Effect of Native and Modified Lipoproteins, and Albumin on Human Mesangial Cell Proliferation / 대한신장학회잡지

BACKGROUND: Modified lipoproteins may be involved in nephro- and glomerulosclerosis. Diabetic nephropathy-like lesions have also been induced in a rat model by glycated and glycoxidized albumin. In cultured rat or human mesangial cells, enhanced cell proliferation and production of mesangial matrix in response to lipoproteins and their modified forms have been demonstrated by [3H]-thymidine incorporation and cell counting assays. But these methods are relatively complex and most of them have used only one or two of the lipoprotein, albumin and their modified forms. METHODS: We investigated the effects of native and modifed lipoproteins, and albumin on cultured human mesangial cell proliferation using non-radioactive colorimetric method by MTS/PMS assay. Lipoproteins added were low density lipoprotein(LDL), high density lipoprotein(HDL), very low density lipoprotein(VLDL), oxidized LDL(oxidation with copper sulfate in vitro) and glycated LDL and we also used albumin, glycated albumin, and interleukin-1beta as a positive control. RESULTS: Interleukin-1beta promoted the proliferation of cultured human mesangial cells up to concentration 20 ng/mL. LDL induced the proliferation of mesangial cells in a concentration-dependent manner up to concentration 100 microgram/mL. HDL and VLDL had no significant proliferative effect. Oxidized LDL caused the proliferation of mesangial cells at low concentration up to concentration 25 microgram/mL. Addition of glycated LDL resulted in a concentration- dependent inhibition of mesangial cells. Albumin and glycated albumin inhibited the proliferation of mesangial cells at low concentration of 100 microgram/mL, but cell growth was increased at higher concentrations. CONCLUSION: We demonstrated the effects of the single and modified proteins on the proliferation of cultured human mesangial cell by relatively simple colorimetric method. Results were almostly identical to those of previous studies obtained by radioactive method or cell counting assay.

Screening of antimicrobial peptide from marine clamworm Marphysa sanguinea and preliminary research on its antitumor effect / 中国海洋药物

Active fractions were obtained from the c(?)amworm homogenate by reverse-phase concentration, gel filtration and affinity column chromatography. The MTS/PMS assay was used in the screening of antimicrobial peptide in vitro. The MTS/PMS assay and ELISAAFP method were combined to evaluate the effects of antimicrobial peptide on the proliferation and ?-fetoprotein secretion of HepG2 cells as well as on the proliferation of normal mouse osteoblast MC3T3-E1. The results showed that the purified antimicrobial peptide from clamworm Murphysa sanguinea was a constitutive basic protein with MW 8.1 kDa and pI 8.6, which showed inhibition effects on the proliferation and ?-fetoprotein secretion of HepG2 cells at different levels. These effects have a positive correlation with the concentrations of antimicrobial peptide. No obvious inhibition and damage effects were observed on the normal osteoblast.