PMID: 11788780Jan 15, 2002Paper

Induction of metallothionein-I protects glomeruli from superoxide-mediated increase in albumin permeability

Experimental Biology and Medicine
Ram SharmaV J Savin


Metallothioneins (MT) are low-molecular-weight, heat-stable, cysteine-rich proteins with four isoforms. MT-I and MT-II are ubiquitous and are induced by oxidative, physical, and chemical stress. MT-I is an efficient scavenger of superoxide (*O2) and hydroxyl ion (OH(-)). We have demonstrated that *O2 and hypohalous acid can cause an increase in glomerular albumin permeability (P(alb)) in vitro. The purpose of this study was to document the protective effect of MT gene product on the *O2-mediated increase in P(alb). Glomeruli from Sprague-Dawley rats in 4% BSA medium were incubated for 4 hr at 37 degrees C in duplicate tubes. Each set contained glomeruli alone or with 5 microM Cd(++), 0.3 mM Spermine-NONOate (NO donor), 0.3 mM Sulfo-NONOate (nitrous oxide donor), 0.6 mM SNP (nonspecific NO donor) and SNP + carboxy-PTIO (10 mg/ml). After incubation, one set of tubes was used to isolate total RNA for the measurement of the mRNA levels of MT-I by reverse transcriptase polymerase chain reaction (RT-PCR). Duplicate tubes were incubated for an additional 10 min with 10 nM of *O2, and P(alb) was measured using video microscopy. RT-PCR of total RNA from Cd(++) and Spermine-NONOate treated glomeruli revealed a 2-fold induction of MT-I ex...Continue Reading


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