• Tuberculosis and Respiratory Diseases
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• v.45 no.6
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• pp.1236-1251
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• 1998

Background : TNF-$\alpha$ appears to be a central mediator of the host response to sepsis. While TNF-$\alpha$ is mainly considered a proinflammatory cytokine, it can also act as a direct cytotoxic cytokine. However, there are not so many studies about the relationship bet ween TNF-$\alpha$ level and lung injury severity in ALI, particularly regarding the case of ALI caused by direct lung injury such as diffuse pulmonary infection. Recently, a natural defense mechanism, known as the stress response or the heat shock response, has been reported in cellular or tissue injury reaction. There are a number of reports examining the protective role of pre-induced heat stress proteins on subsequent LPS-induced TNF-$\alpha$ release from monocyte or macrophage and also on subsequent LPS-induced ALI in animals. However it is not well established whether the stress protein synthesis such as HSP can be induced from rat alveolar macrophages by in vitro or in vivo LPS stimulation. Methods : We measured the level of TNF-$\alpha$, the percentage of inflammatory cells in bronchoalveolar lavage fluid, protein synthesis in alveolar macrophages isolated from rats at 1, 2, 3, 4, 6, 12, and 24 hours after intratracheal LPS instillation. We performed histologic examination and also obtained histologic lung injury index score in lungs from other rats at 1, 2, 3, 4, 6, 12, 24 h after intratracheal LPS instillation. Isolated non-stimulated macrophages were incubated for 2 h with different concentration of LPS (0, 1, 10, 100 ng/ml, 1, or 10 ${\mu}g/ml$). Other non-stimulated macrophages were exposed at $43^{\circ}C$ for 15 min, then returned to at $37^{\circ}C$ in 5% CO2-95% for 1 hour, and then incubated for 2 h with LPS (0, 1, 10, 100ng/ml, 1, or 10 ${\mu}g/ml$). Results : TNF-$\alpha$ levels began to increase significantly at 1 h, reached a peak at 3 h (P<0.0001), began to decrease at 6 h, and returned to control level at 12 h after LPS instillation. The percentage of inflammatory cells (neutrophils and alveolar macrophages) began to change significantly at 2 h, reached a peak at 6 h, began to recover but still showed significant change at 12 h, and showed insignificant change at 24 h after LPS instillation compared with the normal control. After LPS instillation, the score of histologic lung injury index reached a maximum value at 6 h and remained steady for 24 hours. 35 kDa protein band was newly synthesized in alveolar macrophage from 1 hour on for 24 hours after LPS instillation. Inducible heat stress protein 72 was not found in any alveolar macrophages obtained from rats after LPS instillation. TNF-$\alpha$ levels in supernatants of LPS-stimulated macro phages were significantly higher than those of non-stimulated macrophages(p<0.05). Following LPS stimulation, TNF-$\alpha$ levels in supernatants were significantly lower after heat treatment than in those without heat treatment (p<0.05). The inducible heat stress protein 72 was not found at any concentrations of LPS stimulation. Whereas the 35 kDa protein band was exclusively found at dose of LPS of 10 ${\mu}g/ml$. Conclusion : TNF-$\alpha$ has a direct or indirect close relationship with lung injury severity in acute lung injury or acute respiratory distress syndrome. In vivo and in vitro LPS stimulation dose not induce heat stress protein 72 in alveolar macrophages. It is likely that 35 kDa protein, synthesized by alveolar macrophage after LPS instillation, does not have a defense role in acute lung injury.

• Tuberculosis and Respiratory Diseases
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• v.45 no.6
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• pp.1265-1276
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• 1998

Background : Nitric oxide(NO) is an endogenously produced free radical that plays an important role in regulating vascular tone, inhibition of platelet aggregation and white blood cell adhesion to endothelial cells, and host defense against infection. The highly reactive nature of NO with oxygen radicals suggests that it may either promote or reduce oxidant-induced cell injury in several biological pathways. Oxidant injury and interactions between pulmonary vascular endothelium and leukocytes are important in the pathogenesis of acute lung injury, including acute respiratory distress syndrome(ARDS). In ARDS, therapeutic administration of NO is a clinical condition providing exogenous NO in oxidant-induced endothelial injury. The role of exogenous NO from NO donor or the suppression of endogenous NO production was evaluated in oxidant-induced endothelial injury. Method : The oxidant injury in cultured rat lung microvascular endothelial cells(RLMVC) was induced by hydrogen peroxide generated from glucose oxidase(GO). Cell injury was evaluated by $^{51}$chromium($^{51}Cr$) release technique. NO donor, such as S-nitroso-N-acetylpenicillamine(SNAP) or sodium nitroprusside(SNP), was added to the endothelial cells as a source of exogenous NO. Endogenous production of NO was suppressed with N-monomethyl-L-arginine(L-NMMA) which is an NO synthase inhibitor. L-NMMA was also used in increased endogenous NO production induced by combined stimulation with interferon-$\gamma$(INF-$\gamma$), tumor necrosis factor-$\alpha$(TNF-$\alpha$), and lipopolysaccharide(LPS). NO generation from NO donor or from the endothelial cells was evaluated by measuring nitrite concentration. Result : $^{51}Cr$ release was $8.7{\pm}0.5%$ in GO 5 mU/ml, $14.4{\pm}2.9%$ in GO 10 mU/ml, $32.3{\pm}2.9%$ in GO 15 mU/ml, $55.5{\pm}0.3%$ in GO 20 mU/ml and $67.8{\pm}0.9%$ in GO 30 mU/ml ; it was significantly increased in GO 15 mU/ml or higher concentrations when compared with $9.6{\pm}0.7%$ in control(p < 0.05; n=6). L-NMMA(0.5 mM) did not affect the $^{51}Cr$ release by GO. Nitrite concentration was increased to $3.9{\pm}0.3\;{\mu}M$ in culture media of RLMVC treated with INF-$\gamma$ (500 U/ml), TNF-$\alpha$(150 U/ml) and LPS($1\;{\mu}g/ml$) for 24 hours ; it was significantly suppressed by the addition of L-NMMA. The presence of L-NMMA did not affect $^{51}Cr$ release induced by GO in RLMVC pretreated with INF-$\gamma$, TNF-$\alpha$ and LPS. The increase of $^{51}Cr$ release with GO(20 mU/ml) was prevented completely by adding 100 ${\mu}M$ SNAP. But the add of SNP, potassium ferrocyanate or potassium ferricyanate did not protect the oxidant injury. Nitrite accumulation was $23{\pm}1.0\;{\mu}M$ from 100 ${\mu}M$ SNAP at 4 hours in phenol red free Hanks' balanced salt solution. But nitrite was not detectable from SNP upto 1 mM The presence of SNAP did not affect the time dependent generation of hydrogen peroxide by GO in phenol red free Hanks' balanced salt solution. Conclusion : Hydrogen peroxide generated by GO causes oxidant injury in RLMVC. Exogenous NO from NO donor prevents oxidant injury, and the protective effect may be related to the ability to release NO. These results suggest that the exogenous NO may be protective on oxidant injury to the endothelium.

• Tuberculosis and Respiratory Diseases
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• v.56 no.1
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• pp.51-66
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• 2004

Objectives : The matrix metalloproteinases (MMPs) that participate in the extracellular matrix metabolism play a important role in the progression of pulmonary fibrosis. The effects of the MMPs are regulated by several factors including Th-1 cytokines, $interferon-{\gamma}$ ($IFN-{\gamma}$). Up to now, $IFN-{\gamma}$ is known to inhibit pulmonary fibrosis, but little is known regarding the exact effect of $IFN-{\gamma}$ on the regulation of the MMPs. This study investigated the effects of $interferon-{\gamma}$ on the pulmonary fibrosis and the expression of the lung MMP-2,-9, TIMP-1,-2, and Th-2 cytokines in aa rat model of bleomycin induced pulmonary fibrosis. Materials and methods : Male, specific pathogen-free Sprague-Dawley rats were subjected to an intratracheal bleomycin instillation. The rats were randomized to a saline control, a bleomycin treated, and a bleomycin+$IFN-{\gamma}$ treated group. The bleomycin+$IFN-{\gamma}$ treated group was subjected to an intramuscular injection of $IFN-{\gamma}$ for 14 days. At 3, 7, 14, and 28 days after the bleomycin instillation, the rats were sacrificed and the lungs were harvested. In order to evaluate the effects of the $IFN-{\gamma}$ on lung fibrosis and inflammation, the lung hydroxyproline content, inflammation and fibrosis score were measured. Western blotting, zymography and reverse zymography were performed at 3, 7, 14, 28 days after bleomycin instillation in order to evaluate the MMP-2,-9, and TIMP-1,-2 expression level. ELISA was performed to determine the IL-4 and IL-13 level in a lung homogenate. Results : 1. 7 days after bleomycin instillation, inflammatory changes were more severe in the bleomycin+$IFN-{\gamma}$ group than the bleomycin group (bleomycin group : bleomycin+$IFN-{\gamma}$ group=$2.08{\pm}0.15:2.74{\pm}0.29$, P<0.05), but 28 days after bleomycin instillation, lung fibrosis was significantly reduced as a result of the $IFN-{\gamma}$ treatment (bleomycin group : bleomycin+$IFN-{\gamma}$ group=$3.94{\pm}0.43:2.64{\pm}0.13$, P<0.05). 2. 28 days after bleomycin instillation, the lung hydroxyproline content was significantly reduced as a result of $IFN-{\gamma}$ treatment (bleomycin group : bleomycin+$IFN-{\gamma}$ group=$294.04{\pm}31.73{\mu}g/g:194.92{\pm}15.51{\mu}g/g$, P<0.05). 3. Western blotting showed that the MMP-2 level was increased as a result of the bleomycin instillation and highest in the 14 days after bleomycin instillation. 4. In zymography, the active forms of MMP-2 were significantly increased as a result of the $IFN-{\gamma}$ treatment 3 days after the bleomycin instillation, bleomycin+$IFN-{\gamma}$ group (bleomycin group : bleomycin+$IFN-{\gamma}$ group=$209.63{\pm}7.60%:407.66{\pm}85.34%$, P<0.05), but 14 days after the bleomycin instillation, the active forms of MMP-2 were significantly reduced as a result of the $IFN-{\gamma}$ treatment (bleomycin group : bleomycin+$IFN-{\gamma}$ group=$159.36{\pm}20.93%:97.23{\pm}12.50%$, P<0.05). 5. The IL-4 levels were lower in the bleomycin and bleomycin+$IFN-{\gamma}$ groups but this was not significant, and the IL-13 levels showed no difference between the experiment groups. Conclusion : The author found that lung inflammation was increased in the early period but the pulmonary fibrosis was inhibited in the late stage as a result of $IFN-{\gamma}$. The inhibition of pulmonary fibrosis by $IFN-{\gamma}$ appeared to be associated with the inhibition of MMP-2 activation by $IFN-{\gamma}$. Further studies on the mechanism of the regulation of MMP-2 activation and the effects of MMP-2 activation on pulmonary fibrosis is warranted in the future.

• Journal of the Korean Society of Food Science and Nutrition
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• v.44 no.4
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• pp.506-515
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• 2015

In this study, the effect of Goheung pomegranate extract on postmenopausal syndrome was evaluated both in vitro and in vivo in ovariectomized Sprague-Dawly (SD) rats. Sixty female SD rats were divided into six groups: sham, sham operation and distilled water; OVX, ovariectomized and distilled water; PE1, ovariectomized and pomegranate concentrate (0.75 mL/twice/d); PE2, ovariectomized and pomegranate concentrate (1.5 mL/twice/d); PE3, ovariectomized and pomegranate concentrate (2.2 mL/twice/d); and CE, ovariectomized and commercial pomegranate concentrate (2.2 mL/twice/d). Percent bone volume (bone volume/tissue volume) and trabecular thickness (Tb.Th) improved in a dose-independent manner in PE1, 2, and 3. Especially, bone mineral density was significantly improved in PE3 (P<0.05) compared to OVX. Pomegranate extract reduced body weight and visceral fat mass. High density lipoprotein cholesterol (HDL-C) level slightly increased in a dose-independent manner in the experimental group. In addition, HDL-C/total cholesterol level of PE3 significantly increased (P<0.05) compared with OVX. These results show that pomegranate concentrate improved blood lipid levels and bone metabolism in ovariectomized rats. Therefore, Goheung pomegranate concentrates are expected to improve cardiovascular and bone-related diseases in menopausal women.

• Clinical and Experimental Reproductive Medicine
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• v.27 no.4
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• pp.373-380
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• 2000

Objective: The purpose of this study was to evaluate effects of goat milk and fermented goat milk on reproductive function and stamina of male rodent. Methods: Experiment I: Male ICR mouse was divided into four groups. Group 1 none-treated control; Group 2 received saline; Group 3 received cow milk 10 ml/kg per day for 15 days; Group 4 received goat milk 10 ml/kg per day for 15 days. The cauda epididymal sperm motility and testicular sperm production were investigated. Experiment II: Male SD rat was divided into three groups. Group 1 received saline; Group 2 received goat milk 10 ml/kg per day for 28 days; Group 3 received fermented goat milk 10 ml/kg per day for 28 days. The cauda epididymal sperm motility and testicular sperm production were also investigated. The concentration of testosterone in serum at 1 and 3 weeks after treatment was determined using Immulite 2000 kit. Testes, epididymis, prostate, and seminal vesicle were weighed. Experiment III: Male ICR mouse was divided into four groups. Group 1 none-treated control; Group 2 received saline; Group 3 received goat milk 10 ml/kg per day for 4 weeks; Group 4 received fermented goat milk 10 ml/kg per day for 4 weeks. After treatment, the mouse was forced to swim to test for stamina. Results: In Experiment I, the cauda epididymal sperm motility after in vitro culture for 1 or 3 h was significantly (p<0.05) higher in cow milk and goat milk than in the control and saline. There was no significant difference in the cauda epidymal sperm motility between cow and goat milk. The testicular spermatid number was significantly (p<0.01) higher in goat milk (222.8${\times}10^6$) than in the control (108.6), saline (98.2), and cow milk (118.2). In Experiment II, the cauda epididymal sperm motility after in vitro culture for 1 h was significantly (p<0.05) higher in fermented goat milk than in saline and goat milk. There was no significant difference in the cauda epidymal sperm motility between saline and goat milk but goat milk showed slightly higher sperm motility than saline. After in vitro culture for 3 h, the cauda epididymal sperm motility was significantly (p<0.01) higher in fermented goat milk and goat milk than in saline. The testicular spermatid number was significantly (p<0.05) higher in goat milk than in saline, and significantly (p<0.01) higher in fermented goat milk than in saline. And the serum testosterone levels of rats administered with goat milk or fermented goat milk were increased but were no significant difference among three groups. Also the prostate weight was significantly (p<0.05) increased in the goat and fermented goat milk. In Experiment III, the swimming time in the goat milk and fermented goat milk groups was significantly (p<0.01) longer than in the control and saline. There was no significant difference in the swimming time between goat and fermented goat milk but the fermented goat milk showed slightly longer swimming time than the goat milk. Conclusion: The cauda epididymal sperm motility, the testicular spermatid number and stamina were improved when the mice and rats were drunk with goat milk or fermented goat milk.

• Journal of Ginseng Research
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• v.27 no.3
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• pp.103-109
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• 2003

We investigated the effect of ginseng total saponin (GTS) on the regeneration process of experimentally crush injured rat sciatic nerves. The bilateral sciatic nerves of fifty adult male Sprague-Dawley rats were compressed surgically with a straight hemostat for 30 seconds with 1 mm width. Twenty rats were divided into four groups to test the dose-dependent effect of GTS (0, 50, 100, or 150 mg/kg, i.p.). Saline for vehicle control group or GTS dissolved in saline was administerd for three weeks. After that period of time, the numbers of total myelinated axon and degenerated myelin in the sciatic nerves of bilateral legs were examined and analyzed using image analysis system to confirm a morphological effect of GTS. We found that the most effective concentration of GTS for the regeneration of damaged sciatic nerve was 150 mg/kg. In another set of experiment, thirty rats were divided into two groups as saline-treated vehicle group and GTS-treated group (150 mg/kg, i.p.) for three weeks. Every week we examined the numbers of total myelinated axon and degenerated myelin in the sciatic nerves of bilateral legs using image analysis system to evaluate the effect of GTS on injured nerves. We found that the regeneration of damaged sciatic nerves was facilitated in GTS-treated group compared to saline-treated group until two weeks. However, after that period of time we could not observe the significant difference between saline-treated group and GTS-treated group. These results suggest that GTS is a useful adjuvant therapy for the regeneration of the peripheral nerve injury in short period of treatment.

• Food Science and Preservation
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• v.17 no.3
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• pp.398-404
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• 2010

It is known that onions, or bioactive compounds therein, providehealth benefits. The present study was designed to investigate whether a concentrated onion extract lowered blood lipid levels in rats fed a high-fat diet. Initially, male Sprague-Dawley rats were housed singly in an environment in which temperature and light duration were controlled, and had free access to a nutritionally adequate AIN-93G diet and deionized water. After an acclimatization period, rats were weight-matched and assigned to one of the following five groups: 1) a control group, fed the AIN-93G diet mixed with 10% (w/v) lard and 0.7% (w/v) cholesterol to induce hyperlipidemia (control); 2) three experimental groups, fed the AIN-93G diet mixed with a high-fat source plus concentrated onion extract at three different levels (termed the low-dose, medium-dose, and high-dose groups); and, 3) a placebo group, fed the AIN-93G diet with fats plus the same concentrated extract but devoid of onion-derived material. All five groups freely ingested their respective diets over 6 weeks. At 0, 3, and 6 weeks, blood samples were collected from the orbital sinus following overnight food deprivation. At 6 weeks, livers were collected. Both control and experimental groups continually gained body weight throughout the study. No significant differencein body weight gain was observed among groups. However, the serum concentrations of triglycerides, total cholesterol, and non HDL-cholesterol were significantly reduced by ingestion of concentrated onion extract. Also, the hepatic levels of total lipids and total fatty acids, especially C18:1 (oleic acid), were significantly decreased in rats fed a high level of concentrated onion extract, compared with the control and placebo groups. These results provide clear evidence that ingestion of a concentrated onion extract has a profound inhibitory effect on serum lipid levels in rats fed a high-fat diet. Our findings indicate that a concentrated onion extract may be used to alleviate hyperlipidemia by lowering serum cholesterol and triglyceride levels.

• Journal of Yeungnam Medical Science
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• v.4 no.2
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• pp.113-120
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• 1987

The effect of intravesical formalin instillation as a therapeutic modality for intractable bladder hemorrhage is well known. And despite clear evidence of therapeutic efficacy of intravesical cytotoxic drugs and/or BCG immunotherapy, there have been substantial recurrences during followup after transurethral resection for superficial bladder tumor. If formalin injected at the bed of superficial bladder tumor is able to coagulate and necrotize the tumor, it will be greatly helpful to the patients With recurrent bladder tumor developed during followup. Since this technique is applicable on outpatient basis, an economical as well as a psychological burden of the patients can be reduced considerably. The purpose of this study is to evaluate the effect of submucosal formalin injection on rat bladder wall, 36 healthy adult male Sprague-Dawley rats (weighing 350gm in average) were divided into 3 groups: In Group I (control group), 0.01ml of normal saline was injected submucosally at the left posterolateral wall of the bladder opened under intraperitoneal Nembutal anesthesia ; In Group II and III, 0.01 ml of 10% and 4% formalin, respectively, were administered at the same site as in the Group I, two rats in each group were sacrificed at day 1, 2, and 3, and week 1, 2 and 4 after injection, respectively. Gross and microscopic examination of the cystectomized specimen were done in each group. In the Group II, bladder stones were formed at week I, and in both the Group I and III, stones were seen at week 2 post injection. There was no significant difference III histologic findings of the bladder between the group II and III. Mucosal ulcer and/or prominent mucosal disruption was observed at 24 hours after injection in both Group II and III. Epithelial regeneration began at day 2, and was marked at day 3, and epithelial lining was almost normalized one week after injection. Subepithelial edema, telangiectasia and inflammatory reaction were prominent at 24 hours post formalin injection. Subepithelial edema persisted in moderate degree for 1 week. Telangiectasia and inflammatory reaction were noted for 4 weeks. Mild degree of these findings also appeared In the control group. Fibroblastic proliferation appeared at day 2 and persisted in moderate degree for 4 weeks. There has been no mortality or bladder perforation. These results suggest that clinical application of this technique is feasible for the selected cases of recurrent, solitary superficial bladder tumor. However, optimal dosage of formalin in relation to the size of the lesion remains to be investigated.

• Journal of Life Science
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• v.21 no.10
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• pp.1415-1421
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• 2011

The purpose of this study was to identify the effects of an L-arginine supplementation and regular exercise training on NF-${\kappa}B$, TNF-${\alpha}$, iNOS, Cav-1, eNOS and Ang II in the aortas of D-galactose (D-gal) induced aging rats. The male Strague-Dawley rats were treated with a D-galactose aging inducing agent; the D-gal injection (50 mg/kg) was given intraperitoneally for 12 wk. Experimental groups were divided into five groups: (1) Young control group (Y-Con, n=8), (2) Aging control group (A-Con, n=8), (3) Aging exercise group (A-Ex, n=8), (4) Aging exercise group with L-arginine supplementation group (A-Ex+A, n=8), and (5) Aging with L-arginine supplementation group (A-A, n=8). The exercise consisted of running on a treadmill for 60 min/day at 20 m/min for 6 day/wk, at 0% gradient for 12 wk. The L-arginine supplementation was given orally at a dose of 150 mg/kg/day for 12 wk. The findings of this study were as follows: 1. NF-${\kappa}B$, TNF-${\alpha}$, iNOS, Cav-1 and Ang II proteins in the aortas of D-gal induced rats were significantly increased, however, L-arginine supplementation and regular exercise resulted in a significant inhibition in the expression of NF-${\kappa}B$, TNF-${\alpha}$, iNOS, Cav-1 and Ang II proteins. 2. eNOS protein in the aortas of D-gal induced rats was significantly decreased, however, L-arginine supplementation and regular exercise resulted in a significant increase in the expression of eNOS proteins. In conclusion, the findings of the present study reveal that L-arginine supplementation alone or regular exercise alone or in combination with L-arginine supplementation for 12 wk increases anti-inflammatory effects by decreasing NF-${\kappa}B$, TNF-${\alpha}$, and iNOS protein expressions within the aortic tissue. In addition, L-arginine supplementation alone or regular exercise alone or in combination with L-arginine supplementation may prevent endothelial function by up-regulation of eNOS protein in the aortas of D-gal induced aging rats.

• Journal of Life Science
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• v.22 no.4
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• pp.516-523
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• 2012

The purpose of this study was to investigate the effect of exercise and/or L-arginine on abdominal fat, IGF-1 on GH/IGF-1 axis, fibrinogen, and PAI-1 in aged and obese rats. Male Sprague-Dawley rats were treated with a D-galactose aging inducing agent (50 mg/kg) given intraperitoneally for 12 weeks. Thirty-two male Sprague-Dawley rats were treated and divided into four groups: aging-high fat diet group (AG+HF), AG+HF with L-arginine intake group (AG+LA), AG+HF with exercise group (AG+EX), and AG+EX with L-arginine intake group (AG+LA+EX). The experimental rats underwent treadmill training (60 min/day, 6 days/week at 0% gradient) for 12 weeks. L-arginine was given orally (150 mg/kg/day) for 12 weeks. After the experiment, blood was collected from the left ventricle and abdominal fat was extracted. The results showed that GH was significantly increased in AG+EX and AG+AL+EX. IGF-1 was significantly increased in both the AG+AL+EX and AG+EX group ($p$<0.05), while fibrinogen and PAI-1 were not significantly different among the groups. Abdominal fat was significantly decreased in the AG+LA, AG+EX, and AG+LA+EX groups ($p$<0.05) compared with the AG+HF group. In conclusion, this study suggests that exercise alone or L-arginine alone or a combination not only increases the GH and IGF-1 concentration, but also decreases the abdominal fat mass.