1.Expression of gene associated with retinoid-interferon-induced mortality-19 in preimplantation embryo of mice.
Wen-juan CUI ; Lan CHAO ; Xiao-hui DENG ; Yan-jun SHEN ; Fang YANG ; Wen-juan FENG ; Jing XU ; Hong-lei CHEN
Acta Academiae Medicinae Sinicae 2012;34(3):212-215
OBJECTIVETo study the expression of gene associated with retinoid-interferon-induced mortality-19(GRIM-19) in preimplantation embryo of mice and explore its role in embryonic development.
METHODSThe protein and mRNA expressions of GRIM-19 in 2-cell, 4-cell, 8-cell, morula, and blastocyst phases of mice preimplantation embryo were detected by Western blot analysis and Real-time polymerase chain reaction (PCR).
RESULTSGRIM-19 was continuously expressed in every stage of preimplantation embryo of mice. Western blot analysis and Real-time PCR demonstrated a gradual increase of GRIM-19 expression from 2-cell, which reached a peak in 8-cell phase and then decreased progressively.
CONCLUSIONSThe expression of GRIM-19 in mouse preimplantation embryos changes as at different developmental phases. GRIM-19 may play an important role during embryonic development.
Animals ; Blastocyst ; drug effects ; metabolism ; Female ; Interferons ; pharmacology ; Mice ; NADH, NADPH Oxidoreductases ; genetics ; metabolism ; Pregnancy ; RNA, Messenger ; genetics ; Tretinoin ; pharmacology
2.Effect of water deficit on gene expression of enzymes related with hydrogen peroxide detoxification system in Scutellaria baicalensis.
Chong WU ; Shuangshuang QIN ; Yuan YUAN ; Ping CHEN ; Shuifang LIN
China Journal of Chinese Materia Medica 2012;37(2):186-188
OBJECTIVETo analysis the effects of water deficit on the transcript level of SOD, APX, DHAR and MDHAR genes in Scutellaria baicalensis.
METHODThree-month-old S. baicalensis was in glasshouse under water deficit stress, and the transcript level of SOD, APX, DHAR and MDHAR genes were analysis utilized semi-quantitative RT-PCR.
RESULTCompared with the control group, a significant decline of the transcriptional level of APX gene was observed at 70 days after water deficit. The transcript level of DHAR gene was reduced at 30 and 50 days after water deficit. And MDHARI gene was significant declined at 50 days.
CONCLUSIONAsA which is an important antioxidant plays a major role in hydrogen peroxide clear system under water deficit, and maybe have an antagonistic effect to the accumulation of baicalein.
Ascorbate Peroxidases ; genetics ; metabolism ; Ascorbic Acid ; metabolism ; Gene Expression Regulation, Enzymologic ; drug effects ; Gene Expression Regulation, Plant ; drug effects ; Hydrogen Peroxide ; metabolism ; NADH, NADPH Oxidoreductases ; genetics ; metabolism ; Oxidoreductases ; genetics ; metabolism ; Plant Proteins ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Scutellaria baicalensis ; enzymology ; genetics ; metabolism ; Superoxide Dismutase ; genetics ; metabolism ; Time Factors ; Water ; metabolism ; pharmacology
3.Inhibition of rac1 Reduces PDGF-induced Reactive Oxygen Species and Proliferation in Vascular Smooth Muscle Cells.
Gu KONG ; Sahng LEE ; Kyung Soo KIM
Journal of Korean Medical Science 2001;16(6):712-718
In vascular smooth muscle cells, reactive oxygen species (ROS) were known to mediate platelet-derived growth factor (PDGF)-induced cell proliferation and NADH/NADPH oxidase is the major source of ROS. NADH/NADPH oxidase is controlled by rac1 in non-phagocytic cells. In this study, we examined whether the inhibition of rac1 by adenoviral-mediated gene transfer of a dominant negative rac1 gene product (Ad.N17rac1) could reduce the proliferation of rat aortic vascular smooth muscle cells (RASMC) stimulated by PDGF via decreasing intracellular ROS. RASMC were stimulated by PDGF (80 ng/mL) with or without N-acetylcysteine 1 mM or infected with 100 mutiplicity of infection of Ad.N17rac1. Intracellular ROS levels were measured at 12 hr using carboxyl-2', 7'-dichlorodi-hydrofluorescein diacetate confocal microscopy. At 72 hr, cellular proliferation was evaluated by cell number counting and XTT assay. Compared with control, ROS levels were increased by 2-folds by PDGF. NAC and Ad.N17rac1 inhibited PDGF-induced increase of ROS by 77% and 65%, respectively. Cell number was increased by PDGF by 1.6-folds compared with control. NAC and Ad.N17rac1 inhibited PDGF-induced cellular growth by 45% and 87%, respectively. XTT assay also showed similar results. We concluded that inhibition of rac1 in RASMCs could reduce intracellular ROS levels and cellular proliferation induced by PDGF.
Adenoviridae/genetics
;
Animal
;
Aorta, Thoracic/cytology
;
Cell Division/drug effects/physiology
;
Cells, Cultured
;
Gene Expression/physiology
;
Gene Transfer Techniques
;
Multienzyme Complexes/antagonists & inhibitors
;
Muscle, Smooth, Vascular/*cytology/*metabolism
;
NADH, NADPH Oxidoreductases/antagonists & inhibitors
;
NADPH Oxidase/antagonists & inhibitors
;
Platelet-Derived Growth Factor/*pharmacology
;
Rats
;
Rats, Sprague-Dawley
;
Reactive Oxygen Species/*metabolism
;
rac1 GTP-Binding Protein/*genetics/metabolism
4.Resveratrol inhibits foam cell formation via NADPH oxidase 1-mediated reactive oxygen species and monocyte chemotactic protein-1.
Dae Weon PARK ; Kheewoong BAEK ; Jae Ryong KIM ; Jae Jin LEE ; Sang Ho RYU ; Byung Rho CHIN ; Suk Hwan BAEK
Experimental & Molecular Medicine 2009;41(3):171-179
Resveratrol is a polyphenolic compound in red wine that has anti-oxidant and cardioprotective effects in animal models. Reactive oxygen species (ROS) and monocyte chemotactic protein-1 (MCP-1) play key roles in foam cell formation and atherosclerosis. We studied LPS-mediated foam cell formation and the effect of resveratrol. Resveratrol pretreatment strongly suppressed LPS-induced foam cell formation. To determine if resveratrol affected the expression of genes that control ROS generation in macrophages, NADPH oxidase 1 (Nox1) was measured. Resveratrol treatment of macrophages inhibited LPS-induced Nox1 expression as well as ROS generation, and also suppressed LPS-induced MCP-1 mRNA and protein expression. We investigated the upstream targets of Nox1 and MCP-1 expression and found that Akt-forkhead transcription factors of the O class (FoxO3a) is an important signaling pathway that regulates both genes. These inhibitory effects of resveratrol on Nox1 expression and MCP-1 production may target to the Akt and FoxO3a signaling pathways.
Antioxidants/*pharmacology
;
Cells, Cultured
;
Chemokine CCL2/genetics/*metabolism
;
Enzyme Activation/drug effects
;
Foam Cells/*drug effects/physiology
;
Forkhead Transcription Factors/metabolism
;
Humans
;
Lipopolysaccharides/pharmacology
;
NADH, NADPH Oxidoreductases/genetics/*metabolism
;
Proto-Oncogene Proteins c-akt/metabolism
;
RNA, Messenger/metabolism
;
Reactive Oxygen Species/*metabolism
;
Signal Transduction
;
Stilbenes/*pharmacology