• Tuberculosis and Respiratory Diseases
• /
• v.41 no.3
• /
• pp.215-221
• /
• 1994

Background: It is well known that oxygen free radicals(OFR) play a vital role in the various type of acute lung injury. Among various antioxidant defense mechanisms, the superoxide dismutases(SOD) are thought to be the first line of antioxidant defense by catalyzing the dismutation of two superoxide radicals to yield hydrogen peroxide and oxygen. Eukaryotic cells contain two types of intracellular SOD : cytosolic, dimeric copper/zinc- containing enzyme(CuZnSOD) and mitochondrial, tetrameric manganese-containing enzyme(MnSOD). The purpose of this study is to evaluate the time-dependent gene expression of MnSOD and CuZnSOD in the endotoxin-treated rats, and to compare with the manifestations of LPS-induced acute lung injury in rats. Methods: Total RNA from rat lung was isolated using single step phenol extraction 0, 1, 2, 4, 6, 12, 18, 24 hours after E. coli endotoxin injection(n=3, respectively). RNA was separated by formaldehyde-containing 1.2% agarose gels elctrophoresis, transblotted, baked, prehybridized, and hybridized with $^{32}P$-labeled cDNA probes for rat MnSOD and CuZnSOD, which were kindly donated by Dr. Ho(Duke University, Durham, NC, USA). The probes were labeled by nick translation. Blots were washed and autoradiography were quantitated using laser densitometry. Equivalent amounts of total RNA/gel were assessed by monitoring 28S and 18S rRNA. Results: Endotoxin caused a rise in steady-state MnSOD mRNA levels by 4h with peak mRNA accumulation by 6h. Continued MnSOD mRNA expression was observed at 12h. CuZnSOD mRNA expression was observed from 1h to 24h with peak levels by 18h. Conclusion: These results suggest that SOD palys an important defensive role in the endotoxin-induced acute lung injury in rats.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.26 no.2
• /
• pp.186-192
• /
• 1997

Increased activities of phase 2 enzymes including quinone reductase(QR) have been reported to be associated with protection of animals from neoplastic, mutagenic, and other toxic effects of many carcinogens. In previous study, we found that methanol extract of roasted and defatted perilla meal induced the activity of quinone reductase, an anticarcinogenic marker enzyme, in murine hepalc1c7 cells. Current study showed that unroasted perilla had a limited QR-inducing activity, suggesting that roasting cause the generation of active component(s). Thus we hypothesized that QR inducer in perilla might be covalently linked to sugar moiety and released during roasting process. Methanol extract of defatted raw perilla was subject to acid treatment in order to hydrolyze the potential sugar moiety. Prolonged hydrolysis of methanol extract of defatted raw perilla at $98{\sim}100^{\circ}C$ increased the ability to induce cytosolic QR activity of hepalclc7 cells. Furthermore roasting at 180 and $200^{\circ}C$ resulted in significant induction of QR activity. The result strongly support the idea that QR inducer(s) is present in bound form in raw perilla and released during roasting. Cellular QR activity was induced proportionately with the increase of concentration of methanol extract of roasted perilla. The induction of QR by defatted perilla was also examined in the cytosols of liver, small intestine, stomach, lung and kidney of male ICR mice. Induction patterns showed specificity with respect to target tissue and roasting of perilla. Unroasted perilla meal (defatted) significantly induced QR in liver and lung, while roasted perilla meal induced QR in liver and stomach. The observation that raw perilla showed similar QR induction patterns to roasted perilla is consistent with our proposal that QR inducer(s) is present in bound form and released by physical and chemical treatments as digestive or microbial enzymes could release the inducers from inactive glycoside forms in gastrointestinal tract of mice. In conclusion, perilla could exert protective effect against chemically induced carcinogenesis by inducing phase 2 enzymes in biological systems regardless of chemical and physical process such as roasting.

• Journal of Life Science
• /
• v.20 no.3
• /
• pp.444-452
• /
• 2010

The purpose of this study was to investigate the effects of treadmill exercise on $A{\beta}$-42, cytochrome c, SOD-1, 2 and Sirt-3 protein expressions in brain cytosol and mitochondria in mutant (N141I) presenilin-2 transgenic mice with Alzheimer's disease (AD). The mice were divided into four groups (Non-Tg-sedentary, n=5; Non-Tg treadmill exercise, n=5; Tg-sedentary, n=5; Tg treadmill exercise, n=5). To evaluate the neuroprotective effect of treadmill exercise, Non-Tg and Tg mice were subjected to exercise training on a treadmill for 12 wk, after which their brain cytosol and mitochondria were evaluated to determine whether any changes in the cognitive performance, $A{\beta}$-42 protein, cytochrome c protein, anti-oxidant enzymes (SOD-1, SOD-2) and Sirt-3 protein had occurred. The results indicated that treadmill exercise resulted in amelioration in cognitive deficits of Tg mice. In addition, the expressions of mitochondrial $A{\beta}$-42 and cytosolic cytochrome c protein were decreased in the brains of Tg mice after treadmill exercise, whereas antioxidant enzymes, SOD-l and SOD-2 were significantly increased in response to treadmill exercise. Furthermore, treadmill exercise significantly increased the expression of Sirt-3 protein in Non-Tg and Tg mice. Taken together, these results suggest that treadmill exercise is a simple behavioral intervention which can sufficiently improve cognitive performance and inhibit $A{\beta}$-induced oxidative stress in AD.

• Journal of Life Science
• /
• v.31 no.3
• /
• pp.257-265
• /
• 2021

Heterotrimeric kinesin-2 is a molecular motor protein of the kinesin superfamily (KIF) that moves along a microtubule plus-end directed motor protein. It consists of three different motor subunits (KIF3A, KIF3B, and KIF3C) and a kinesin-associated protein 3 (KAP3) that form a heterotrimeric complex. Heterotrimeric kinesin-2 interacts with many different binding proteins through the cargo-binding domain of the KIF3s. The activity of heterotrimeric kinesin-2 is regulated to ensure that the cargo is directed to the right place at the right time. How this regulation occurs, however, remains in question. To identify the regulatory proteins for heterotrimeric kinesin-2, we performed yeast two-hybrid screening and found a specific interaction with creatine kinase B (CKB), which is the brain isoform of cytosolic creatine kinase enzyme. CKB bound to the cargo-binding domain of KIF3A but did not interact with the KIF3B, KIF5B, or KAP3 in the yeast two-hybrid assay. The carboxyl (C)-terminal region of CKB is essential for the interaction with KIF3A. Another protein kinase, CaMKIIa, interacted with KIF3A, but GSK3a did not interact with KIF3A in the yeast two-hybrid assay. KIF3A interacted with GST-CKB-C but not with GSK-CKB-N or GST alone. When co-expressed in HEK-293T cells, CKB co-localized with KIF3A and co-immunoprecipitated with KIF3A and KIF3B but not KIF5B. These results suggest that the CKB-KIF3A interaction may regulate the cargo transport of heterotrimeric kinesin-2 under energy-compromised conditions in cells.