• Journal of Environmental Health Sciences
• /
• v.26 no.4
• /
• pp.115-121
• /
• 2000

To evaluate the cytotoxicity of cadmium compounds, this study was conducted to measure the in vitro magnetometry, LDH release and cellular apoptosis using alveolar macrophages of hamsters. A series of magnetometric measurements in cadmium-added groups showed a significant dose-dependent decay of the relaxation curves. The LDH release rates showed a dose-dependently increasing tendency as the dose gradually increased. The positive rates of apoptosis were significantly higher in cadmium-added groups than the control groups. Conclusively, the cytotoxicity increased in a dose dependent way as the concentration of cadmium added increased, which reflected in the decay of relaxation curve in magnetometry, and increased LDH release rate and positive rate of apoptosis.

• Journal of Life Science
• /
• v.18 no.2
• /
• pp.264-272
• /
• 2008

The lactate dehydrogenase (EC 1.1.1.27, LDH) isozymes in tissues from Acanthogobius hasta were characterized by biochemical, immunochemical and kinetic methods. The activities of LDH in skeletal muscle and eye tissues were 65.30 and 53.25 units, but LDH activities in heart and liver tissues were very low. LDH/CS (EC 4.1.3.7, citrate synthase) in skeletal muscle was the highest as 22.29. Specific activities of LDH in brain, eye and skeletal muscle were 56.45, 38.04 and 11.0 units/mg, respectively. The LDH isozymes in tissues were separated by polyacrylamide gel electrophoresis after immunoprecipitation with antiserum against $A_4,\;B_4$ eye-specific $C_4$ and liver-specific $C_4$. LDH $AC_4$ isozymes were detected predominantly in skeletal muscle, brain and eye tissues, and $B_4$ isozyme was detected in heart. Anodal eye-specific $C_4$ and cathodal liver-specific $C_4$ were coexpressed in A. hasta. The eye-specific $C_4$ isozyme showed higher activity in eye tissue, but liver-specific $C_4$ isozyme showed lower activity in liver. As a result, one part of molecular structures in $A_4\;and\;C_4,\;A_4\;and\;B_4$, and eye-specific $C_4$ and liver-specific $C_4$ were similar, but in $B_4\;and\;C_4$ were different with each other. Therefore the subunit A may be conservative in evolution, and the evolution of subunit B seems to be faster than that of subunit A. The LDH $A_4$ isozyme of skeletal muscle was purified in the fraction from elution with NAD+ containing buffer of affinity chromatography and eye-specific $C_4$ isozyme was eluted right after $A_4$, so the structure of eye-specific $C_4$ isozyme is similar to $A_4$. And LDH activity remained 35.22-43.47% as a result of the inhibition by pyruvate, the Michaelis-Menten constant values for pyruvate was 0.080-0.098 mM, and Vmax were 153.85 units, 35.09 units in skeletal muscle and eye, respectively. Also the $B_4$ isozyme was the thermo-stablest and $C_4$ was stabler than $A_4$ isozyme. The optimum pH of LDH was 6.5. The results mentioned above indicate that isozymes in tissues showed the properties between LDH $A_4\;and\;B_4$ isozyme as A. hasta was adapted to hypoxic conditions. Also LDH seems to function more effectively under anaerobic condition because LDH in skeletal muscle and eye tissues have high affinity for pyruvate.

• Biomedical Science Letters
• /
• v.6 no.1
• /
• pp.19-28
• /
• 2000

This study was performed to evaluate the activities of glutathione S-transferase (GST) and lactate dehydrogenase (LDH) in Maddin-Darby canine kidney (MDCK) cells infected with virus and/or treated with amantadine. On cell morphological findings, monolayer fractions in MDCK cells infected with virus were exfolated more than 80% in 1 TCID$_{50}$ group and that in 10 TCID$_{50}$ were completely exfolated after 3 days during infectious process. In proportion to the dose of amantadine, activities of GST and LDH of MDCK cells were significantly decreased and those of LDH in medium fraction were more significantly increased compared with control. According to in both dose and time of virus innoculation, activities of GST and LDH in MDCK cells were significantly decreased in 1 and 10 TCID$_{50}$ infected cells after 3 days. LDH activities in infectious medium were remarkably rised at 10 fold. In case of the cell line inoculated with type A 100 TCID$_{50}$ and additionally treated with amantadine, the decreasing rate to the control in activities of GST and LDH was lower than that in those in case of that infected with virus only. These results suggested that virus infection and amantadine treatment may effect the activity of the detoxicating enzyme in the target cells.

• Journal of Physiology & Pathology in Korean Medicine
• /
• v.22 no.1
• /
• pp.115-119
• /
• 2008

This study was done to evaluate the antioxidant effect of Crataegi Fructus (CF) extract on the oxidative stress induced by reactive oxygen species (ROS), The human skin fibroblasts (Detroit 551) were cultured with various concentrations of hydrogen peroxide $(H_2O_2)$. The cytotoxicity of $H_2O_2-induced$ oxidative stress was performed by XTT assay for the cell viability according to the dose- and time-dependent treatment. For the protective effect of CF extract on $H_2O_2-mediated$ oxidative stress, cell viability, lactate dehydroganase (LDH) activity, and ferric thiocyanate (FTC) assay for the inhibitive activity of lipid peroxidation on CF extract were carried out. In this study, $H_2O_2-mediated$ oxidative stress was decreased cell viability dose-, and time-dependent manner and increased LDH activity compared with the control in these cultures. In the protective effect, CF extract increased cell viability and decreased LDH activity on $H_2O_2-mediated$ oxidative stress, especially, CF extract has antioxidant effect by the showing the inhibitive activity of lipid peroxidation by FTC assay. From these results, It is suggested that $H_2O_2-mediated$ oxidative stress was highly toxic, and also, CF extract showed the protective effect on $H_2O_2-mediated$ oxidative stress by showing the increased cell viability, decreased LDH activity and lipid peroxidation inhibition in these cultures.

• Journal of Life Science
• /
• v.20 no.3
• /
• pp.416-423
• /
• 2010

In this study, the properties and gene expression of the lactate dehydrogenase (EC 1.1.1.27, LDH) isozyme were studied in angelfish (Pterophyllum scalare) - known for their adaptation to the low oxygen environment of the tropics - which were acclimated to acute temperature change ($27{\pm}0.5{\rightarrow}18{\pm}0.5^{\circ}C$) and dissolved oxygen (DO) change ($6{\pm}1{\rightarrow}18\;ppm$) for 2 hours. The properties of the LDH isozymes were confirmed in the native-polyacrylamide gel electrophoresis, Western blot analysis and enzyme activity measurement. Liver- and eye-specific Ldh-C gene were expressed in liver, eye and brain tissues. Through Western blot analysis, the LDH $A_4$ isozyme was shown to have a more cathodal mobility relative to the $B_4$ isozyme. In the liver tissue, the LDH $A_4$ isozyme increased with temperature drop while the $B_4$ isozyme decreased. The LDH $A_4$ and $C_4$ isozymes increased with DO increment, while the $B_4$ isozyme decreased. In the eye tissue, the LDH $A_4$ and B4 isozymse increased with temperature drop while the $B_4$ isozyme decreased. The LDH $A_4$ and $B_4$ isozymes increased with DO increment, but the $C_4$ isozyme and isozymes including the subunit C decreased. In the heart tissue, LDH activity increased with DO increment, as well as the LDH $B_4$ isozyme. In the brain tissue, the LDH $A_4$ and $B_4$ isozymes increased with temperature drop. The LDH $B_4$ isozyme increased with DO increment. Accordingly, since the liver- and eye-specific Ldh-C are influenced by changes in DO and the LDH $B_4$ and $C_4$ isozymes are relatively controlled in the liver and eye tissues, the $C_4$ isozyme can be considered to have a lactate oxidase function.

• Biotechnology and Bioprocess Engineering:BBE
• /
• v.9 no.4
• /
• pp.318-322
• /
• 2004

Lactic acid is an environmentally benign organic acid that could be used as a raw material for biodegradable plastics if it can be inexpensively produced by fermentation. Two genes (ldhL and ldhD) encoding the L-(+) and D-(-) lactate dehydrogenases (L-LDH and D-LDH) were cloned from Lactobacillus sp., RKY2, which is a lactic acid hyper-producing bacterium isolated from Kimchi. Open reading frames of ldhL for and ldhD for the L and D-LDH genes were 962 and 998 bp, respectively. Both the L(+)- and D(-)-LDH proteins showed the highest degree of homology with the L- and D-lactate dehydrogenase genes of Lactobacillus plantarum. The conserved residues in the catalytic activity and substrate binding of both LDHs were identified in both enzymes.

• Journal of Life Science
• /
• v.15 no.1 s.68
• /
• pp.80-86
• /
• 2005

To achieve the purpose of this study, forty-eight male Sprague-Dawley rats were assigned to control and three endurance exercise group. 36 rats were forced to exercise according to exercise intensity for 8 weeks and 12 rats were untrained for control group. Cardiac cytosol was extracted from cardiac tissue and cardiac mitochondria was purified from the cytosol. Purified mitochondria were separated into four fraction: inner membrane, outer membrane inter membrane space and matrix. The changes of cytosolic and mitochondrial LDH isozymes activity were measure. Relative activity $(\%)$ of cytosol for low and control group showed the following order of prevalence $AB_3>A_2B_2>B_4>A_3B>A_4$ for moderate and high group : $AB_3>B_4>A_2B_2>A_3B>A_4$. Outer membrane for low group showed $AB_3>B_4>A_2B_2$, for moderate group:$ B_4>AB_3>A_2B_2$, for high and control group: $B_4>A_3B$. Inter membrane space for low, moderate and high group showed $B_4>AB_3>A_2B_2>A_3B>A_4$, for control group: $B_4>A_3B>AB_3>A_2B_2>A_4$. Inner membrane for all group showed $B_4>AB_3>A_2B_2>A_3B>A_4$. Matrix for control, low, moderate and high group showed $B_4>AB_3>A_2B_2>A_3B>A_4$. These results suggest that long term exercise intensity effect on cardiac tissue cytosolic and mitochondrial activity and $A_4,\;A_3B,\;A_2B_2,\;AB_3\;and\;B_4$ isozymes were found entirely in mitochondrial fraction.

• Proceedings of the Ginseng society Conference
• /
• 1990.06a
• /
• pp.58-64
• /
• 1990

The effects of ginseng saponins, G-Rbl and G-Rc on the rat liver LDH A-gene transcnptional activity was investigated during pro-replicative phase of rat liver after partial hepatectomy. Changes in LDH A-mRNA levels in regenerating rat liver after intraperitoneal administrations of G-Rbl of G-Rc were tested by slot blot hybridization methods. The results showed that G-Rbl (1 mg/100g B.W) and G-Rc (1 ma/100g B.W) caused marked increases of LDH A-mRNA contents by respectively 1.9- and 1.5-fold in rat liver at 5·hours after partial hepatectomy. Dose dependent effect of G-Rbl and G-Rc (1-25 mg/100g B.W) on the LDH A-mRNA levels on regenerating rat liver were also analyzed. The maximal in- creases of liver LDH A-mRNA levels were observed with the doses of 1 mg for G-Rbl and 5 mg for G-Rc However, when the administration doses of G-Kbl and G-Rc were increased to 20 mg, G-Rbl caused a marked decrease of LDH A-mRNA level to 61% of those in sham-operated rat liver In contrast, G-Rc slightly decreased the liver LDH A-mRNA contents by 30% as compared to those of the maximum value but still maintained 22% higher LDH A-mRNA levels then those of sham-operated rate liver. On the basis of these experimental results, we conclude that ginseng saponin, G-Rb 1 and G·Rc have stimulatory effect at the lower concentration (1 mg/100g B.W) and inhibitory effect at the higher concentration (20 moi loos 5.W) on the LDH A-gene transcription during regeneration of rat liver, Additionally we also investigated the stimulatory effects of ginsenosides on the protein and DNA synthetic activities in hepatocyte primary cell cultures isolated from regenerating rat liver. Both of G·Rc and -Re increased the synthetic rates of hepatocytes proteins and DNA at the administration doses of 50 ug and 100 ug/3 ml/dish respectively representing 1.3-1.6 fold increases. From these results we postulate that G-Rc and -Re may have a mitogen enhancer activity for the hepatocyte proliferation during rat liver regeneration period. Keywords Inductive effects of ginsenosides, G-Rb, -Rc, and -Re, rat LDH A-gene transcription, the sin thetic rate of proteins and DNA in regeneration rat liver.

• Journal of Ginseng Research
• /
• v.14 no.2
• /
• pp.200-206
• /
• 1990

The effects of ginseng saponins, SRbl and G-Rc on the rat liver LDH A-gene transcriptional activity was investigated during prereplicative phase of rat liver after partial hepatectomy. Changes in LDH A-mRNA levels in regenerating rat liver after intraperitoneal administrations of G-Rbl or 'G-Rc were tested by slot blot hybridization methods. The results showed that G-Rbl (1 mg/100g B.W) and G-Rc (1 mg/100g B.W) caused marked increases of LDH A-mRNA contents by respectively 1.9- and 1.5-fold in rat liver at 5-hours after partial hepatectomy Dose dependent elect of G-Rbl and G-Rc (1-25 mg/ 100g B.W) on the LDH A-mRNA levels on regenerating rat liver were also analyzed. The maximal increases of liver LDH A-mRNA levels were observed with the doses of 1 mg for G-Rbl and 5 mg for G-Rc. However, when the administration doses of G-Rbl and G-Rc were increased to 20 mg, G-Rbl caused a marked decrease of LDH A-mRNA level to 61% of those in sham-operated rat liver. In contrast, G-Rc slightly decreased the liver LDH A-mRNA contents by 30% as compared to those of the maximum value but still maintained 22% higher LDH A-mRNA levels then those of sham-operated rate liver. On the basis of these experimental results, we conclude that ginseng saponin, G-Rbl and G-Rc have stimulatory effect at the lower concentration (1 mg/ 100g B.W) and inhibitory effect at the higher concentration (20 mg/ 100g B.W) on the LDH A-gene transcription during regeneration of rat liver. Additionally we also investigated the stimulatory effects of ginsenosides on the protein and DNA sinthetic activities in hepatocyte primary cell cultures isolated from regenerating rat liver. Both of G-Rc and -Re increased the synthetic rates of hepatocytes proteins and DNA at the administration doses of 50 us and 100 $\mu\textrm{g}$/3 ml/dish respectively representing 1.3-1.6 fold increases. From these results we postulate that G-Rc and -Re may have a mitogen ehincer activity for the hepatocyte proliferation during rat liver regeneration period.

• Biomedical Science Letters
• /
• v.15 no.4
• /
• pp.355-362
• /
• 2009

I evaluated the protective effect of N-methyl-D-aspartate (NMDA)/glycine receptor antagonist, 7-chlorokinurenic acid (CKA) on cultured mouse astrocytes damaged by ischemia-like condition (ILC). The protective effect of CKA was assessed by cell viability, lactate dehydrogenase (LDH) activity, superoxide dismutase (SOD)-like activity and lipid peroxidation. To examine the effect of CKA on the cell apoptosis, the expression and the activity of caspase 3 were assessed by Western blotting. CKA increased the cell viability decreased by ILC. CKA also decreased the LDH activity and antioxidative effects such as SOD-like activity and inhibitory activity of lipid peroxidation. In addition, CKA suppressed the expression of caspase 3 associated with apoptosis, and increased the cell viability by the decrease of caspase 3 activity as like the caspase 3 inhibitor, Av-DVED-MED. From these results, these results suggest that ILS induces cell cytotoxicity in cultured astrocytes and CKA, NMDA/glycine receptor antagonist, is effective on the prevention of the cytotoxicity due to ILS by the antioxidative effect and the inhibition of apoptosis.