• Korean Journal of Veterinary Research
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• v.61 no.3
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• pp.22.1-22.7
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• 2021

Fenbendazole (FBZ) is a commonly used anthelmintics in veterinary medicine that has recently been found to have anticancer effects in humans. On the other hand, few studies have examined the anti-inflammatory effects of FBZ, and its mechanism is unknown. In this study, mouse bone marrow cells (BMs) were treated with lipopolysaccharide (LPS), a representative inflammation-inducing substance, to generate a situation similar to osteomyelitis in vitro. The effect of FBZ on inflammatory BMs was examined by measuring the metabolic activity, surface marker expression, cell nuclear morphology, and mitochondrial membrane potential (MMP) of BMs. FBZ decreased the metabolic activity and MMP of LPS-treated BMs. Annexin V-fluorescein isothiocyanate/propidium iodide staining and Hoechst 33342 staining showed that FBZ reduced the number of viable cells and induced the cell death of inflammatory BMs. In addition, FBZ reduced the proportion of granulocytes more than B lymphocytes in LPS-treated BMs. Overall, FBZ induces cell death by destabilizing the MMP of LPS-induced inflammatory BMs. In addition to anthelmintic and anticancer agent, FBZ can play a role as an anti-inflammatory agent.

• Biomolecules & Therapeutics
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• v.20 no.5
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• pp.492-498
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• 2012

Phytochemicals have been known to exhibit potent antioxidant activity. This study examined cytoprotective effects of phytochemicals including quercetin, catechin, caffeic acid, and phytic acid against oxidative damage in SK-Hep-1 cells induced by the oxidative and non-oxidative metabolism of ethanol. Exposure of the cells to excess ethanol resulted in a significant increase in cytotoxicity, reactive oxygen species (ROS) production, lipid hydroperoxide (LPO), and antioxidant enzyme activity. Excess ethanol also caused a reduction in mitochondrial membrane potential (MMP) and the quantity of reduced glutathione (GSH). Co-treatment of cells with ethanol and quercetin, catechin, caffeic acid and phytic acid significantly inhibited oxidative ethanol metabolism-induced cytotoxicity by blocking ROS production. When the cells were treated with ethanol after pretreatment of 4-methylpyrazole (4-MP), increased cytotoxicity, ROS production, antioxidant enzyme activity, and loss of MMP were observed. The addition of quercetin, catechin, caffeic acid and phytic acid to these cells showed suppression of non-oxidative ethanol metabolism-induced cytotoxicity, similar to oxidative ethanol metabolism. These results suggest that quercetin, catechin, caffeic acid and phytic acid have protective effects against ethanol metabolism-induced oxidative insult in SK-Hep-1 cells by blocking ROS production and elevating antioxidant potentials.

• Journal of Applied Biological Chemistry
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• v.57 no.2
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• pp.113-122
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• 2014

A5E is complex of several medicinal herb ethanol extracts. The aim of this study is investigating the anticancer effect for non-small cell lung cancer. The antitumor effects of A5E on NCI-H460 were examined by regulation of cell proliferation, apoptosis, cell cycle arrest, mitochondrial membrane potential (${\Delta}{\Psi}_m$), and apoptosis-related protein. Cell proliferation was measured by MTS assay. Apoptosis induced by A5E was confirmed by Annexin V-fluorescein isothiocyanate (FITC)/Propidium Iodide (PI) staining, and cell cycle arrest was measured by PI staining. NF-${\kappa}B$ translocation was detected by immunofluorescence and MMP (${\Delta}{\Psi}_m$) was measured by JC-1 staining. The expression of extrinsic pathway molecules such as FasL and FADD were elevated, and procaspase-8 was processed by A5E. In addition, intrinsic pathway related molecules were altered. The Bcl-2 and Bcl-xl levels decreased, Bax increased, and cytochrome C was released. In addition, the mitochondrial membrane potential collapsed, and caspase-3 and poly-(ADP-ribose) polymerase were processed by A5E. Moreover, A5E affected the cellular survival pathway involving phosphatidylinositol 3-kinase (PI3K)/Akt and NF-${\kappa}B$. PI3K and Akt were downregulated, also NF-${\kappa}B$ expression was decreased, and nuclear translocalization was inhibited by A5E. These results suggested that A5E delays proliferation, inhibit cell cycle progression and induce apoptosis in human lung cancer cell. We conclude that A5E is a potential anticancer agent for human lung carcinoma.

• Herbal Formula Science
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• v.26 no.3
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• pp.251-259
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• 2018

Objective : Kurarinone is one of the flavonoids isolated from Sophorae Radix with various biological activities including anti-microbial effect. In this study, we investigated the effects of Kurarinone on tert-butyl hydroperoxide (tBHP)-induced oxidative stress finally leading to apoptosis in human hepatoma cell line HepG2. Methods : To determine the effects on cell viability, the cells were exposed to tBHP ($100{\mu}mol/l$) after pretreatment with kurarinone (0.5 and $1{\mu}g/ml$). Cell viability was measured by MTT assay. To reveal the possible mechanism of cytoprotectivity of kurarinone, levels of reactive oxygen species, intracellular glutathione, mitochondrial membrane potential, and expression of caspase were examined. Results : tBHP-induced cell death was due to oxidative stress and the resulting apoptosis. Kurarinone dose-dependently protected cells from apoptosis when determined by MTT and TUNEL assay. Consistent with this observation, decreased expression of pro-caspase 3/9 protein by tBHP was restored by kurarinone. Kurarinone also showed anti-oxidative effects by inhibiting generation of ROS and depletion of GSH in tBHP-stimulated HepG2 cells. In addition, kurarinone significantly recovered disruption of mitochondrial membrane potential (MMP) as a start sign of hepatic apoptosis induced by oxidative stress. Conclusion : From these results, it was concluded that kurarinone protected tBHP-induced hepatotoxicity with anti-oxidative and anti-apoptotic activities. Our results suggest that kurarinone might be beneficial to hepatic disorders caused by oxidative stress.

• Journal of Animal Reproduction and Biotechnology
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• v.37 no.2
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• pp.113-120
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• 2022

Sperm cryopreservation is a fundamental process for the long-term conservation of livestock genetic resources. Yet, the packaging method has been shown, among other factors, to affect the frozen-thawed (FT) sperm quality. This study aimed to develop a new mini-straw for sperm cryopreservation. In addition, the kinematic patterns, viability, acrosome integrity, and mitochondrial membrane potential (MMP) of boar spermatozoa frozen in the developed 0.25 mL straw, 0.25 mL (minitube, Germany), or 0.5 mL (IMV technologies, France) straws were assessed. Post-thaw kinematic parameters were not different (experiment 1: total motility (33.89%, 32.42%), progressive motility (19.13%, 19.09%), curvilinear velocity (42.32, 42.86), and average path velocity (33.40, 33.62) for minitube and the developed straws, respectively. Further, the viability (38.56%, 34.03%), acrosome integrity (53.38%, 48.88%), MMP (42.32%, 36.71%) of spermatozoa frozen using both straw were not differ statistically (p > 0.05). In experiment two, the quality parameters for semen frozen in the developed straw were compared with the 0.5 mL IMV straw. The total motility (41.26%, 39.1%), progressive motility (24.62%, 23.25%), curvilinear velocity (46.44, 48.25), and average path velocity (37.98, 39.12), respectively, for IMV and the developed straw, did not differ statistically. Additionally, there was no significant difference in the viability (39.60%, 33.17%), acrosome integrity (46.23%, 43.23%), and MMP (39.66, 32.51) for IMV and the developed straw, respectively. These results validate the safety and efficiency of the developed straw and highlight its great potential for clinical application. Moreover, both 0.25 mL and 0.5 mL straws fit the present protocol for cryopreservation of boar spermatozoa.

• The Korea Journal of Herbology
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• v.24 no.4
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• pp.77-86
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• 2009

Objectives : The proliferation and migration of human aortic smooth muscle cells (HASMC) in response to activation by various stimuli plays a critical role in the initiation and development of atherosclerosis. This study was conducted to examine the effects of Nelumbo nucifera leaves (NNL) on the proliferation and migration of HASMC. Additionally, the mechanisms involved in any observed effects were also evaluated. Methods : Apoptotic cells were measured by staining with FITC-labeled annexin V, followed by flow cytometric analysis. The expression level of apoptosis related proteins was confirmed by western blot. And MMP-9 activity was measured by gelatin zymography and MMP-9 expression was measured by ELISA Results : NNL completely inhibited the proliferation of HASMC via induction of the expression of apoptotic proteins including annexin V, cleaved poly ADP-ribose polymerase (PARP), and caspase-3 and -8. NNL treatment resulted in the release of cytochrome c into cytosol, a loss of mitochondrial membrane potential, a decrease in Bcl-2 and Bcl-xL and an increase in Bax expression. NNL also blocked HASMC migration via suppression of MMP-9. Conclusions : Taken together, these results indicate that NNL has the potential for use as an anti-artherosclerosis agent.

• The Journal of Korean Medicine
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• v.24 no.2
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• pp.159-165
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• 2003

Objectives : Coptidis rhizoma (CR) is an oriental medicine that has been used in many traditional prescriptions against diabetes mellitus in Korea for centuries. Our purpose was to determine the protective effect and its action mechanism of CR on the cytotoxicity of pancreatic -cell line (RINm5F cell). Methods : In this experiment, we used methods such as MTT assay for detection of cytotoxicity, DNA fragmentation assay for detection of apoptotic cell death, LDH activity assay for detection of necrotic cell death, and measurement of $DiOC_{6}$ (3) retention for detection of mitochondrial membrane potential (MMP). Background : Nitric oxide (NO) is believed to playa key role in the process of pancreatic -cell destruction leading to insulin-dependent diabetes mellitus (IDDM). Results : Exposure of RINm5F cells to chemical NO donor such as S-nitroso-N-acetylpenicillamine (SNAP) induced cytotoxic events such as DNA fragmentation and lactate dehydrogenase (LDH) release into medium. However, pretreatment of RINm5F cells with CR extract ($10~50{\mu\textrm{g}}/ml$) for 3 hours prevented SNAP-induced DNA fragmentation and LDH release into medium through the inhibition of MMP disruption. Conclusions : These results suggest that CR may be a candidate for a therapeutic or preventing agent against IDDM.

• Clinical and Experimental Reproductive Medicine
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• v.51 no.1
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• pp.20-27
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• 2024

Objective: While sperm freezing (cryopreservation) is an effective method for preserving fertility, it can potentially harm the structure and function of sperm due to an increase in the production of reactive oxygen species. This study aimed to assess the impact of zinc oxide nanoparticles (ZnONPs) and selenium oxide nanoparticles (SeONPs) on various sperm functional parameters, including motility, plasma membrane integrity (PMI), mitochondrial membrane potential (MMP), acrosome membrane integrity (ACi), and malondialdehyde (MDA) levels. Methods: Semen samples were collected from 20 Albino Wistar rats. These samples were then divided into six groups: fresh, cryopreservation control, and groups supplemented with SeONPs (1, 2, 5 ㎍/mL) and ZnONPs (0.1, 1, 10 ㎍/mL). Results: Statistical analysis revealed that all concentrations of SeONPs increased total motility and progressive reduction of MDA levels compared to the cryopreservation control group (p<0.05). However, supplementation with ZnONPs did not affect these parameters (p>0.05). Conversely, supplements of 1 and 2 ㎍/mL SeONPs and 1 ㎍/mL ZnONPs contributed to the improvement of PMI and ACi (p<0.05). Yet, no significant change was observed in MMP with any concentration of SeONPs and ZnONPs compared to the cryopreservation control group (p>0.05). Conclusion: The findings suggest that optimal concentrations of SeONPs may enhance sperm parameters during the freezing process.

• Animal Bioscience
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• v.37 no.4
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• pp.640-654
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• 2024

Objective: The purpose of this study was to explore the effect of sodium salicylate (SS) on semen preservation and metabolic regulation in goats. Methods: Under the condition of low temperature, SS was added to goat semen diluent to detect goat sperm motility, plasma membrane, acrosome, antioxidant capacity, mitochondrial membrane potential (MMP) and metabonomics. Results: The results show that at the 8th day of low-temperature storage, the sperm motility of the 20 μM SS group was 66.64%, and the integrity rates of the plasma membrane and acrosome were both above 60%, significantly higher than those of the other groups. The activities of catalase and superoxide dismutase in the sperm of the 20 μM SS group were significantly higher than those of the control group, the contents of reactive oxygen species and malondialdehyde were significantly lower than those in the control group, the MMP was significantly higher than that in the control group, and the contents of Ca²⁺ and total cholesterol were significantly higher than those in the control group. Through metabonomics analysis, there were significant metabolic differences between the control group and the 20 μM SS group. Twenty of the most significant metabolic markers were screened, mainly involving five metabolic pathways, of which nicotinic acid and nicotinamide metabolic pathways were the most significant. Conclusion: The results indicate that SS can effectively improve the low-temperature preservation quality of goat sperm.

• Parasites, Hosts and Diseases
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• v.48 no.1
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• pp.1-7
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• 2010

Neutrophils play an important role in the human immune system for protection against such microorganisms as a protozoan parasite, Trichomonas vaginalis; however, the precise role of neutrophils in the pathogenesis of trichomoniasis is still unknown. Moreover, it is thought that trichomonal lysates and excretory-secretory products (ESP), as well as live T. vaginalis, could possibly interact with neutrophils in local tissues, including areas of inflammation induced by T. vaginalis in humans. The aim of this study was to investigate the influence of T. vaginalis lysate on the fate of neutrophils. We found that T. vaginalis lysate inhibits apoptosis of human neutrophils as revealed by Giemsa stain. Less altered mitochondrial membrane potential (MMP) and surface CD16 receptor expression also supported the idea that neutrophil apoptosis is delayed after T. vaginalis lysate stimulation. In contrast, ESP stimulated-neutrophils were similar in apoptotic features of untreated neutrophils. Maintained caspase-3 and myeloid cell leukemia-1 (Mcl-1) in neutrophils co-cultured with trichomonad lysate suggest that an intrinsic mitochondrial pathway of apoptosis was involved in T. vaginalis lysate-induced delayed neutrophil apoptosis; this phenomenon may contribute to local inflammation in trichomoniasis.