• Title/Summary/Keyword: Western Blot

Search Result 2,774, Processing Time 0.028 seconds

Effects of Doinsenggitang on Melanin Synthesis and Gene Expression Inhibition in B16F10 Melanoma Cells (도인승기탕의 B16F10 세포주에서의 멜라닌 생성 및 유전자 발현 억제 효과)

  • Hwang, Ju-Young;Kim, Dong-Hee;Kim, Hui-Jung;Hwang, Eun-Young;Park, Tae-Soon;Lee, Jin-Young;Son, Jun-Ho
    • Journal of Life Science
    • /
    • v.22 no.3
    • /
    • pp.318-323
    • /
    • 2012
  • This study involved observation of the inhibitory effect of 70% EtOH and water extracts from Doinseunggitang on melanin synthesis, tyrosinase activity, and western blotting using B16F10 melanoma cells. Doinseunggitang extracts inhibited melanin synthesis and tyrosinase activity in a dependent manner. As a result, it was found that Doinseunggitang 70% EtOH extracts inhibit melanin synthesis and tyrosinase activity, respectively, by 40% and 51%. In addition, western blotting analysis showed that 70% EtOH extracts inhibited tyrosinase, MITF, TRP-1, and TRP-2 expression. These results show that 70% ethanol extracts of Doinseunggitang could be developed as a skin whitening material in cosmetics.

Expression of Dopamine D2 Receptor in Response to Apomorphine Treatment in the Striatum of the Rat with Experimentally Induced Parkinsonism (파킨슨병 모형 흰쥐의 줄무늬체에서 Apomorphine 투여 방법에 따른 도파민 D2 수용체의 발현)

  • Choi, Seung Jin;Sung, Jae Hoon;Son, Byung Chul;Park, Choon Keun;Kwon, Sung Oh;Kim, Moon Chan;Lee, Sang Won
    • Journal of Korean Neurosurgical Society
    • /
    • v.29 no.7
    • /
    • pp.868-876
    • /
    • 2000
  • Objective : Parkinsonian rat models have generally been characterized by unilateral destruction of both the nigrostriatal pathway and the mesolimbic pathway using the neurotoxin 6-hydroxydopamine. The induction of contraversive turning by apomorphine in these models is thought to reflect the stimulation of supersensitive dopamine D2 receptor or receptor-mediated mechanisms in denervated neostriatum. The present study was undertaken to investigate the expression of dopamine D2 receptor in denervated striatum according to modalities of apomorphine(dopamine agonist) treatment after creating a hemiparkinsonian rat model in which there is 6-hydroxydopamine induced destruction of the unilateral dopaminergic nigrostriatal pathway. Methods : After making complete lesion in left side substantia nigra pars compacta(SNpc) by stereotactic injection of 6-hydroxydopamine into medial and lateral areas of SNpc, and confirming successful animal model by apomorphine induced contraversive turning behavior without recovery and complete destruction of ipsilateral SNpc with tyrosine hydroxylase immunostaining in 7th day after operation, 15 rats of parkinsonian model were studied with or without administration of apomorphine at varying doses and durations. According to the modalities of apomorphine treatment for 4 days, these rats were divided into 3 groups, as not-treated group, intermittently treated group and constantly treated group. For investigating the extent of the expression of dopamine D2 receptor in denervated striatum, immunohistochemical staining by dopamine D2 receptor antibody and Western blot were performed. Results : In the D2 receptor antibody immunohistochemical staining, the mean number of positive stained neurons was highest in not-treated group($20.5{\pm}1.14$) of 3 groups. In constantly treated group, the mean number of positive stained neurons was less($3.9{\pm}1.79$) than intermittently treated group(p<0.05). The Western blotting with the D2 receptor antibody revealed that expression of receptors was also highest in not-treated group and less in constantiy treated group than intermittently treated group. Conclusion : Dopamine D2 receptors in denervated striatum of parkinsonian rat models, which were not treated with apomorphine, revealed to be most highly expressed. And, according to doses and durations of apomorphine administration, desensitization of the receptor was more apt to develop with constant treatment than intermittent treatment. In clinical setting, the authors believe that, in long-term treated parkinsonian patients, desensitization of dopamine receptors due to chronic dopaminergic stimulation seems to be partially related to mechanisms of drug tolerance.

  • PDF

Verification of Anti-inflammatory Activities of the Ethanol Extracts of Glechoma hederacea var. longituba in RAW 264.7 Cells (RAW 264.7 세포에서 긴병꽃풀 에탄올 추출물의 항염증 활성 검증)

  • Lee, Jin-Young;Yoo, Dan-Hee;Jeong, Yong-Seong;Joo, Sung-Hyun;Chae, Jung-Woo
    • Journal of Life Science
    • /
    • v.28 no.4
    • /
    • pp.429-434
    • /
    • 2018
  • In this study, we investigated the potential of Glechoma hederacea var. longituba 70% ethanol extract as a natural functional material by examining the anti-inflammatory effect of it. Macrophages results in (Raw 264.7) induced by lipopolysaccharide (LPS). Confirming the viability of the macrophages Glechoma hederacea var. longituba 70% ethanol extract showed a 95.8% survival rate at $1,000{\mu}g/ml$ concentration. Anti-inflammatory activity was examined the inhibitory tests on the production of LPS included nitric oxide (NO) in RAW 264.7 cells by Griess assay. The result showed that NO production deterrent effect of 37.4% at a concentration of $1,000{\mu}g/ml$. The deterrent effect of GG 70% ethanol extract on protein expression of inducible NOS (iNOS) and Cyclooxygenase-2 (COX-2) was measured by Western blotting using the concentrations 50, 100 and $500{\mu}g/ml$, with ${\beta}$-actin used as the positive control. The inhibitory effect of iNOS and COX-2 mRNA expression was measured by reverse transcription-polymerase chain reaction (RT-PCR) using 50, 100 and $500{\mu}g/ml$ concentration of GG 70% ethanol extract, with GAPDH used as the positive control. In experiments using Western blot and RT-PCR when compared with the control group vitamin C it was confirmed that the 70% ethanol extract from GG suppressed. When compiling the results of this study, we confirmed the possibility of GG 70% ethanol extract as an anti-inflammatory material.

Immune Response of Bacterial Proteins of Staphylococcus intermedius from Canine Atopic Dermatitis (개의 아토피성 피부염에서 분리한 Staphylococcus intermedius 균의 세균단백질의 면역반응)

  • Park, Hee-myung
    • Journal of Veterinary Clinics
    • /
    • v.21 no.1
    • /
    • pp.20-22
    • /
    • 2004
  • Bacterial infection of canine atopic dermatitis is largely caused by Staphylococcus intermedius and may be a superficial or deep pyoderma. The Purpose of this study was to identify the major proteins of S. intermedius cell surface components in humoral immune response of atopic dermatitis dog. Sera samples were obtained from dogs with atopic dermatitis and superficial pyoderma referred to the Veterinary Medical Teaching Hospital, Konkuk University. An isolate of S. intermedius from a clinical case of canine atopic dermatitis was cultured in brain heart infusion broth overnight at $37^{\circ}C$ in aerobic conditions on an orbital shaker. Following culture, Staphylococci were harvested by centrifugation, washed in PBS, and resuspended in PBS containing lysostaphin. The soluble components were separated by centrifugation and were collected. The soluble extract of S. intermedius was separated by sodium dodecylsulphate polyacrylamide gel electrophoresis (SDS-PAGE). The proteins were electrophoretically transferred onto nitrocellulose membrane. Western blotting for the specificity of serum IgG antistaphylococcal antibody was performed with anti-dog-IgG and sera obtained from an atopic dermatitis case and a normal dog. The molecular masses of four major proteins of S. intermedius recognized by serum obtained from an atopic dermatitis case were 18, 31, 75, and 110 kDa as determined by Western blot analysis. The present study indicates that most dogs of S. intermedius infection with atopic dermatitis could have a significant humoral immune response to bacterial proteins of the causative organism.

Anti-inflammatory Activities Verification of Vaccinum oldhami Fruit Ethanol Extracts on RAW 264.7 (RAW 264.7을 이용한 정금나무 열매(Vaccinum oldhami fruit)의 항염증 효과)

  • Lee, Jin-Young;Joo, Da-Hye;Yoo, Dan-Hee;Chae, Jung-Woo
    • Journal of Life Science
    • /
    • v.27 no.4
    • /
    • pp.417-422
    • /
    • 2017
  • The purpose of this study was to investigate the role of the Vaccinum oldhami fruit extract as a cosmetic additive. As a result of having macrophage (RAW 264.7) measured a cell toxicity effects of 70% ethanol extract from Vaccinum oldhami fruit, it shown 118% with toxicity at $500{\mu}g/ml$ concentration. In nitric oxide synthesis inhibition effect, 70% ethanol extracts from Vaccinum oldhami fruit shown 47.3% at $1,000{\mu}g/ml$ concentration. The iNOS, COX-2 protein expression inhibitory effect by western blot of 70% ethanol extract from Vaccinum oldhami fruit was decreased by 36.13%, 29.61% at $500{\mu}g/ml$ concentration. And iNOS, COX-2 mRNA expression inhibitory effect by reverse-transcription-PCR of 70% ethanol extract from Vaccinum oldhami fruit was decreased by 62.25%, 90.07% at $500{\mu}g/ml$ concentration. All these finding that extract from Vaccinum oldhami fruit could prove that their have effects anti-inflammatory efficacy. And extract from Vaccinum oldhami fruit has potential as a cosmetic ingredients.

Measurement of Mucin Amounts Using RTO3 in Patients with Chronic Airway Disease (만성 기도질환에서 흰쥐 뮤신에 대한 단 세포군 항체(RTO3)를 이용한 뮤신 측정에 관한 연구)

  • Kim, Do-Jin;Kim, Ki-Up;NamGung, Eun-Kyang;Uh, Soo-Taek;Kim, Young-Hoon;Shin, Chan-Young;Ko, Kwang-Ho;Park, Choon-Sik
    • Tuberculosis and Respiratory Diseases
    • /
    • v.47 no.6
    • /
    • pp.786-796
    • /
    • 1999
  • Background: It has been anticipated that the amount and composition of mucin are changed in patients with chronic airway diseases. We evaluated whether RTO3(mAb against rat tracheal mucins) could quantify the amount of mucin from the airway in the patients with chronic airway diseases. Methods and results; 1) RTO3 was bound to high molecular weight of mucin based on Western blot in sputum and BALF from patients with chronic airway diseases. 2) The goblet cells and submucosal glands in main bronchus from human were observed by PAS stain. And immunohistochemical stain with RTO3 showed immunoreactivity on some goblet cells. 3) The amount of mucin was more increased in patients with chronic airway diseases compared to those in normal subjects. 4) In the exacerbation of asthmatics, mucin amounts were more increased than stable asthmatics. Conclusion: We suggested that secreted mucin in chronic airway diseases can be quantified by ELISA with RTO3.

  • PDF

Protective effect of Artemisiae Capillaris Herba water extract on liver injury induced by thioacetamide (인진호 열수 추출물이 thioacetamide에 의해 유발된 간손상에 미치는 간보호 효과)

  • Kim, Min Ju;Lee, Jin A;Shin, Mi-Rae;Park, Hae-Jin;Roh, Seong-Soo
    • Journal of Nutrition and Health
    • /
    • v.54 no.4
    • /
    • pp.412-421
    • /
    • 2021
  • Purpose: Thioacetamide (TAA) produces reactive oxygen species (ROS) in the liver, and the generated ROS induces liver injury through inflammatory reactions. The current study was undertaken to investigate the hepatoprotective effect of Artemisiae Capillaris Herba water extract (AC), imparted via its antioxidant activity, in an animal model of TAA-induced liver injury. Methods: Animal experiments were conducted in 5 groups: normal, control (TAA 200 mg/kg), SM (TAA 200 mg/kg + silymarin 100 mg/kg), ACL (TAA 200 mg/kg + AC 100 mg/kg), ACH (TAA 200 mg/kg + AC 200mg/kg). TAA (intraperitoneal) and treatment compounds (per oral) were administered for 3 days. Serum levels of ammonia concentration and myeloperoxidase (MPO) activity were subsequently measured. Liver tissues were subjected to western blot analysis for measuring the oxidative stress (NADPH oxidase), anti-oxidative activity (Nrf2, heme oxygenase-1 [HO-1], superoxide dismutase [SOD], catalase, and GPx-1/2), tissue inhibitors of metalloproteinase (TIMP)-1, and matrix metalloproteinases (MMPs) protein expressions. Results: Serum ammonia levels and MPO activity were significantly increased in the TAA-induced control group, whereas groups administered AC treatment showed markedly reduced levels. Western blot analysis revealed significantly increased NOX2 and p22phox expressions, (oxidative stress-related factors) in the TAA-induced control group. These levels were determined to be significantly decreased after AC exposure. Moreover, antioxidant-related factors including Nrf2, HO-1, SOD, catalase, and GPx-1/2 were significantly decreased in the control group and increased in the AC treated groups. In addition, MMP expressions were significantly suppressed in the AC treatment group due to increased levels of TIMP-1. Conclusion: Taken together, these data indicate that exposure to AC reduces the oxidative stress by inhibiting the expression of NADPH oxidase (NOX2 and p22phox) through the Nrf2 signaling pathway. We therefore propose the potential of AC for the prevention and treatment of TAA-induced liver injury.

A Sphingosine Kinase-1 Inhibitor, SKI-II, Induces Growth Inhibition and Apoptosis in Human Gastric Cancer Cells

  • Li, Pei-Hua;Wu, Jin-Xia;Zheng, Jun-Nian;Pei, Dong-Sheng
    • Asian Pacific Journal of Cancer Prevention
    • /
    • v.15 no.23
    • /
    • pp.10381-10385
    • /
    • 2015
  • SKI-II has been reported as an inhibitor of sphingosine kinase 1 and has been extensively used to prove the involvement of sphingosine kinase and sphingosine-1-phosphate (Sphk1) in cellular processes. In the current study, we investigated the effects of SKI-II and its potential mechanisms in human gastric cancer SGC7901 cells. After treatment with SKI-II, cell growth, cell cycle distribution, apoptosis, expression of Sphk1, NF-${\kappa}B$, Bcl-2, Bax and p27 were assessed by MTT assay, flow cytometry, electron microscopy, immunocytochemistry and Western-blot assay, respectively. Our results showed that SKI-II markedly inhibited SGC7901 cell survival in a dose-dependent manner, reduced cell proliferation with accumulation of cells in the G0/G1 phase and induced apoptosis in the tumor cells. Furthermore, Western blotting and immunocytochemistry showed that the expression of p27 and Bax was increased significantly, but the expression of NF-${\kappa}B$, Bcl-2 and Sphk1 decreased by different degrees. These results indicate that SKI-II induced cell growth arrest and apoptosis. The increased apoptotic sensitivity of SGC7901 was correlated with NF-${\kappa}B$ or Bcl-2/Bax activation.

Momordica cochinchinensis Seed Extracts Suppress Migration and Invasion of Human Breast Cancer ZR-75-30 Cells Via Down-regulating MMP-2 and MMP-9

  • Zheng, Lei;Zhang, Yan-Min;Zhan, Ying-Zhuan;Liu, Chang-Xiao
    • Asian Pacific Journal of Cancer Prevention
    • /
    • v.15 no.3
    • /
    • pp.1105-1110
    • /
    • 2014
  • Objective: Metastases and invasion are the main reasons for oncotherapy failure. Momordica cochinchinensis (Mu Bie Zi in Chinese) had been used for a variety of purposes, and shown anti-cancer action. In this article, we focused on effects on regulation of breast cancer cell ZR-75-30 metastases and invasion by extracts of Momordica cochinchinensis seeds (ESMCs). Methods: Effect of ESMCs on ZR-75-30 human breast cancer cells proliferation were evaluated by MTT assay and on invasion and migration by wound-healing and matrigel invasion chamber assays. Expression and protease activity of two matrix metalloproteinases (MMPs), MMP-2 and MMP-9, were analyzed by Western blotting and gelatin zymography, respectively. Results: ESMC revealed strong growth inhibitory effects on ZR-75-30 cells, and effectively inhibited ZR-75-30 cell invasion in a dose-dependent manner. Western blot and gelatin zymography analysis showed that ESMC significantly inhibited the expression and secretion of MMP-2 and MMP-9 in ZR-75-30 cells. Conclusions: ESMC has the potential to suppress the migration and invasion of ZR-75-30 cancer cells, and it might prove to of interest in the development of novel inhibitors for breast cancer.

Terpinen-4-ol Induces Autophagic and Apoptotic Cell Death in Human Leukemic HL-60 Cells

  • Banjerdpongchai, Ratana;Khaw-on, Patompong
    • Asian Pacific Journal of Cancer Prevention
    • /
    • v.14 no.12
    • /
    • pp.7537-7542
    • /
    • 2013
  • Background: Terpinen-4-ol, a monoterpene, is found as the main component of essential oil extracts from many plants. In this study apoptotic and autophagic types of cell death induced by terpinen-4-ol and associated mechanisms were investigated in human leukemic HL-60 cells. Materials and Methods: The cytotoxicity of human leukemic U937 and HL-60 cells was determined by MTT assay. Cytochrome c release, expression of Bax, Bcl-2, Bcl-xl and cleaved Bid were determined by Western blotting. Cell morphology was examined under a transmission electron microscope. LC3-I/II, ATG5 and Beclin-1 levels were detected by immunoblotting. Results: Terpinen-4-ol exhibited cytotoxicity to human leukemic HL-60 but not U937 cells. The apoptotic response to terpinen-4-ol in HL-60 cells was due to induction of cytochrome c release from mitochondria and cleavage of Bid protein after the stimulation of caspase-8. There was a slightly decrease of Bcl-xl protein level. The characteristic cell morphology of autophagic cell death was demonstrated with multiple autophagosomes in the cytoplasm. At the molecular level, the results from Western blot analysis showed that terpinen-4-ol significantly induced accumulation of LC3-I/II, ATG5 and Beclin-1, regulatory proteins required for autophagy in mammalian cells. Conclusions: Terpinen-4-ol induced-human leukemic HL-60 cell death was via both autophagy and apoptosis.