Intracellular lactate dehydrogenase concentration as an index of cytotoxicity in rat hepatocyte primary culture

Chao, E; Dunbar, Deborah; Kaminsky, Laurence S.

doi:10.1007/bf00141283

Cited by 36 publications

(13 citation statements)

References 19 publications

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“…Intracellular LDH activity was assayed by measuring NADH consumption during the conversion of pyruvate to lactate (Chao et al 1988). The reduction ofNAD § was monitored at 340 nm with a Hitachi spectrophotometer in the presence of lactate.…”

Section: Methodsmentioning

confidence: 99%

Neuron and glial cell marker proteins as indicators of heavy metal-induced neurotoxicity in neuroblastoma and glioma cell lines

et al. 1993

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The sensitive and specific biochemical indicators for assessing chemical-induced neurotoxic insults in cell culture models have not been sufficiently explored. This study was designed to assess the usefulness of glia-specific beta-S100 protein and neuron-specific enolase (NSE) as indices of in vitro neurotoxicity of heavy metals. Glioma C6 and neuroblastoma N18TG-2 cells were grown in Dulbecco's modified Eagle's medium containing various concentrations of mercuric chloride (HgCl2) or cadmium chloride (CdCl2) for 5 days. Toxic response patterns of the neurospecific endpoints (beta-S100 and NSE), which were monitored with enzyme immunoassays, were compared with those of the non-neurospecific endpoints such as cell viability, total cellular protein, lactate dehydrogenase (LDH) activity, and cumulative glucose consumption in the two cell lines. Both HgCl2 and CdCl2 produced dose-dependent inhibition of neurospecific endpoints and non-specific endpoints. However, by ranking the EC50 values (effective concentration producing half-maximal inhibition) for various endpoints, the lowest values were found for beta-S100 in C6 cells, and for NSE in N18TG-2 cells. In lower and intermediate concentrations, the inhibitory effects of the heavy metals on the content of beta-S100 and NSE occurred in the absence of any detectable effect on intracellular LDH activity, and independently of total cellular protein inhibition. The sensitive and excess responses of the neurospecific endpoints relative to that of the non-specific endpoints may reflect the specific neurotoxic insults of the heavy metals on the cultured cells.(ABSTRACT TRUNCATED AT 250 WORDS)

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Section: Methodsmentioning

confidence: 99%

Neuron and glial cell marker proteins as indicators of heavy metal-induced neurotoxicity in neuroblastoma and glioma cell lines

et al. 1993

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“…Cell mortality was studied by cell flow cytometry using fluorochrome propidium iodide 6 in isolated cells exposed to different pHs (7.4, 5, 2) for 15 min. In cultured cells, mortality was assessed after 24 h of acid exposure (pH 7, 6.5, 5) by measuring the LDH in cell lysates by a colourimetric method with pyruvate and NADH (500Sigma, St Louis, MO) 11 . In this case, intracellular LDH is a better indicator of cell mortality, since the determination of LDH in an acidified culture medium may affect the enzymatic reaction.…”

Section: Methodsmentioning

confidence: 99%

Effects of extracellular pH on restitution and proliferation of rabbit oesophageal epithelial cells

Jiménez

Lanas

Piazuelo

et al. 1999

Aliment Pharmacol Ther

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Background and Aims : Exposure to luminal acid induces mucosal damage in the oesophagus. The extent and degree of interaction of acid with wound repair mechanisms of the oesophagus have not been explored. Methods : Rabbit oesophageal cells were isolated, cultured and exposed to different extracellular pHs for several periods of time. Cell mortality was studied in both isolated and primary cultures of these cells by cell flow cytometry using fluorochrome propidium iodide and LDH release. Subconfluent cell cultures were used to study proliferative responses determined by [3H]‐thymidine incorporation into DNA. Restitution was studied in confluent monolayers wounded by mechanical denudation. Results : Isolated cells were resistant to acid when exposed to pH > 2 for 15 min. Cell proliferation increased with small pH decreases (7–6.5) if the period of exposure was less than 6 h, but was inhibited with lower pHs and/or longer periods of acid exposure. However, restitution was gradually inhibited with further pH decreases (6.5–3) and time of acid exposure (0.5–24 h). Conclusions : The wound repair activities of oesophageal epithelial cells are deeply affected by even small decreases in pH. These findings may help to explain the need for profound acid inhibition in acid‐related reflux oesophagitis.

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“…The damage of hepatocytes was assessed by measuring the percentage of lactate dehydrogenase (LDH) released into the medium [12].…”

Section: Assessment O F Hepatocytes After Isolationmentioning

confidence: 99%

Comparison of Rat Liver Preservation with EurocoUins and a Modified University of Wisconsin Solution: Transplantation and Isolation of Hepatocytes for Culture

Michel

Vons²,

Hillaire³

et al. 1990

Eur Surg Res

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The efficiency of EurocoUins or modified University of Wisconsin (UW) solution (MUW) in preserving rat livers was compared. After cold storage with one of the solutions, the livers were transplanted or perfused by collagenase for isolation of hepatocytes. Five of the 6 rats receiving a graft preserved with MUW versus none of the 6 rats receiving a graft preserved with EurocoUins solution survived 24 h or more. A significantly greater number of hepatocytes were isolated from livers preserved with MUW than from livers preserved with EurocoUins solution. This suggests a better reperfusion of MUW-preserved livers by collagenase resulting from less endothelial injury. LDH release by cultured hepatocytes, ketone body production and stimulation by glucagon were not significantly different between the two groups. These results confirm the superiority of MUW solution over EurocoUins in preserving liver grafts. They suggest that the advantage of MUW solution results from better protection of vascular endothelium rather than of hepatocytes.

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Intracellular lactate dehydrogenase concentration as an index of cytotoxicity in rat hepatocyte primary culture

Cited by 36 publications

References 19 publications

Neuron and glial cell marker proteins as indicators of heavy metal-induced neurotoxicity in neuroblastoma and glioma cell lines

Neuron and glial cell marker proteins as indicators of heavy metal-induced neurotoxicity in neuroblastoma and glioma cell lines

Effects of extracellular pH on restitution and proliferation of rabbit oesophageal epithelial cells

Comparison of Rat Liver Preservation with EurocoUins and a Modified University of Wisconsin Solution: Transplantation and Isolation of Hepatocytes for Culture

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