• Title/Summary/Keyword: in vitro suppression

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Ghrelin Attenuates Dexamethasone-induced T-cell Apoptosis by Suppression of the Glucocorticoid Receptor (덱사메타손에 의해 유발된 흉선 T세포사멸에 대한 그렐린의 세포사멸억제효과)

  • Lee, Jun Ho
    • Journal of Life Science
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    • v.24 no.12
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    • pp.1356-1363
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    • 2014
  • Ghrelin is a 28 amino acid orexigenic peptide hormone that is secreted predominantly by tX/A cells in the stomach, and it plays a major role in energy homeostasis. Activated ghrelin has an n-octanoyl group covalently linked to the hydroxyl group of the Ser3 residue, which is critical for its binding to the G-protein coupled growth hormone secretagogue receptor-1a (GHS-R1a). According to recent reports, both ghrelin and its receptor, GHS-R1a, are expressed by a variety of immune cells, including T- and B-lymphocytes, monocytes, and dendritic cells, and ghrelin stimulation of leukocytes provides a potent immunomodulatory signal controlling systemic and age-associated inflammation and thymic involution. Here, we report that ghrelin protected murine thymocytes from dexamethasone (DEX)-induced cell death both in vivo and in vitro. Subsequently, we explored the molecular mechanisms of the antiapoptotic effect of ghrelin. According to our experiments, ghrelin inhibited the expression of proapoptotic proteins via the regulation of glucocorticoid receptor (GR) phosphorylation. As a result, ghrelin inhibited the proapoptotic activation of proteins, such as Caspase-3, PARP, and Bim. These data suggest that ghrelin, through GHS-R, inhibits the pathway to apoptosis by regulation of the proapoptotic protein activation signal pathway. They provide evidence that blocking apoptosis is an essential function of ghrelin during the development of thymocytes.

Effects of Egg Yolk Antibodies Produced in Response to Different Antigenic Fractions of E. coli O157:H7 on E. coli Suppression

  • Chae, H.S.;Singh, N.K.;Ahn, C.N.;Yoo, Y.M.;Jeong, S.G.;Ham, J.S.;Kim, D.W.
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.11
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    • pp.1665-1670
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    • 2006
  • The objective of this research was to provide the characterization and method for producing anti-E. coli O157:H7 antibodies in egg-laying hens and to determine if the antibody can restrain the proliferation of E. coli O157:H7 in-vitro. Selected antigenic fractions (whole cell, outer membrane protein and lipopolysaccharide (LPS)) from E. coli O157:H7 were injected to hens in order to produce anti-E. coli O157:H7 antibodies. The immune response and the egg yolk antibodies of laying hens against the whole cell, outer membrane protein and LPS antigens were monitored by ELISA. The level of antibodies against whole cell antigen monitored through ELISA sharply increased after the initial immunization, and it was found to be maximum on day 49 however, the level was maintained up to day 70. Antibodies (5 mg/ml) directed against the whole cell inhibited E. coli proliferation 10-13 times more than outer membrane protein or LPS. The antibody response against the whole cell antigens appeared to have higher activity in restraining the proliferation of E. coli O157:H7 than antibody against outer membrane protein or LPS. Results reflected that increasing the IgY's in the egg yolk could prevent greater economic losses due to human and animal health from pathogenic bacteria i.e. E. coli O157:H7.

Studies on the Development of Antihyperlipidemic Drugs from Oriental Herbal Medicines(I) -Antihyperlipidemic Activities of Oriental Herbal Medicines- (한방약물로부터 항고지혈증 치료약물개발(I) -수종 한약재의 항고지혈증 활성검색-)

  • Jung, Eun-Ah;Kim, Dong-Hyun;Lee, Sang-In;Kim, Nam-Jae
    • Korean Journal of Pharmacognosy
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    • v.30 no.4
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    • pp.368-376
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    • 1999
  • Twenty-four Oriental herbal medicines including Platycodi Radix and Scutellariae Radix, etc., which have been used for the cure of hyperlipidemia, coronary heart, disease were evaluated for the 3-hydroxy-3-methylglutaryl coenzyme A(HMG-CoA) reductase inhibition and antioxidant effect on a free radical, 1,1-diphenyl-2-picrylhydrazyl(DPPH) in vitro, and antihyperlipidemic effect on hyperlipidemic rats induced by Triton WR-1339 in vivo. 80% MeOH extract of eight herbs including Scutellariae Radix, Coptidis Rhizoma, Paeoniae Radix Rubra, Moutan Cortex Radicis, Angelicae Gigantis Radix, Bambusae Caulis in Taeniam, Cinnamomi Ramulus and Crataegi Fructus inhibited the HMG-CoA reductase activities and exhibited a radical-trapping action on a stable free radical, DPPH. On Triton WR-1339-induced hyperlipidemic rats, four herbs including Scutellariae Radix, Angelicae Gigantis Radix, Bambusae Caulis in Taeniam and Cinnamomi Ramulus showed respectively the significant suppression of serum total cholesterol, triglyceride, phospholipid and LDL-cholesterol levels and serum transaminase(ALT and AST) activities. From these results, it is suggested that each 80% MeOH extract of Scutellariae Radix, Angelicae Gigantis Radix, Bambusae Caulis in Taeniam and Cinnamomi Ramulus have effective antihyperlipidemic action against hyperlipidemia.

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Effect of Acibenzolar-S-methyl and Rahnella aquatilis (Ra39) on Chitinase and β-1, 3-glucanase Activities and Disease Resistance of Apple Plants

  • Abo-Elyousr, A.M. Kamal;Sallam, M.A.A.;Hassan, M.H.A.;Zeller, W.
    • The Plant Pathology Journal
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    • v.26 no.1
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    • pp.63-69
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    • 2010
  • The effect of Acibenzolar-S-methyl (ASM) and Rahnella aquatilis Ra39 against apple fire blight disease caused by Erwinia amylovora were tested as a possible alternative to streptomycin. In vitro studies, no inhibition effect against the pathogen was found when ASM was tested. Under greenhouse conditions, application of R. aquatilis Ra39 with the highly susceptible M26 rootstock resulted in a marked disease suppression. Application of ASM and strain Ra39 caused a high decrease of the disease, 82% and 58% respectively; this was correlated with a reduction of the growth of the pathogen within host plants up to 64% and 49.5% respectively. Further studies in the field under artificial infection condition during full bloom revealed that application of ASM and R. aquatilis Ra39 with Gala variety resulted in a control effect up to 21 and 29% respectively. In physiological studies, enhanced activities of PR-proteins (chitinase and $\beta$-1, 3-glucanase) were detected, which are well known as biochemical markers for systemic acquired resistance. Application of ASM to apple shoots caused the highest chitinase activity followed by strain Ra39. The enzyme activity was increased after 2, 4 and 6 days from application. In addition, ASM-treatment caused the higher $\beta$-1, 3-glucanase activity than strain Ra39. Maximum enzyme activity was recorded after 6 days from application and then decreased after 8 and 10 days from application.

The Effect of L-carnitine and Acetylcarnitine on Sperm Parameters in vitro (생체외 L-carnitine과 Acetylcarnitine의 정자지표 개선 효과)

  • Lee, Wan;Park, Nam-Cheol
    • Clinical and Experimental Reproductive Medicine
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    • v.29 no.2
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    • pp.149-157
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    • 2002
  • Objectives: To assess the scavenging effect of carnitine derivatives on oxidative damage to sperm during sperm processing, cryopreservation and thawing. Materials and Methods: Fresh semen samples from 20 normal healthy volunteers were collected by masturbation after at least 48 hours abstinence. After liquefaction of semen samples at room temperature, the specimens were diluted with sperm wash media (Ham's F-10, Life technologics) to a uniform density of $20{\times}10^6/ml$. L-carnitine or acetylcarnitine were added with various concentration of $0{\mu}M$, $10{\mu}M$, $30{\mu}M$ in semen sample or cryoprotectant. All specimens were cryopreservated at $-196^{circ}C$ $LN_2$ for 3 days. Sperm motility, vitality, fertilizing capacity, reactive oxygen species formation and the level of lipid peroxidation were analyzed by computer assisted semen analyzer, eosin-nigrosin stain, hypoosmotic swelling test, chemiluminescence and thiobarbituric acid method, respectively, during sperm processing, cryopreservation and thawing. Results: The sperm motility was only increased in proportion to the concentration of acetylcarnitine with no statistical significance (p>0.05). The sperm vitality was also significantly improved in proportion to the concentration of acetylcarnitine with statistical significance (p<0.05). The sperm fertilizing capacity was significantly increased in proportion to the concentration of L-carnitine and acetylcarnitine and reactive oxygen species generation and lipid peroxidation were significantly decreased with same fashion (p<0.05). On comparison of effects between L-carnitine and acetylcarnitine, acetylcarnitine was superior to L-carnitine on the improvement of sperm motility and vitality as well as the suppression of reactive oxygen species generation and lipid peroxidation. Conclusions: These results suggest that carnitine derivatives have a scavenging effect against oxidative damages during sperm processing, cryopreservation and thawing. Therefore, carnitine derivatives may be useful as an oral antioxidant in patients with male infertility due to increased ROS generation.

Protective Effects and Mechanisms of Salvia Miltiorrhizae Radix Extract on Endotoxin Shock (패혈증 쇼크에서 단삼 추출물의 억제 효과와 그 기전에 관한 연구)

  • Shin, Sang-Ho;Yun, Jeong-Min;Ko, Woo-Shin;Yoon, Hwa-Jung
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.29 no.3
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    • pp.14-26
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    • 2016
  • Objectives : This study was performed to investigate the protective effects and mechanisms of Salvia miltiorrhizae Radix extract (SME) on endotoxin shock.Methods : We used two models; LPS-induced sepsis model for in vivo model, and murine peritoneal macrophages responses for in vitro. SME was administrated orally to mice. After 1 hr, LPS was injected intraperitoneally. Survival rate was checked each time per 12 hr for 5 days. Mice were sacrificed 3 hr after LPS injection, then blood samples and organs were harvested. Cytokines secretion was measured by ELISA. Organs tissues were observed with microscope. Murine peritoneal macrophages were cultured for 1 hr either in a medium alone or in a medium that contained SME, as indicated. Then, the cells were treated with LPS for 24 hr. mRNA levels of cytokines were measured by real-time RT-PCR. Cytokine levels in the supernatants were measured by ELISA. The amount of nitrite was measured by using the Griess method to evaluate NO production. The cell lysates were analysed by Western blotting using antibodies for iNOS and β-actin was used as an internal control to monitor equal protein loading.Results : SME improverd the survival rate of mice model. SME inhibited the secretion of inflammatory cytokines and organs damages on Endotoxin Shock model. SME suppressed cytokine expression, cytokine secretion,NO production, iNOS expression in LPS-induced murine peritoneal macrophages.Conclusions : The results suggest that SME has protective effects on endotoxin shock through suppression of inflammatory cytokines, organ damages, NO production and so on.

Bidirectional Regulation of Manganese Superoxide Dismutase (MnSOD) on the Radiosensitivity of Esophageal Cancer Cells

  • Sun, Guo-Gui;Hu, Wan-Ning;Wang, Ya-Di;Yang, Cong-Rong;Lu, Yi-Fang
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.7
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    • pp.3015-3023
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    • 2012
  • The mitochondrial antioxidant protein manganese superoxide dismutase (MnSOD) may represent a new type of tumor suppressor protein. Overexpression of the cDNA of this gene by plasmid or recombinant lentiviral transfection in various types of cancer leads to growth suppression both in vitro and in vivo. We previously determined that changes in MnSOD expression had bidirectional effects on adriamycin (ADR) when combined with nitric oxide (NO). Radiation induces free radicals in a manner similar to ADR, so we speculated that MnSOD combined with NO would also have a bidirectional effect on cellular radiosensitivity. To examine this hypothesis, TE-1 human esophageal squamous carcinoma cells were stably transfected using lipofectamine with a pLenti6-DEST plasmid containing human MnSOD cDNA at moderate to high overexpression levels or with no MnSOD insert. Blastidicin-resistant colonies were isolated, grown, and maintained in culture. We found that moderate overexpression of MnSOD decreased growth rates, plating efficiency, and increased apoptosis. However, high overexpression increased growth rates, plating efficiency, and decreased apoptosis. When combined with NO, moderate overexpression of MnSOD increased the radiosensitivity of esophageal cancer cells, whereas high MnSOD overexpression had the opposite effect. This finding suggests a potential new method to kill certain radioresistant tumors and to provide radioresistance to normal cells.

Inhibitory Effects of Yongbu-tang on Osteoclast Differentiation and Bone Resorption (용부탕의 파골세포 분화 억제와 골 흡수 억제효과)

  • Lee, Jeong Ju;Jo, So Hyun;Park, Min Cheol;Jo, Eun Heui
    • Journal of Acupuncture Research
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    • v.32 no.3
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    • pp.27-40
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    • 2015
  • Objectives : This study was performed to evaluate the effects of water extract of Cervi Parvum Cornu(CPC), Aconiti Lateralis Radix Preparata(ALR), and Yongbu-tang(YBT) on suppression of the receptor activator of nuclear factor kappa-B ligand(RANKL)-induced osteoclast differentiation and bone resorption. Methods : The effects of CPC, ALR, YBT extracts on osteoclast differentiation were determined by culture of bone marrow macrophage(BMM). The mRNA expression levels of the nuclear factor of activated T-cells cytoplasmic 1(NFATc1), c-Fos and tartrate-resistant acid phosphatase(TRAP) in BMMs were analyzed by reverse transcriptase polymerase chain reaction(RT-PCR). Similarly, the protein expression levels of NFATc1, c-Fos, mitogen-activated protein kinase(MAPK)s and ${\beta}$-actin in cell lysates were measured by western blotting. In addition, effects of CPC, ALR and YBT extracts were determined by means of Lipopolysaccharide(LPS)-induced bone-loss with mice. Results : CPC, ALR and YBT extracts showed remarkable inhibition on RANKL-induced osteoclast differentiation without cytotoxicity. CPC and ALR extracts significantly reduced the protein expression level of NFATc1. YBT extract significantly reduced the mRNA expression levels of c-Fos, NFATc1 and the protein expression levels of c-Fos, NFATc1, AKT, p38, c-Jun N-terminal kinase(JNK). Further, YBT extract suppressed degradation of$ I-{\kappa}B$. And ALR extract significantly restored the bone erosion by LPS treatment in mice. Conclusions : YBT extract showed more remarkable inhibition on osteoclast differentiation than CPC and ALR extracts in vitro. ALR extract showed remarkable inhibition on bone resorption in vivo. Thus, YBT extract can be a useful treatment for bone-loss diseases such as osteoporosis.

The Study on Antithrombosis and Inflammation according to The Broth Preparation Method of Gamijoukyungtang (가미조경탕(加味調經湯)의 전탕(煎湯) 방법에 따른 항혈전 및 염증에 관한 연구)

  • Ahn, Kyu-Hwan;Choe, Chang-Min;Kim, Song-Baeg;Cho, Han-Baek
    • The Journal of Korean Obstetrics and Gynecology
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    • v.22 no.1
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    • pp.53-78
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    • 2009
  • Purpose: In this study, we investigated the anti-thrombotic and antiinflammatory efficacy of "Gamijoukyungtang(GJKT)". Methods: We studied inhibitory effects of platelet aggregation, FXa activation, $TXB_2$ and $PGE_2$ biosynthesis and suppressive effects of GPIIb/IIIa activity and oxidative damage, pro-inflammatory cytokine reduction effects of 'GJKT(by press extractor)/GJKT-1(by pressless extractor)' in vitro. Also, we studied suppression of pulmonary embolism, AV shunt model in rats and shortening of Rat tail bleeding time in vivo. Results: GJKT/GJKT-1 extract showed inhibitory effects on GPIIb/IIIaactivities and platelet aggregation induced by ADP, epinephrine, collagen and arachidonic acid. They suppressed biosynthesis of $PGE_2$ but GJKT-1 only supressed biosynthesis of $TXB_2$. In FXa assay, they inhibited activation of FXa. they suppressed pulmonary embolism triggered by collagen and epinephrine. In AV shunt model, they decreased the weights of AV shunt thrombus. they inhibited pro-inflammatory cytokines and decreased oxidative damages caused by DPPH. Conclusion: We confirmed the anti-thrombosis, and ant-inflammatory efficacy of 'GJKT(by press extractor)/GJKT-1(by pressless extractor)'.

Boeravinone B, a natural rotenoid, inhibits osteoclast differentiation through modulating NF-κB, MAPK and PI3K/Akt signaling pathways

  • Xianyu Piao;Jung-Woo Kim;Moonjung Hyun;Zhao Wang;Suk-Gyun Park;In A Cho;Je-Hwang Ryu;Bin-Na Lee;Ju Han Song;Jeong-Tae Koh
    • BMB Reports
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    • v.56 no.10
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    • pp.545-550
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    • 2023
  • Osteoporosis is a major public health concern, which requires novel therapeutic strategies to prevent or mitigate bone loss. Natural compounds have attracted attention as potential therapeutic agents due to their safety and efficacy. In this study, we investigated the regulatory activities of boeravinone B (BOB), a natural rotenoid isolated from the medicinal plant Boerhavia diffusa, on the differentiation of osteoclasts and mesenchymal stem cells (MSCs), the two main cell components responsible for bone remodeling. We found that BOB inhibited osteoclast differentiation and function, as determined by TRAP staining and pit formation assay, with no significant cytotoxicity. Furthermore, our results showing that BOB ameliorates ovariectomy-induced bone loss demonstrated that BOB is also effective in vivo. BOB exerted its inhibitory effects on osteoclastogenesis by downregulating the RANKL/RANK signaling pathways, including NF-κB, MAPK, and PI3K/Akt, resulting in the suppression of osteoclast-specific gene expression. Further experiments revealed that, at least phenomenologically, BOB promotes osteoblast differentiation of bone marrow-derived MSCs but inhibits their differentiation into adipocytes. In conclusion, our study demonstrates that BOB inhibits osteoclastogenesis and promotes osteoblastogenesis in vitro by regulating various signaling pathways. These findings suggest that BOB has potential value as a novel therapeutic agent for the prevention and treatment of osteoporosis.