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Study of hepatoprotective effect of Haegan-jeon through activation of nuclear factor erythroid 2-related factor 2 and optimization of herbal composition based on molecular mechanism (Nuclear factor erythroid 2-related factor 2 활성화를 통한 해간전(解肝煎)의 간세포 보호 효능 및 분자기전을 활용한 해간전(解肝煎) 구성 약물의 최적화 연구)

  • Kim, Jae Kwang;Jung, Ji Yun;Park, Sang Mi;Park, Chung A;Ku, Sae Kwang;Byun, Sung Hui;Cho, Il Je;Kim, Sang Chan
    • Herbal Formula Science
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    • v.26 no.3
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    • pp.207-221
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    • 2018
  • Objectives : Present study investigated hepatoprotective effect of Haegan-jeon extract (HE) and tried to elucidate molecular mechanism involved. According to molecular mechanism, present study optimized herbal composition of HE (op-HE) and compared in vitro and in vivo hepatoprotective effects of op-HE to HE. Methods : For in vitro experiments, HepG2 cells were exposed to arachidonic acid (AA, $10{\mu}M$) and iron ($5{\mu}M$) for inducing oxidative stress. Cell viability, GSH contents, $H_2O_2$ production, mitochondrial membrane potential, immunoblot and reporter gene assay were performed to investigate cytoprotective effects and responsible molecular mechanisms. For in vivo experiments, hepatoprotective effect of HE and op-HE were assessed on $CCl_4-induced$ liver injury mice model. Results : HE pretreatment prevented AA+iron-mediated hepatocytes apoptosis. In addition, AA+iron-induced mitochondrial dysfunction, $H_2O_2$ production, glutathione depletion were reduced by HE pretreatment. In addition, nuclear factor erythroid 2-related factor 2 (Nrf2) phosphorylation, antioxidant response element (ARE)-driven reporter gene activity, and antioxidant genes expression were increased by HE. Based on reporter gene and MTT assays, we found that op-HE consisting three medicinal herbs also significantly increased transactivation of Nrf2 and reduced the AA+iron-mediated cytotoxicity. Moreover, in $CCl_4-induced$ liver injury mice model, HE-op had an ability to ameliorate $CCl_4-mediated$ increases in serum alanine transferase and aspartate aminotransferase activity, hepatic degeneration, inflammatory cell infiltration, and collagen deposition. Hepatoprotective effects of op-HE were comparable to those of HE. Conclusions : Present study suggests that op-HE as well as HE exhibit hepatoprotective effect against oxidative stress-mediated liver injury via Nrf2 activation.

A Toxicological Study of Young Fronds of Bracken Fern (Pteridium aquilinum var latiusculum) Collected in Kwang Ju Area (한국산 고사리의 독성조사에 관한 연구)

  • Sheo, Hwa-Joong;Lee, Myung-Yul
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.18 no.3
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    • pp.255-264
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    • 1989
  • The results of toxicity test using mice and rats for ethanol extract of Bracken Fern collected in Kwang Ju area were shown as follows ; Up to the dose of 10g per kg of mice administered intraperitoneally there was no lethal toxicity so that it was impossible to calculate the median lethal dose $(LD_{50})$. For the first 7 days experiment all rats administered frond extract grouping in 40mg, 400mg, and 1200mg per kg of rat as the daily oral doses did not show any characterized sign in the weight gain rate, anatomical findings, and biochemical studies. For 3 weeks following the first week the weight gain rates of all test group were reduced to $4.2{\sim}7%$ below the weight gain rate of control. In this period serum GPT, GOT, and Alkaline phosphatase value were increased significantly indicating the symptoms of Bracken Fern poisonings. The pathological findings of all test groups for 28 days showed acute and chronic intestinal lesion and liver damage with steatosis especially in 1200g/kg rat groups. In this experiment the Bracken poisonings appeared slowly in rats of 400mg/kg and 1200mg/kg for two weeks and in rats of 40mg/kg for 3 weeks, showing the symptoms of lowering of weight gain rate, subacute hepatitis, hepatic steatosis and enteritis in 28 days experiment.

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The Effects of Samul-tang-ga-dansam for Wound Healing (사물탕(四物湯) 가(加) 단참(丹參)의 상처 치료에 대한 효과)

  • Eun-Byeol Lee;Hyeon-Ji Kim;Chae-Young Kim;Ji-Su Choi;Chang-Hoon Woo;Young-Jun Kim;Hee-Duk An
    • Journal of Korean Medicine Rehabilitation
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    • v.33 no.2
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    • pp.1-18
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    • 2023
  • Objectives The purpose of this study was to evaluate the antioxidant, anti-inflammatory and wound healing effects of Samul-tang-ga-dansam water extract (SD) in wound-induced mice. Methods The mice were divided into five groups (n=7): the normal group, the control group, the positive control group, the low-dose SD group and the high-dose SD group. The normal group had no wounds and the other groups were wounded on the back with a leather punch. Distilled water was administered to the control group, 200 mg/kg of vitamin E was administered to the positive control group. In the low-dose SD group and the high-dose SD group, 1.23 g/kg and 2.47 g/kg of SD were administered, respectively. Antioxidant and anti-inflammatory protein levels were evaluated using western blot analysis. Skin tissue was analyzed by H&E, Masson's trichrome staining method. Results Oral administration of the SD significantly reduced the visible skin damage and decreased the reactive oxygen species and ONOO- activity of the serum. It significantly increased heme oxygenase-1, superoxide dismutase, catalase, GPx-1/2, Nrf2 and Keap-1 which are antioxidant-related factors in skin tissue and reduced NF-κB p65, inducible nitric oxide synthase, cyclooxygenase-2, tumor necrosis factor α, interleukin (IL)-1β, IL-6 which are inflammation-related factors. Also, SD significantly decreased NOX2, p22phox and p47phox and increased α-smooth muscle actin and COL1A1 protein expression in fibroblasts involved in connective tissue repair. According to histological examination, the thickened epithelial layer was thinned and collagen fibers were increased to accelerate wound healing. Conclusions It is suggested that Samul-tang-ga-dansam has antioxidant and anti-inflammatory effects and promotes wound tissue repair.

The Effect of Dangguijakyak-san on Wound Healing (당귀작약산의 창상 회복에 대한 효과)

  • Yun-Jin Lee;Chang-Hoon Woo;Young-Jun Kim;Hyeon-Ji Kim;Hee-Duk An
    • Journal of Korean Medicine Rehabilitation
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    • v.33 no.3
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    • pp.47-65
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    • 2023
  • Objectives We evaluated the wound healing effects of Dangguijakyak-san (DJ) using C57BL/6 mice that were generated open wound. Methods The study was conducted with seven C57BL/6 mice assigned to each group, divided into the normal group, control group, vitamin E group, DJ low-dose group, DJ high-dose group. We measured total polyphenol, flavonoid contents, the size of the wound, liver function, pro-inflammatory cytokine activity in serum, inflammation-related proteins, adhesion molecules and chemokine proteins, collagen-related proteins in skin tissue and histopathological changes by H&E and Masson's staining. Results DJ treatment significantly reduced the area of the wound compared to the control group. Also, inflammatory cytokines were reduced and the expression of anti-inflammatory-related factors (interleukin-4 [IL-4] and IL-10) was significantly increased in the DJ treatment group. We identified that DJ treatment inhibits both pathways of inflammation, the mitogen-activated protein kinases and nuclear factor-κB pathway. Moreover, the protein expressions of Sirt1 (sirtuin 1), MCP-1 (monocyte chemoattractant protein 1), ICAM-1 (intercellular adhesion molecule 1), and VCAM-1 (vascular cell adhesion molecule 1) were decreased by DJ administration. Also, the expression of α-smooth muscle actin and collagen type I alpha 1, collagen-related proteins, that help skin recovery was significantly increased in the DJ treatment group. Histopathologically, a relatively thin epithelial layer could be observed in the DJ administration group, as well as an increase in fibroblasts and collagen fibers. Conclusions These data suggest that DJ treatment is effective in wound healing, suppressing inflammatory proteins, increasing skin repair factors and improving histopathological changes caused by wounds.

The Effect of Antioxidant Vitamins on Aflatoxin $B_{1}-DNA$ Adduct the Formation in Aflatoxin $B_1$ Administered Mice Liver (항산화비타민이 Aflatoxin $B_1$ 투여 마우스의 간세포에서 Aflatoxin $B_{1}-DNA$ Adduct 형성에 미치는 영향)

  • Park, Seon-Ja;Kang, Sung-Jo;Park, Jung-Hyun;Oh, Sang-Suk;Chung, Duck-Hwa
    • Korean Journal of Food Science and Technology
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    • v.33 no.6
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    • pp.669-675
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    • 2001
  • The objective of this study was to examine the effects of antioxidant vitamins on the formation of $AFB_{1}-DNA$ adduct and $AFB_{1}-inducing$ cellular oxidative damage. Intraperitoneal(i.p.)injections of 10 mg/kg vitamin C(VC) and 63.8 mg/kg vitamin E(VE) were repeatedly administrated 4 times with 2 days interval to 6 week old male ICR mice. After one hour of vitamin treatments, 0.4 mg/kg $AFB_1$ was injected in $AFB_1$ plus vitamin treated groups by same way. On the other hands, $AFB_1$ treated group was only injected with $AFB_1$ by the same method described above without vitamins. According to quantitative analysis of the $AFB_1$ in mice serum by indirect competitive ELISA, 12.28 and 18.78 ng/mL were detected in $AFB_1-treated$ groups, but 7.60 and 4.85 ng/mL in $AFB_1$ plus VC and VE treated groups, respectively. 23.78, 25.48 ng/mL of $AFB_1-DNA$ adduct were detected in mice liver of $AFB_1$treated groups, while 5.26, 7.81 ng/mL in $AFB_1$ plus VC and VE treated groups, respectively. Consequently, the differences in the concentrations of $AFB_1$ related materials between vitamin treated and non-treated groups were significant. Immunohistochemistry revealed brownish infiltration of $AFB_1$ around central vein and sinusoid in $AFB_1-treated$ group. This manifestation was distinctly reduced in $AFB_1$ plus VC and VE treated groups.

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A Study on the Effect of Herbal-acupuncture with Carthami Flos at Joksamni($ST_{36}$) on Collagen-induced Arthritis in Mice (족삼리(足三里) 홍화약침(紅花藥鍼)처치가 Collagen으로 유발한 생쥐의 관절염 모델에 미치는 영향)

  • Park, Ki-Hong;Lee, Hyun
    • Journal of Acupuncture Research
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    • v.24 no.6
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    • pp.45-61
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    • 2007
  • Objectives : The purpose of this study is to observe the effects of Carthami Flos herbal-acupuncture (CF-HA) at Joksamni($ST_{36}$) on arthritis in mice induced by Collagen II. Methods : The author performed several experimental items, including arthritis evaluation, change in weight, spleen size and stenosis rate, change in cytokine level, IgG, IgM and anti-collagen II, change of immunocyte count and histological change of the CIA mouse joint. Conclusions are as follows: Results : 1. In the CF-HA, the arthritis index and rate and the incidence of arthritis were decreased as the experiment proceeded. 2. In the CF-HA, spleen swell and stenosis, joint edema and change were decreased. 3. In the CF-HA, the level of $IL-1{\beta}$, IL-6, $TNF-{\alpha}$ and $IFN-{\gamma}$ in blood serum were significantly decreased. 4. In the CF-HA, the level of IgG, IgM and anti-collagen II were decreased. 5. In the CF- HA, $IFN-{\gamma}$, $IFN-{\gamma}/IL-4$, IL-10 of the culture fluid was decreased. 6. In the CF-HA, the cell rate of $CD3e^+$ and $CD45R^+$, $CD4^+$ and $CD8^+$, $CD4^+/CD25^+$ in spleen was similar to the cell rate of the normal group. 7. In the CF-HA, the cell rate of $CD4^+/CD25^+$, $CD45R^+/CD69^+$ in a lymph node was decreased as in the normal group. 8. In the CF-HA, the cell rate of $CD3^+/CD69^+$, $CD11b^+/Gr-1^+$ in joints was decreased as in the normal group. 9. In the CF-HA, the cartilage destruction and the inflammation cell growth in the H&E stain were decreased. The collagen fiber in the M&T stain were less destructed, therefore the result was similar to the normal group. Conclusions : These results suggest that CH-HA at $ST_{36}$ has an effect in controlling immune reaction and suppressing inflammation in rheumatoid arthritis therefore, the continuous flow of the following study is expected.

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Effect of Sinapis alba L. on expression of interferon-gamma and interleukin-4 production in anti-CD3/anti-CD28-stimulated CD4(+) T cells (CD4+ T cells에서 백개자가 IFN-$\gamma$와 IL-4 생성에 미치는 영향)

  • Park, Dae-Jung;Lee, Jang-Cheon;Lee, Young-Cheol
    • The Korea Journal of Herbology
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    • v.25 no.2
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    • pp.129-136
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    • 2010
  • Objective : Sinapis alba L. (SA) is a korean traditional herbal medicine that is usually used to prevent or treat inflammatory diseases, such as respiratory infection and rheumatoid arthritis. However, the effects of SA supplementation in vitro on serum antibody levels, splenocyte and peritoneal macrophage immune responses have not yet been determined. In this study, we examined the effect of SA on the production of Th1/Th2 cytokines. Methods : Splenocytes were isolated from naive C57BL/6 mice. Cells were enriched for CD4+ cell populations by first staining the cells with anti-CD4 (BD PharMingen, Calif, USA). CD4+ T cells were selected on a (CS) column, and the flow-through was collected as CD4+ T cells. Isolated cells were activated by overnight incubation on 24-well plates coated with $1{\mu}g/mL$ anti-CD3, $1{\mu}g/mL$ anti-CD28 and with SA ($100{\mu}g/mL$). Primary macrophages were collected from the peritoneal cavities of mice (8-week-old female C57BL/6). The peritoneal macrophages were washed and plated with RPMI-1640 overnight for the experiments. After 48-hours cultures, samples were centrifuged at 2000 rpm for 10 minutes, and the supernatants were stored at $-80^{\circ}C$. Mouse IL-4, IFN-$\gamma$ and TNF-$\alpha$ were quantified using ELISA kits (BioSource International, Camarillo, Calif, USA) according to the manufacturer's protocols. Results : SA at 100ug/ml decreased the generation of Th1 cytokine (IFN-$\gamma$) by 0.5-fold. However, SA has no effect on Th2 (IL-4) production. Conclusions : These results suggest that SA may play an important role in the control of T-cell-mediated autoimmunity by down-regulation of Th1 cytokine (especially IFN-$\gamma$, TNF-$\alpha$). These data may contribute to the design of new immunomodulating treatments for a group of autoimmune diseases.

Hypoglycemic Effect of Smallanthus sonchifolius (Yacon) Extracts on Animals with Streptozotocin-induced Diabetes (야콘 추출물이 Streptozotocin으로 유도된 당뇨 동물모델에서 혈당강하에 미치는 효과)

  • Oh, Hong-Geun;Kang, Young-Rye;Kim, Jung-Hoon;Moon, Dea-In;Seo, Min-Young;Park, Sang-Hoon;Choi, Kwang-Ho;Kim, Chang-Ryong;Kim, Sang-Hyun;Oh, Ji-Hyun;Kim, Sun-Young;Kim, Min-Gul;Chae, Soo-Wan;Kim, Ok-Jin;Lee, Hak-Yong
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.41 no.6
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    • pp.759-765
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    • 2012
  • This study was performed to investigate improvements in diabetes mellitus by extracts of yacon in streptozotocin (STZ)-induced diabetic rats. Fifty rats were divided into a normal group and four experimental groups. STZ (45 mg/kg) was injected intraperitoneally to induce type I diabetes in the four experimental groups. Yacon extracts were administered for 5 weeks. Forty-five ICR mice were also divided into one positive control group and four experimental groups for the oral glucose tolerance test (OGTT) after fed yacon extract. The control group did not eat any yacon extracts, while Group 1 (GI) was fed 125 mg/kg of yacon extracts, Group 2 (GII) was fed 250 mg/kg of yacon extracts, and Group 3 (GIII) was fed 500 mg/kg of yacon extracts. After treatment for 5 weeks, blood glucose in GIII group showed decreased tendency at the 5 week. In OGTT by glucose, the glucose level of yacon treatment group in diabetic rats was significantly decreased compared to the glucose level of the control group, but there was no difference in OGTT by maltose. In ICR mice, the glucose level of the experimental group in OGTT by maltose was significantly decreased compared to the control group. The area of the islets of Langerhans was increased by yacon treatment in a dose-dependent manner on diabetic rats. Insulin concentration of the GIII group was also decreased compared to the control group, while the concentration of fructosamine, total cholesterol, and triglycerides in serum showed no difference. OGTT by glucose or maltose in ICR mice or diabetic rats, area of the Islets of Langerhans, and insulin concentration improved. Yacon treatment may be a useful therapeutic and preventive strategy for diabetes mellitus.

Effect of Angelica keiskei Koidzumi Extract on Alcohol-Induced Hepatotoxicity In Vitro and In Vivo (In Vitro 및 In Vivo 알코올 유도 간 손상에 대한 신선초 추출물의 효과)

  • Lee, Jeong Yoon;An, Yeon Ju;Kim, Ji Won;Choi, Hyo-Kyoung;Lee, Yoo-Hyun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.45 no.10
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    • pp.1391-1397
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    • 2016
  • We investigated the hepatoprotective effects of Angelica keiskei Koidzumi extract (AK) in HepG2-overexpressing cytochrome P4502E1 (CYP2E1) and C57BL/6J mice. In HepG2 cells expressing CYP2E1, cell viability and catalase activity in the ethanol-AK co-treated group significantly increased compared to those in the ethanol-treated group. In the in vivo study with C57BL/6J mice, the AK-supplemented group with ethanol liquid diet showed significantly reduced hepatic markers, including serum aspartate aminotransferase, alanine aminotransferase, and ${\gamma}$-glutamyl transferase, compared to the ethanol group without AK supplementation. AK supplementation (20 mg/kg BW/d) also significantly attenuated reactive oxygen species generation and malondialdehyde level. Notably, a low dose of AK supplementation (20 mg/kg BW/d) suppressed expression of hepatic CYP2E1 and inhibited CYP2E1 enzyme activity. These data indicate that a low dose of AK supplementation could restrain alcohol-induced hepatic damage mediated by CYP2E1.

Safety Evaluation of 30 kGy Irradiated Chocolate Ice Cream (30 kGy 감마선 조사된 초코아이스크림의 안전성 평가)

  • Jeon, Young Eun;Yin, Xing Fu;Chung, Cha-Kwon;Kang, Il-Jun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.6
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    • pp.898-903
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    • 2013
  • This study was investigated the potential toxicity of gamma-irradiated chocolate ice cream for its future use in space. Chocolate ice cream was irradiated at a dose of 30 kGy at a temperature of $-20^{\circ}C$. For the animal study, AIN-93G was used as a control diet and irradiated and non-irradiated chocolate ice cream diets were administered to male and female ICR mice (ten mice per group) for three months. During the experimental period, the group fed irradiated chocolate ice cream did not show any changes in appearance, behavior, mortality, body weight, organ weight, or food consumption compared to the control. Also, all biochemical parameters, including hematology profiles, erythrocyte counts, and serum biochemical values were in normal ranges. In histopathological examinations of liver and kidney tissues, there were no significant differences between the control group and the group fed irradiated chocolate ice cream. These results indicate that chocolate ice cream irradiated at 30 kGy did not cause any toxic effects and could be applied for the development of safe and hygienic space food.