cells were washed 3 times with PBS between all steps, and all incubations were at 37°C for 1 hour. Cells were blocked with 3% BSA in PBS and incubated with primary antibody to FN in a humidified chamber (23). Cells were subsequently incubated with the relevant FITC-conjugated secondary antibody (1:100 dilution), mounted, and viewed under epifluorescence using an Axioskop 2 plus fluorescence microscope.…”

“…Whole cell lysate preparations were obtained by solubilizing human mesangial cells cultured under control or experimental conditions in 20 mM sodium acetate, pH 6.0, containing 4M urea and 1% Triton X-100 (200 l). The lysates (10 g of total protein content) were subjected to electrophoresis under denaturing conditions on 8% acrylamide gels to investigate FN synthesis and on 12% acrylamide gels to investigate PKC and MAPK activation (23). Samples were transferred onto nitrocellulose membranes and immunoblotted with primary antibodies to FN, ␤-actin, PKC␣, PKC␤I, PKC␤II, ERK, JNK, and p38 MAPK as previously described (23).…”

“…Bioactive TGF␤1 was assayed using a mink lung epithelial cell (MLEC) bioassay as previously described (23). A standard curve was constructed from the varying degrees of MLEC growth inhibition observed with standard concentrations of recombinant human TGF␤1 (23).…”

“…The lysates (10 g of total protein content) were subjected to electrophoresis under denaturing conditions on 8% acrylamide gels to investigate FN synthesis and on 12% acrylamide gels to investigate PKC and MAPK activation (23). Samples were transferred onto nitrocellulose membranes and immunoblotted with primary antibodies to FN, ␤-actin, PKC␣, PKC␤I, PKC␤II, ERK, JNK, and p38 MAPK as previously described (23). The bands were visualized by enhanced chemiluminescence, semiquantified by densitometry using ChemiGenius analysis software (Syngene, Cambridge, UK), and expressed as arbitrary densitometric units (DU).…”

“…A standard curve was constructed from the varying degrees of MLEC growth inhibition observed with standard concentrations of recombinant human TGF␤1 (23).…”