The present research introduces the method of Production of M2000 (β-d-
mannuronic acid) and its
therapeutic effect on experimental model of
nephritis. M2000 was produced using enzymatic and chemical procedure on prepared
alginate from Pseudomonas fluorescens. The experimental
glomerulonephritis was induced in rats by a subcutaneous immunization and daily
intravenous administration of
bovine serum albumin (BSA). M2000
solution (30mg/kg) was administered intraperitoneally at regular 48-h intervals for 4 weeks. Onset of treatment was day 56. Urinary
protein was measured weekly and serum anti-BSA antibody was assessed by ELISA method at different intervals. Animals were killed on day 84 and blood samples and kidney specimens were obtained. Serum (
creatinine, BUN,
cholesterol, and
triglyceride) and urine (
protein,
urea, and
creatinine) determinants were measured at the time of sacrifice. Kidney specimens were processed for light and immunofluorescent microscopic examination. The
fibrosarcoma cell line was used for assaying tolerability and
matrix metalloproteinase type 2 (MMP-2) activity. MMP-2 activity was assessed using zymography. Our data showed that M2000
therapy could significantly reduce the urinary
protein excretion in treated rats versus non-treated controls. Anti-BSA antibody titer was lower in treated rats than in controls at the 12th experimental week. PMN infiltration and glomerular
immune complex deposition was less intense in treated rats than in controls. Cytotoxicity analysis of M2000 showed a much higher tolerability compared with other tested drugs (
diclofenac,
piroxicam and
dexamethasone). The inhibitory effect of M2000 in MMP-2 activity was significantly greater than that of dexsamethasone and of
piroxicam at a concentration of 200μg/ml. Moreover, the toxicological study revealed that M2000 had no influence on serum (BUN,
creatinine,
triglyceride and
cholesterol) determinants, urinary
protein excretion and glomerular histology in healthy group receiving
drug.
CONCLUSIONS: In this research, for the first time we introduced the procedure of production of M2000 (β-d-
mannuronic acid) and our data suggest that treatment with M2000, as a novel anti-inflammatory
drug can reduce
proteinuria, diminish antibody production and suppress the progression of disease in experimental model of
glomerulonephritis.