To investigate the expression of the subunit p65 of NF-kappaB and inhibitor kappa B alpha (IkappaBalpha) in mouse uterus during peri-implantation, thereby investigating whether transient activation of nuclear factor-kappaB (NF-kappaB) takes place during embryo implantation in mice. Immunohistochemical technique was used to examine the expression and localization of p65 in endometrium or deciduas, and Western blot analysis was employed to detect the levels of IkappaBalpha protein in mouse endometrium or deciduas. P65 protein was detected in stromal cells, epithelial cells of endometrium as well as in myometrium. Staining was predominately seen in the cytoplasm of the cells. Staining intensity for p65 was stronger in the epithelial compartment than the stromal compartment and myometrium. Staining intensity increased slightly during pregnancy, and it reached a high level on pregnancy day 5 and day 8. In contrast to p65, the level of IkappaBalpha protein was lowest on pregnancy day 5 in all groups. Our results suggested that NF-kappaB may regulate embryo implantation by its transient activation in mice.
Decidua/metabolism ; Embryo Implantation/*physiology ; Endometrium/metabolism ; I-kappa B Proteins/*biosynthesis ; NF-kappa B/*biosynthesis ; Time Factors ; Uterus/*metabolism ; Uterus/physiology

The inhibitive effects of all-trans retinoic acid (ARTA) on airway inflammation in asthmatic rats and its mechanism on the basis of the regulation of nuclear factor kappaB (NF-kappaB) were explored. Thirty-two SD rats were randomly divided into 4 groups: control group, asthma group, dexamethasone treatment group and retinotic acid treatment group. The total and differential cell counts in the collected bronchoalveolar lavage fluid (BALF) were measured. The pathological changes in lung tissues were estimated by scoring. The expression of NF-kappaB inhibitor (IkappaBa), NF-kappaB, intercellular adhering molecule-1 (ICAM-1) in lung tissue was detected by immunohistochemical method. The results showed that in the two treatment groups, the total cell counts and proportion of inflammatory cells in BALF were significantly reduced, but there was no significant difference in differential cell counts in BALF between them. The pathological changes in lung tissues in the treatment groups were significantly attenuated as compared with asthma group. Except the epithelial injury in retinotic acid treatment group was milder than in dexamethasone treatment group, the remaining lesions showed no significant difference between them. In the two treatment groups, the expression of IkappaBa was increased, while the expression of NF-kappaB and ICAM-1 decreased with the difference between the two groups being not significant. It was concluded that the similar anti-inflammatory effects and mechanism of ATRA on airway in asthmatic rats to those of dexamethasone were contributed to the increase of cytoplasmic IkappaBa content and suppression of NF-kappaB activation and expression.
Animals ; Asthma ; metabolism ; pathology ; Bronchoalveolar Lavage Fluid ; cytology ; Dexamethasone ; pharmacology ; I-kappa B Proteins ; biosynthesis ; genetics ; Inflammation ; Intercellular Adhesion Molecule-1 ; biosynthesis ; genetics ; Male ; NF-kappa B ; biosynthesis ; genetics ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Respiratory System ; Tretinoin ; pharmacology

To investigate the expression of the subunit p65 of NF-kappaB and inhibitor kappa B alpha (IkappaBalpha) in mouse uterus during peri-implantation, thereby investigating whether transient activation of nuclear factor-kappaB (NF-kappaB) takes place during embryo implantation in mice. Immunohistochemical technique was used to examine the expression and localization of p65 in endometrium or deciduas, and Western blot analysis was employed to detect the levels of IkappaBalpha protein in mouse endometrium or deciduas. P65 protein was detected in stromal cells, epithelial cells of endometrium as well as in myometrium. Staining was predominately seen in the cytoplasm of the cells. Staining intensity for p65 was stronger in the epithelial compartment than the stromal compartment and myometrium. Staining intensity increased slightly during pregnancy, and it reached a high level on pregnancy day 5 and day 8. In contrast to p65, the level of IkappaBalpha protein was lowest on pregnancy day 5 in all groups. Our results suggested that NF-kappaB may regulate embryo implantation by its transient activation in mice.
Animals ; Decidua ; metabolism ; Embryo Implantation ; physiology ; Endometrium ; metabolism ; Female ; I-kappa B Proteins ; biosynthesis ; Mice ; NF-KappaB Inhibitor alpha ; NF-kappa B ; biosynthesis ; Pregnancy ; Time Factors ; Uterus ; metabolism ; physiology

<b>OBJECTIVEb>To evaluate the role of nuclear factor-kappa B (NF-kappaB) and inhibitory kappaB alpha (IkappaBalpha) in hepatocellular cacinoma (HCC) SMMC7721 cells, the consequence of NF-kappaB inhibition in SMMC7721 cells transfected with mutated IkappaBalpha (mIkappaBalpha) plasmid and the effect of stable inhibition of NF-kappaB activity in combination with Doxorubicin.

<b>METHODSb>Western blot was used to determine the expression of NF-kappaB and IkappaBalpha in SMMC7721 cells and normal liver cells. Nuclear protein was used to evaluate the binding of the (32)P-labeled tandem kappaB sequence using electrophoretic mobility shift assay and the expression of NF-kappaB using Western blot between SMMC7721 cells transfected with mIkappaBalpha plasmid (SMMC7721-MT) and control cells. Furthermore, cell viability was plotted between SMMC7721-MT and control cells. The binding of kappaB sequence and cell viability between SMMC7721-MT and control cells at different concentrations of Doxorubicin were also investigated.

<b>RESULTSb>Western blot analysis for nuclear extract showed more P50 (NF-kappaB1) and P65 (RelA) expression in SMMC7721 cells compared with normal liver cells. The expression of cytosolic IkappaBalpha protein in SMMC7721 cells was less than that in normal cells. SMMC7721-MT cells inhibited NF-kappaB nuclear translocation at 0, 24, 48 and 96 hours. Furthermore, NF-kappaB cannot be detected in the nuclear protein of SMMC7721-MT cells by Western blot. By calculating cell viability, the proliferation of SMMC7721-MT cells was shown to be suppressed more significantly than that of control cells. NF-kappaB in untransfected cells was activated by Doxorubicin in a dose-dependent manner, but that in SMMC7721-MT cells was not induced at low concentrations of Doxorubicin. Compared with untransfected cells, the viability of SMMC7721-MT cells was significantly suppressed at the same concentration of Doxorubicin (P < 0.01).

<b>CONCLUSIONSb>The present study demonstrates that upregulation of NF-kappaB and downregulation of inhibitory kappa B (IkappaBalpha) in SMMC7721 cells are related with the growth of hepatocellular cacinoma cells. Stable expression of mIkappaBalpha in SMMC7721-MT cells can inhibit NF-kappaB nuclear translocation and suppress cell growth. Furthermore, stable inhibition of NF-kappaB activity in combination with Doxorubicin can significantly inhibit cell proliferation in SMMC7721-MT cells. Thus, modulation of NF-kappaB may represent an improvement in the efficacy of HCC therapies and be worthy of further research and investigation.

Antineoplastic Agents ; pharmacology ; Blotting, Western ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Division ; Doxorubicin ; pharmacology ; Electrophoretic Mobility Shift Assay ; Humans ; I-kappa B Proteins ; biosynthesis ; Liver Neoplasms ; metabolism ; pathology ; NF-kappa B ; biosynthesis ; physiology ; Tumor Cells, Cultured

<b>OBJECTIVEb>To investigate the effects of oxymatrine on the expression of nuclear factor kappa B (NF-kappaB) in the kidneys of rats with adriamycin-induced chronic renal fibrosis.

<b>METHODSb>Totally 120 Wistar rats were randomly assigned to normal control group, renal fibrosis model group, benazepril treatment group and oxymatrine treatment group (n=30). The rats in the normal control were injected with normal saline via the tail vein, and those in the other 3 groups with adriamycin (2 mg/kg) on the 7th day and 21st day of the experiment, respectively. Oxymatrine (100 mg/kg) or benazepril (6 mg/kg) was given by gastric perfusion after the second injection. Every 4 weeks after the second injection, 5 rats in each group were killed to evaluate the pathological changes and functional impairment of the kidney. Immunohistochemistry was used to detect the expression of NF-kappaB and inhibitory kappa B (IkappaB) in the kidney. The association of NF-kappaB expression with IkappaB expression, renal pathological changes and functional impairment were studied.

<b>RESULTSb>Oxymatrine and benazepril ameliorated renal fibrosis and functional impairment. Immunohistochemical staining revealed increased NF-kappaB expression and decreased IkappaB expression in the model group in comparison with oxymatrine and benazepril treatment groups 8 weeks after the second injection, but no significant difference was noted between the latter two groups. NF-kappaB expression in the kidneys of rats with adriamycin-induced chronic renal fibrosis showed an inverse correlation with IkappaB expression and positive correlation with pathological changes and functional impairment.

<b>CONCLUSIONb>Oxymatrine may inhibit renal fibrosis by down-regulating NF-kappaB expression, which may play a key role in protection against renal fibrosis.

Alkaloids ; pharmacology ; Animals ; Chronic Disease ; Doxorubicin ; Fibrosis ; chemically induced ; I-kappa B Proteins ; biosynthesis ; Immunohistochemistry ; Kidney ; drug effects ; metabolism ; pathology ; Male ; NF-kappa B ; biosynthesis ; Quinolizines ; pharmacology ; Random Allocation ; Rats ; Rats, Wistar

The involvement of NF-kappaB binding activity is known to be important in the mechanism of acute liver injury and in the induction of cyclooxygenase (COX-2). This study was performed to evaluate NF-kappaB binding activity and the expression of COX-2 in chronic liver injury induced by carbon tetrachloride (betaCCI(4)). Liver tissues from Sprague - Dawley rats were collected at 1, 3, 5, and 7th week after intraperitoneal injection of 0.1 mL of betaCCI(4)/100 g body weight twice a week. Reactive oxy-gen species (ROS) were measured in the postmitochondrial fraction by dichlorofluorescein formation with a fluorescent probe. An electrophoretic mobility shift assay was performed for NF-kappaB binding activity. Western blot was performed to measure the level of COX-1, COX-2, p65, p50, and I B proteins. ROS and NF-kappaB activity increased during the CCl4-induced chronic liver injury. The expression of nuclear p65 protein and p50 protein increased compared with that of the control, while the cytoplasmic I B protein decreased as the inflammation persisted. The expression of COX-2 in betaCCI(4)-treated rat liver increased compared with that of the control. It could be suggested that ROS produced by betaCCI(4) treatment increased NF-kappaB binding activity and thereby COX-2 expression, and these might be implicated in the progress of chronic liver damage.
Animals ; Biological Transport ; Carbon Tetrachloride/administration & dosage/*adverse effects ; Carbon Tetrachloride Poisoning/*metabolism/pathology ; Cell Nucleus/metabolism ; Cyclooxygenase 1 ; Cyclooxygenase 2 ; Cytoplasm/metabolism ; I-kappa B Proteins/biosynthesis ; Isoenzymes/*biosynthesis ; Liver/drug effects/*injuries/pathology ; Membrane Proteins ; NF-kappa B/antagonists & inhibitors/*metabolism ; NF-kappa B p50 Subunit ; Prostaglandin-Endoperoxide Synthases/*biosynthesis ; Protein Binding ; Rats ; Rats, Sprague-Dawley ; Reactive Oxygen Species ; Transcription Factor RelA

<b>OBJECTIVEb>To investigate the inhibition consequence of NF-kappaB activity and cell viability by transfecting mutated inhibitor kappa B alpha (mI(kappa)B(alpha)) into liver cancer cell line of SMMC-7721 cells.

<b>METHODSb>The nucleic proteins of SMMC-7721 cells transfected with mI(kappa)B(alpha) plasmid and cells with empty pcDNA3 vector were used to determine not only the binding of the 32P-labelled kappaB probes by EMSA, but also the expression of NF-kappaB by western blot. Cell viability was also analyzed.

<b>RESULTSb>NF-kappaB nuclear translocation was inhibited remarkably in SMMC-7721 cells transfected with mI(kappa)B(alpha) at 0, 24, 48 and 96 hours. Furthermore, NF-kappaB was not detected in the nucleic protein of mI(kappa)B(alpha) -transfected cells at the same intended time by western blot. Compared with that of control cells, the growth of SMMC-7721 cells transfected with mI(kappa)B(alpha) was suppressed evidently, especially on the second day, the cpm values of mI(kappa)B(alpha) -transfected cells, pcDNA3-transfected cells, and control cells were 5,092.63+/-541.41, 7,851.87+/-72.76, and 8,240.8+/-603.26 respectively (t = 14.29, P<0.01; t = 10.99, P<0.01).

<b>CONCLUSIONb>Stable expression of mI(kappa)B(alpha) in SMMC-7721 cells transfected with mI(kappa)B(alpha) plasmid inhibits NF-kappaB nuclear translocation, then suppresses the cell growth.

Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Division ; Cell Line, Tumor ; Humans ; I-kappa B Proteins ; biosynthesis ; genetics ; physiology ; Liver Neoplasms ; metabolism ; pathology ; Mutation ; NF-KappaB Inhibitor alpha ; NF-kappa B ; antagonists & inhibitors ; physiology ; Transfection ; Translocation, Genetic

<b>OBJECTIVEb>To investigate the effect of homocysteine (HCY) on activation of nuclear factor (NF-kappaB) and inhibitory factor IkappaB-alpha in human monocyte cell line THP-1, as well as its association with macrophage inflammatory protein (MIP-1alpha) upregulation.

<b>METHODSb>THP-1 monocytes were incubated with HCY, with and without NF-kappaB inhibitor pyrolidine dithiocarbamate (PDTC) pretreatment. Northern blot analysis and flow cytometry were used to detect MIP-1alpha mRNA and protein respectively. The nuclear protein NF-kappaB P65 subunit and the inhibitory protein IkappaB-alpha were analyzed by Western blotting.

<b>RESULTSb>Compared with controls, HCY, at a concentration of 0.1 mmol/L, was able to enhance the expression of MIP-1alpha mRNA (up to 3.69-fold) and protein (1.16-fold) in THP-1 monocytes, as well as enhance NF-kappaB P65 transcription to nuclear proteins. These actions were significantly suppressed after pretreatment with 100 micromol/L PDTC for 30 minutes before HCY incubation; whereas incubation of THP-1 monocytes with PDTC only had no effect on both the expression of MIP-1alpha and nuclear transcription of NF-kappaB P65. Moreover, the level of IkappaB-alpha protein in THP-1 monocytes decreased after a 30-minute incubation with HCY, which gradually increased after 120 minutes.

<b>CONCLUSIONSb>Homocysteine at a pathologic concentration stimulates MIP-1alpha expression in THP-1 monocytes, probably via NF-kappaB activation. Such activation may be caused by enhanced phosphorylation and degradation of the inhibitor protein IkappaB-alpha.

Cell Line, Tumor ; Chemokine CCL3 ; Chemokine CCL4 ; Homocysteine ; pharmacology ; Humans ; I-kappa B Proteins ; metabolism ; Leukemia, Monocytic, Acute ; metabolism ; pathology ; Macrophage Inflammatory Proteins ; biosynthesis ; genetics ; Monocytes ; metabolism ; NF-KappaB Inhibitor alpha ; NF-kappa B ; antagonists & inhibitors ; Phosphorylation ; Proline ; analogs & derivatives ; pharmacology ; RNA, Messenger ; biosynthesis ; genetics ; Thiocarbamates ; pharmacology ; Transcription Factor RelA ; biosynthesis ; genetics ; Transcription, Genetic

In order to investigate the effect of nuclear factor-kappa B (NF-kappaB) on vascular endothelial growth factor (VEGF) mRNA expression of human pulmonary artery smooth muscle cells (HPASMCs) in hypoxia, the cultured HPASMCs in vitro were stimulated with pyrrolidine dithiocarbamate (PDTC), an inhibitor of NF-kappaB. The NF-kappaB p65 nuclei positive expression was detected by immunocytochemical technique. The IkappaBalpha protein expression was measured by Western blot. RT-PCR was used to detect the VEGF mRNA expression of HPASMCs. The results showed that no significant change was observed in the NF-kappaB p65 nuclei positive expression of cultured HPASMCs during 6 h-24 h in normoxia, but the levels of NF-kappaB p65 nuclei positive expression of cultured HPASMCs were significantly increased in hypoxia groups as compared with those in all normoxia groups (P<0.05). The IkappaBalpha protein expression of cultured HPASMCs showed no significant change during 6 h-24 h in normoxia, but significantly decreased in hypoxia as comapred with that in normoxia groups (P<0.05). PDTC (1 to 100 micromol/L) could inhibit the VEGF mRNA expression of HPASMCs in a concentration-dependent manner in hypoxia. In conclusion, NF-kappaB can be partly translocation activated from cytoplasm into nuclei in the cultured HPASMCs under hypoxia. The inhibition of NF-kappaB activation can decrease the VEGF mRNA expression. It is suggested that the activation of NF-kappaB is involved in the VEGF mRNA expression of HPASMCs under hypoxia.
Cell Hypoxia ; Cells, Cultured ; Humans ; I-kappa B Proteins ; metabolism ; Muscle, Smooth, Vascular ; cytology ; metabolism ; Myocytes, Smooth Muscle ; cytology ; metabolism ; NF-kappa B ; metabolism ; Pulmonary Artery ; cytology ; Pyrrolidines ; pharmacology ; RNA, Messenger ; biosynthesis ; genetics ; Thiocarbamates ; pharmacology ; Transcription Factor RelA ; Vascular Endothelial Growth Factor A ; biosynthesis ; genetics

<b>OBJECTIVEb>To explore the apoptosis induction by curcumin in androgen-dependent prostate cancer cell line LNCaP).

<b>METHODSb>After LNCaP cells were induced by 10, 25, 50, 75, 100 micromol/L curcumin respectively, the cell activity was assayed by MTT at 5, 12 and 24 hours. Flow cytometry and electronic microscopy were adopted to observe cell cycle and morphological changes of LNCaP cells at 24 hours. After 5 hours, the expression of IkappaBalpha in LNCaP cells was detected by Western blotting.

<b>RESULTSb>The growth of LNCaP cells was suppressed obviously by curcumin in dose-dependent and time-dependent manners in vitro. There were significant differences in inhibition rate among different concentrations and time groups (P < 0.05). Furthermore, curcumin could arrest the cell cycle of LNCaP cells at G2/M phase in a dose-dependent manner (P <0.01). The ratios of apoptosis were significantly higher than those of controls (P < 0. 5). Curcumin could lead to characteristic morphological changes of apoptosis in LNCaP cells after 24 hours. The expression of IkappaBalpha in LNCaP cell did not show marked changes after the exposure to different concentrations of curcumin within 5 hours.

<b>CONCLUSIONb>Curcumin can suppress the growth of LNCaP, and promotes their apoptosis.

Apoptosis ; drug effects ; Cell Cycle ; Cell Line, Tumor ; Cell Proliferation ; Curcumin ; pharmacology ; Dose-Response Relationship, Drug ; Humans ; I-kappa B Proteins ; biosynthesis ; Male ; NF-KappaB Inhibitor alpha ; Neoplasms, Hormone-Dependent ; Prostatic Neoplasms ; metabolism ; pathology