• Preventive Nutrition and Food Science
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• 제18권1호
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• pp.18-22
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• 2013

To investigate radical scavenging effects and protective activities of bitter melon (Momordica charantia) against oxidative stress, in vitro and a cellular system using LLC-$PK_1$ renal epithelial cells were used in this study. The butanol (BuOH) fraction of bitter melon scavenged 63.4% and 87.1% of 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals at concentrations of 250 and $500{\mu}g/mL$, respectively. In addition, the BuOH fraction of bitter melon effectively scavenged hydroxyl radicals (${\cdot}OH$). At all concentrations tested, the scavenging activity of the BuOH fraction was more potent than that of the positive control, ascorbic acid. Furthermore, under the LLC-$PK_1$ cellular model, the cells showed a decline in viability and an increase in lipid peroxidation through oxidative stress induced by pyrogallol, a generator of superoxide anion ($O_2{^-}$). However, the BuOH fraction of bitter melon significantly and dose-dependently inhibited cytotoxicity. In addition, 3-morpholinosydnonimine (SIN-1), a generator of peroxynitrite ($ONOO^-$) formed by simultaneous releases of nitric oxide and $O_2{^-}$, caused cytotoxicity in the LLC-$PK_1$ cells while the BuOH fraction of bitter melon ameliorated oxidative damage induced by $ONOO^-$. These results indicate that BuOH fraction of bitter melon has protective activities against oxidative damage induced by free radicals.

• Toxicological Research
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• 제28권3호
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• pp.165-172
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• 2012

Raphanus sativus (Cruciferaceae), commonly known as radish is widely available throughout the world. From antiquity it has been used in folk medicine as a natural drug against many toxicants. The present study was designed to evaluate the hepatoprotective activity of radish (Raphanus sativus) enzyme extract (REE) in vitro and in vivo test. The $IC_{50}$ values of REE in human liver derived HepG2 cells was over 5,000 ${\mu}g/ml$ in tested maximum concentration. The effect of REE to protect tacrine-induced cytotoxicity in HepG2 cells was evaluated by MTT assay. REE showed their hepatoprotective activities on tacrine-induced cytotoxicity and the $EC_{50}$ value was 1,250 ${\mu}g/ml$. Silymarin, an antihepatotoxic agent used as a positive control exhibited 59.7% hepatoprotective activity at 100 ${\mu}g/ml$. Moreover, we tested the effect of REE on carbon tetrachloride ($CCl_4$)-induced liver toxicity in rats. REE at dose of 50 and 100 mg/kg and silymarin at dose of 50 mg/kg were orally administered to $CCl_4$-treated rats. The results showed that REE and silymarin significantly reduced the elevated levels of serum enzyme markers induced by $CCl_4$. The biochemical data were supported by evaluation with liver histopathology. These findings suggest that REE, can significantly diminish hepatic damage by toxic agent such as tacrine or $CCl_4$.

• 생약학회지
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• 제30권3호
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• pp.255-260
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• 1999

In the previous report, E-kong-san, which is usually used for recovering health in traditional medicine, has been shown to decrease cisplatin induced nephrotoxicity in vivo and in vitro. The significant reduction of E-kong-san on the cisplatin induced nephrotoxicity led us to investigate whether the effect of this water extract was a result of triggering antioxidation. In monkey kidney Vero cells, E-kong-san at $5{\sim}10\;mg/ml$ was able to attenuate 2mM cisplatin-stimulated cell death by 46.8% and 31.8%, respectively. E-kong-san showed strong free radical scavengering activities on 1,1-diphenyl-2-picrylhydrazil (DPPH) radical and xanthine/xanthine oxidase (XOD) generated superoxide anion radical $(O_2^{-.})$. We further studied the effects of E-kong-san on lipid peroxidation in rat liver microsomes induced by enzymatic and nonenzymatic methods. Moreover, E-kong-san exhibited significant inhibition on both ascorbic $acid/Fe^{2+}$ and $ADP/NADPH/Fe^{3+}$ induced lipid peroxidation in rat liver microsomes. Based on these results, we suggest that E-kong-san protects the cisplatin induced cytotoxicity by its antioxidative mechanism.

• 한국미생물·생명공학회지
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• 제50권2호
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• pp.228-234
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• 2022

Helianthus tuberosus is perennial plant as Compositae family and is shown various physiological activities such as analgesic, antipyretic, anti-inflammatory, anti-fungal, anti-spasmodic, aperient, cholagogue, diuretic, spermatogenic, stomachic, and tonic effects. In this study, we investigated the antidiabetic and anti-inflammatory effects of pancake mixture powder (PM) supplemented with H. tuberosus (PMH) in rat skeletal muscle L6 cells and murine macrophage RAW 264.7 cells, respectively. PM and PMH inhibited in vitro α-glucosidase activity. Glucose consumption was increased by PM and PMH without cytotoxicity in rat myoblast L6 cells. Western blot analysis revealed that PM and PMH down-regulated glycogen synthase kinase (GSK)-3β activation in L6 cells. PM and PMH inhibited inflammatory mediator, nitric oxide (NO) production without cytotoxicity in LPS-induced RAW 264.7 cells. The anti-diabetic and anti-inflammatory effects of PMH was more stronger than those of PM. Anti-diabetic and anti-inflammatory effects of PMH would be due to functional characteristics of the supplemented H. tuberosus and the presence of garlic and onion used as ingredients of PM. Taken together, our results that addition of functional materials such as H. tuberosus in product has synergic effects and PMH is potential candidate for treatment of diabetes through inhibiting inflammation.

• 대한한방부인과학회지
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• 제22권1호
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• pp.41-52
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• 2009

Purpose: This study was designed to investigate the anti-tumor metastasis by immunomodulating effects of extracts of Prunellae Spica. Methods: Antimetastatic experiment was conducted in vivo by using colon 26-M3.1 carcinoma. And we observed cytotoxicity of Prunellae Spica on colon 26-M3.1 carcinoma, L5178Y-R lymphoma cell, hela cell and macrophage. To observe the immnomodulating effects of Prunellae Spica, we estimated IL-6, IL-10, IL-12, TNF-${\alpha}$ from peritoneal macrophages. And we evaluated the activation of NK cell by using anti-asialo-GM1 serum. Results: We found that the administration of Prunellae Spica extracts significantly inhibited tumor metastasis in vivo. In an in vitro cytotoxicity analysis, cell growth are closer to 100% in case of colon 26-M3.1 carcinoma, L5178Y-R lymphoma cell, hela cell at low concentration. In case of macrophage, cell proliferation is closer to 100% less than $62.5{\mu}g/m{\ell}$ of Prunellae Spica extracts. The level of cytokine such as IL-6, IL-10, IL-12 which stimulates Prunellae Spica extracts was increased in dose-dependent manner compared to the control group. TNF-${\alpha}$ is hardly secreted less than $250{\mu}g/m{\ell}$ The depletion of NK cells by anti-asialo GM1 serum partly abolished the inhibitory effect of Prunellae Spica on tumor metastasis. Conclusion: Prunellae Spica appears to have considerable activity on the anti-metastasis by activation the immune system such as macrophage and NK cell.

• 한방재활의학과학회지
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• 제24권3호
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• pp.11-27
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• 2014

Objectives This study was designed to evaluate the effects of Ephedra Sinica and Fibrosum Gypsum extract on obesity by using 3T3-L1 cells and high fat diet rats. Methods In vitro, Ephedra Sinica and Fibrosum Gypsum extract (50, 100, 200, $500{\mu}g/ml$) were added in 3T3-L1 cells. Cytotoxicity was measured by MTT assasy. Adipocyte differentiation was measured by Oil Red O staining, GPDH activity and $C/EBP{\alpha}$ protein expression. In vivo, Sprague-Dawley rats were divided into 5 groups : Normal diet group (Normal group), taken high fat diet and no treatment group (Control group), taken high fat diet and orally administered Ephedra Sinica and Fibrosum Gypsum extract daily (Group I: 50 mg/kg, Group II: 100 mg/kg, Group III : 200 mg/kg, oral). For 6 weeks of administration, body weight and the amount of food intake were measured once a week. After administration, blood analysis (AST, ALT, T-Bilirubin, BUN, RBC, Hb, HCT), serum lipid level (triglyceride, Total cholesterol, HDL, LDL), serum leptin level, epididymal adipose tissue weight and histological finding of liver were estimated. Results In vitro, The cytotoxicity was not significant. 3T3-L1 cell's differentiation was significantly decreased in Oil Red O staining, GPDH activity and $C/EBP{\alpha}$ protein expression. In vivo, Body weight and the amount of food intake, AST, ALT, Total cholesterol, TG, LDL, serum leptin, epididymal adipose tissue weight showed significant decrease in group I, group II and group III. There were no significant difference in T-bilirubin, BUN, RBC, Hb and HCT between all groups. HDL showed significant increase in group I, group II and group III. In histological finding of liver tissue, there were decreased adiposity and cytopathic effect in group I, group II and group III. Conclusions It is suggested that Ephedra Sinica and Fibrosum Gypsum extract can be used in the treatment of obesity.

• Journal of Preventive Medicine and Public Health
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• 제28권1호
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• pp.27-42
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• 1995

수은이 마우스의 세포성 및 체액성 면역반응에 미치는 영향을 밝히고자 실험을 통해 얻은 결과를 다음과 같이 요약하는 바이다. $10^{-2}mM$이상의 농도에서 수은은 시험관내의 마우스 비장세포에 대한 명백한 독성 작용(toxic effect)을 나타냈다. 수은에 의한 마우스 생존을(survival rate)은 0.8mg(per 100g body weight) 이하의 농도에서는 100%생존율을 나타냈으며, 0.9mg 이상의 농도에서는 5일 이내에 100% 치사율을 나타냈다. Subtoxic 농도의 수은 투여에 따른 Arthus 및 DTH반응의 결과는 $Arthus(T_3)$반응에 있어서 수은 투여군과 대조군 사이에 유의한 차이가 나타나지 않았던 반면, $T_{24},\;T_{48}$의 DTH반응은 수은 투여군이 대조군에 비하여 억제되었다. 수은 투여 마우스의 면양적혈구(SRBC)에 대한 항체반응을 측정한 결과 면양적혈구에 대한 총응집소가는 수은 투여 농도에 의존적으로 대조군의 값에 비해 억제되었다. 시험관 내에서 마이토젠에 대한 비장세포의 증식 반응은 수은 농도에 의존적으로 억제되었다. 수은 투여군의 IgE 생산은 농도 증가에 따라 $1.7\sim2.0$배의 수준으로 증가하였으며, $IgG_1$의 경우도 증가의 경향을 보였으나 대조군에 비해 큰 차는 아니었다. 이상의 결과를 살펴볼 때 수은 중독시 출현할 수 있는 면역반응의 변화로서, 초기에는 세포성 면역반응의 감소가 현저할 것으로 사료되나 중독이 진행됨에 따라 Ig농도의 상승을 제외한 체액성 면역반응의 전반적인 감소가 수반될 것으로 예측된다.

• 대한한방소아과학회지
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• 제21권3호
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• pp.33-55
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• 2007

Objectives In the present study, the author intended to investigate whether bojung-ikgitang-gamibang(BJGB) and seonbang-paedoktang(SBPT) significantly affect in vivo and in vitro mucin secretion from airway epithelial cells. Methods In vivo experiment, mice's mucin which is on a hypersecretion of airway mucin, mice's tracheal goblet cells in hyperplasia and mice's intraepithelial mucosubstances were exposed with SO2for3weeks. Effects of orally-administered BJGB and SBPT during 1 week on vivo mucin secretion and hyperplasia of tracheal goblet cells were assessed by using both enzyme-linked immunosorbent assay(ELISA) and staining goblet cells with alcian blue. In vitro experiment, confluent hamster tracheal surface epithelial(HTSE) cells were metabolically radiolabeled with 3H-glucosamine for 24hrs and chased for 30 min in the presence of each agent to figure out the effectiveness of 3H-mucin secretion. Total elution profiles of control spent media and treatment sample through Sepharose CL-4B column were analyzed. The effects of each agent on contractility of isolated tracheal smooth muscle and effects of each agent on MUC5AC gene expression in cultured HTSE cells were investigated. Also, possible cytotoxicities of each agent were assessed by measuring lactate dehydrogenase(LDH) release. Additionally, effects of BJGB and SBPT on both MUC5AC gene expression in cultured HTSE cells and TNF- or EGF-induced MUC5AC gene expression in human airway epithelial cells (NCI-H292) were investigated. Results (1) BJGB and SBPT inhibited hypersecretion of in vivo mucin. SBPT also inhibited the increase the number of goblet cells. However, BJGB did not affect the increase of number of goblet cells; (2) BJGB significantly increased mucin secretion from cultured HTSE cells, without significant cytotoxicity, and chiefly affected the 'mucin' secretion; (3) SBPT did not affect mucin secretion from cultured HTSE cells without significant cytotoxicity, and also did not affect the secretion of the other releseable glycoproteins; (4) BJGB and SBPT did not affect Ach-induced contraction of isolated tracheal smooth muscle; (5) SBPT significantly inhibit the expression levels of MUC5AC gene and BJGB significantly increased the expression levels of MUC5AC gene in both HTSE cells and NCI-H292 cells. Conclusions BJGB and SBPT can not only affect the secretion of mucin but also affect the expression of mucin gene. The author suggests that the effects BJGB and SBPT with their components should be further investigated and it is highly desirable to find from oriental medical prescriptions, novel agents which might regulate hypersecretion of mucin from airway epithelial cells.

• Asian Pacific Journal of Cancer Prevention
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• 제15권22호
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• pp.9667-9672
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• 2014

Background: Pancreatic adenocarcinoma is a malignant gastrointestinal cancer with significant morbidity and mortality. Despite severe side effects of chemotherapy, the use of immunotherapy combined with chemotherapy has emerged as a common clinical treatment. In this study, we investigated the efficacy of the combined doxorubicin and interferon-${\alpha}$ (IFN-${\alpha}$) therapy on murine pancreatic cancer Panc02 cells in vitro and in vivo and underlying mechanisms. Materials and Methods: A Panc02-bearing mouse model was established to determine whether doxorubicin and interferon-${\alpha}$ (IFN-${\alpha}$) could effectively inhibit tumor growth in vivo. Cytotoxicity of natural killer (NK) cells and cytotoxic T lymphocytes (CTLs) was evaluated using a standard LDH release assay. To evaluate the relevance of NK cells and CD8 T cells to the combination therapy-mediated anti-tumor effects, they were depleted in tumor-bearing mice by injecting anti-asialo-GM-1 antibodies or anti-CD8 antibodies, respectively. Finally, the influence of doxorubicin+interferon-${\alpha}$ (IFN-${\alpha}$) on the ligands of NK and T cells was assessed by flow cytometry. Results: The combination therapy group demonstrated a significant inhibition of growth of Panc02 in vivo, resulting from activated cytotoxicity of NK cells and CTLs. Depleting CD8 T cells or NK cells reduced the anticancer effects mediated by immunochemotherapy. Furthermore, the doxorubicin+IFN-a treatment increased the expression of major histocompatibility complex class I (MHC I) and NKG2D ligands on Panc02 cells, suggesting that the combined therapy may be a potential strategy for enhancing immunogenicity of tumors. All these data indicate that the combination therapy using doxorubicin and interferon-${\alpha}$ (IFN-${\alpha}$) may be a potential strategy for treating pancreatic adenocarcinoma.

• 대한한방종양학회지
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• 제5권1호
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• pp.61-75
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• 1999

To examine the effect of Xuefuzhuyutang on the metastasis of cancer, the following experiments were carried out. Before the main experiments, the cytotoxicity was measured by putting Xuefuzhuyutant sample in HT1080. Then zymography was made to examine the change of gelatinolytic activity. Western blotting was carried out to examine the changes of Fos, Jun, Ets, Erk, md JNK. In vitro invasion assay with transwells coated by collagen and matrigel was carried out. From the above results the following conclusions were obtained. 1. The experimental result about cytotoxicity of Xuefuzhuyutang agaitst HT1080 was a below. The stained cell count after being treated by by Xuefuzhuyutang sample $400{\mu}g/ml$ for 24 hours was 0.9% of total cells, and the stained cell count by Xuefuzhuyutang sample $100{\mu}g/ml$ was 1.5% of total cells. Both were near the level of control group which showed 0.6% stained. 2. The result of collagenase assay was as below. In Xuefuzhuyutang sample $400{\mu}g/ml$, MMP2 was reduced as compared with TPA control group, and the band of MMP-9 induced by TPA disapappeared. In Xuefuzhuyutang samle $800{\mu}g/ml$ both bands of MMP-2 and MMP-9 disapeared. 3. The results of western blots for Jun, Fos, Ets, Erk, JNK were a below. In Xuefuzhuyutang sample $200{\mu}g/ml$, Ets was reduced, and Jun, Fos were increased. 4 The result of invasion assay was as below. The number of cells which migrated across transwell membrane in Xuefuzhuyutang-treated group was less than that of control(+TPA) group. From the above results, it was concluded that Xuefuzhuyutang might inhibit the activity of collagenase not by the MMP-2, MMP-9 promoter but by the other way.