OBJECTIVE: To study the expression and activation of p50 subunit of nuclear factor-kappa B(NF-kappaB) in mucosa of seasonal allergic rhinitis. METHOD: The expression of p50 subunit of NF-kappaB in the mucosa from 16 patients (6 patients with symptom, 10 patients without symptom)and 10 normal subjects were detected by immunohistochemistry. The activation of DNA-binding proteins which was labeled with 32P-radiolabeled oligonucleotide probe for NF-kappaB was detected with electrophoretic mobility shift assays(EMSA) in mucosa. RESULT: The expression of p50 subunit of NF-kappaB was observed in the nasal mucosa of SAR and normal samples. The expression of p50 subunit of NF-kappaB was positive in the cytoplasm and some nuclei of the mucosal epithelia, inflammatory cells, glandular epithelia, and vascular endothelia in nasal mucosa. The Rate of nucleus positive staining of p50 was (41.83 +/- 4.43)% and(37.19 +/- 3.93)% in SAR with symptom and SAR without symptom patients,respectively. There was no significant difference (P > 0.05). The Rate of nucleus positive staining of p50 in nasal mucosa of normal samples was (8.89 +/- 1.32)%. The difference of p50 subunit of NF-kappaB expression between SAR group and normal group was statistically significant (P < 0.01). The DNA-binding proteins activity of samples from patients with seasonal allergic rhinitis was stronger than that in normal subjects (P < 0.05). CONCLUSION p50 subunit of NF-kappaB was activated in healthy nasal mucosa to some extent. The expression and DNA-binding proteins activity of p50 subunit of NF-kappaB was enhanced in seasonal allergic rhinitis. It indicated that p50 subunit of NF-kappaB may be involved in nasal mucosa physiological function and may have an important role in maintaining the chronic inflammation in seasonal allergic rhinitis.
Case-Control Studies ; Humans ; NF-kappa B p50 Subunit ; genetics ; metabolism ; Nasal Mucosa ; metabolism ; Rhinitis, Allergic, Seasonal ; metabolism

Objective: To investigate the regulatory effect of the transcription factor NF-kB1 on the expression of miR-195 in prostate cancer (PCa). METHODS: We analyzed the possibility of NF-kB1 binding to the miR-195 promoter and the expression of NF-kB1 in PCa using the JASPAR and Oncomine databases, respectively, and determined the expressions of NF-kB1 and miR-195 in PCa cells by real-time quantitative PCR after inhibiting the former by interfering RNA targeting NF-kB1. We detected the activity of the luciferase reporter gene after constructing its gene plasmid in the miR-195 promoter region and having it co-transfected with the NF-kB1 plasmid. Then we analyzed the correlation between the expressions of miR-195 and NF-kB1 in the prostate tissue. RESULTS: NF-kB1 was overexpressed in PCa. After inhibition of the expression of NF-kB1, that of miR-195 was increased in PC-3 and DU-145 cell lines, with a negative correlation between the NF-kB1 and miR-195 expressions in the PCa tissue. The results of luciferase reporter gene assay showed direct binding of NF-kB1 to the miR-195 promoter zone. CONCLUSIONS NF-kB1 regulates the expression of miR-195 in prostate cancer.
Cell Line, Tumor ; Gene Expression Regulation, Neoplastic ; Humans ; Male ; MicroRNAs/genetics* ; NF-kappa B p50 Subunit/metabolism* ; Promoter Regions, Genetic ; Prostatic Neoplasms/genetics* ; Transcription Factors/metabolism*

<b>OBJECTIVEb>To explore how NF-&kappa;B family members regulate maspin expression in prostate cancer cells.

<b>METHODSb>The expression of NF-&kappa;B subunits and maspin was detected by Western blot analysis in prostate cancer DU145, PC-3, and LNCaP cell lines. RNA interference was performed to analyze whether RelB- or RelA-deletion affectes cell death as well as the expression of NF-&kappa;B subunits and maspin. The impact of RelB-silencing in DU145 cells was investigated by flow cytometry. The regulation of RelB on maspin expression in the prostate cancer PC-3 cells was also examined via stable transfection of RelB expression plasmid.

<b>RESULTSb>RelA, p50, RelB, and p52 were constitutively expressed in androgen-independent prostate cancer DU145 and PC-3 cells, while RelB had the highest expression in DU145 cells. Low expression of maspin was detected in LNCaP and DU145 cells, but elevated expression in PC-3 cells. RelB-silencing in DU145 cells by siRNA interference upregulated the endogenous expression of maspin and induced cell apoptosis (13.3±4.2)%. Overexpression of RelB in PC-3 cells inhibited the endogenous expression of maspin. RelA-silecing had no significant influence on the endogenous expression of maspin.

<b>CONCLUSIONSb>The classical and alternative NF-&kappa;B activitions are sustained in androgen-independent prostate cancer cell lines. The expressions of RelB and maspin are inversely correlated in these cancer cells. The expression of RelB negatively regulates the endogenous expression of maspin, then interferes the cell survival. RelA is not involved in the regulation of maspin expression.

Apoptosis ; Cell Line, Tumor ; Gene Silencing ; Humans ; Male ; NF-kappa B ; genetics ; metabolism ; NF-kappa B p50 Subunit ; genetics ; metabolism ; NF-kappa B p52 Subunit ; genetics ; metabolism ; Prostatic Neoplasms ; genetics ; metabolism ; pathology ; RNA, Small Interfering ; genetics ; Serpins ; metabolism ; Transcription Factor RelA ; genetics ; metabolism ; Transcription Factor RelB ; genetics ; metabolism ; Transfection

<b>OBJECTIVEb>To investigate the correlation between nuclear factor-kappa B (NF-kappaB) activity and cytokine expression in nasal mucosa of chronic sinusitis.

<b>METHODSb>IL-5, IL-6 and IL-8 levels in nasal mucosa were assayed by the method of ELISA in 52 cases of chronic sinusitis [concomitant with allergic rhinitis (AR group), without allergic rhinitis (NAR group)] and 12 normal subjects. Semi-quantitative RT-PCR and immunohistochemical staining were used to examine P50 and P65 subunits of NF-KB expressions and activation in nasal mucosa. The correlation between activities of NF-KB P50 and P65 subunits and cytokine expression was evaluated.

<b>RESULTSb>IL-5, IL-6 and IL-8 levels in both AR and NAR groups were significantly increased (all P < 0.01 for AR group; P < 0.05, 0.05, 0.01, respectively, for NAR group, as compared with normal group), and the levels were much higher in AR group than that in NAR group (P < 0.01, 0.05, 0.01, respectively). The levels of P50 and P65 mRNA in both AR and NAR groups were enhanced (all P < 0.01 for AR group; all P < 0.01 for NAR group, as compared with normal group), and AR group had markedly greater P50 and P65 mRNA levels in comparison with NAR group (both P < 0.05). Immunohistochemical study revealed that nucleus-present rates of P50 and P65 in both AR and NAR groups were significantly higher than those of control group (all P < 0.01), and they were much greater in AR group as compared with NAR group (all P < 0.01). Pearson correlation analysis demonstrated that P50 and P65 nucleus-present rates were closely correlated with IL-6 and IL-8 levels, but not IL-5. The correlation coefficient was 0. 49 for P50 and IL-6, 0. 54 for P50 and IL-8, 0. 61 for P65 and IL-6, and 0.66 for P65 and IL-8 (all P < 0.01).

<b>CONCLUSIONSb>Activation of P50 and P65 subunits of NF-kappaB might be one of the mechanisms for induction of IL-6 and IL-8 expression in chronic sinusitis. Concomitance of allergic rhinitis with chronic sinusitis further increased activities of NF-kappaB subunits, and further elevated IL-6 and IL-8 expression. IL-5 expression was independent of NF-kappaB pathway in chronic sinusitis.

Adult ; Chronic Disease ; Female ; Humans ; Interleukin-5 ; metabolism ; Interleukin-6 ; metabolism ; Interleukin-8 ; metabolism ; Male ; Middle Aged ; NF-kappa B p50 Subunit ; metabolism ; Nasal Mucosa ; metabolism ; RNA, Messenger ; genetics ; Rhinitis ; metabolism ; Sinusitis ; metabolism ; Transcription Factor RelA ; metabolism

<b>OBJECTIVEb>The development of bronchopulmonary dysplasia (BPD) is attributed to intrauterine inflammatory and postnatal mechanical ventilation and hyperoxia. The present study was aimed to investigate the effects of lipopolysaccharide (LPS) and hyperoxia exposure on the nuclear factor-kappa B (NF-kappaB) expression in human embryo lung fibroblasts (HELFs) in vitro.

<b>METHODSb>Either LPS (100 ng/mL) or hyperoxia (60%), or a combination of both was employed to stimulate confluent HELFs. After 0.5, 1, 2 and 4 hrs of stimulation, the nuclear translocation of two subunits p50 and p65 in HELFs was detected with immunocytochemistry. Reverse transcription quantitative polymerase chain reaction (RT-PCR) was used to measure mRNA expression of NF-kappaB p50 and p65.

<b>RESULTSb>LPS or hyperoxia stimulation induced the nuclear translocation of p50 and p65 at 30 minutes of exposure. mRNA expression of NF-kappaB p50 and p65 peaked at 1 hr and then gradually decreased. A stimulation of LPS combined with hyperoxia induced the nuclear translocation of p50 and p65. NF-kappaB p50 and p65 mRNA expression peaked at 2 hrs of stimulation and then decreased slowly, but was significantly higher than that in the LPS or hyperoxia stimulation alone group 4 hrs after stimulation.

<b>CONCLUSIONSb>Both LPS and hyperoxia exposure induced NF-kappaB activation in the HELFs in vitro. Hyperoxia combined with LPS induced a more prolonged duration of NF-kappaB activation. This suggests that the individuals who were subjected to intrauterine inflammation and postnatal hyperoxia exposure are more vulnerable to lung injury.

Bronchopulmonary Dysplasia ; etiology ; Fibroblasts ; metabolism ; Humans ; Hyperoxia ; metabolism ; Immunohistochemistry ; Infant, Newborn ; Lipopolysaccharides ; toxicity ; Lung ; cytology ; embryology ; NF-kappa B p50 Subunit ; analysis ; genetics ; metabolism ; Transcription Factor RelA ; analysis ; genetics ; metabolism

<b>OBJECTIVEb>To investigate the expression and significance of Toll like receptor (TLR)4 mRNA and nuclear factor (NF)-kappaB p50 mRNA in human normal nasal mucosal cell before and after stimulation by LPS.

<b>METHODSb>The tissue was obtained from 15 normal middle turbinates (without rhinosinusitis). Every tissue was cultured in vitro, divided into 2 specimens. LPS was added into 15 specimens as LPS group and not added into other 15 specimens as control group. The pathomorphological characters of nasal mucosal cells were observed under optical microscope after stimulation by LPS. The expression of TLR4 mRNA and NF-kappaB p50 mRNA in normal human nasal mucosal cells were evaluated by in situ hybridization.

<b>RESULTSb>Normal mucociliary agglutinated and mucosal cells were enlarged after stimulation by LPS; The expression of TLR4 mRNA in LPS group was higher than control group obviously. Their average density of light was 1.283 +/- 0.027 in LPS group while 0.538 +/- 0.038 in control group, and there was statistical significance between the two groups (t = 1.761, P < 0.05). The expression of NF-kappaB p50 mRNA was higher than control group obviously, and expressed in cellular nucleus predominantly. Their average density of light was 1. 668 +/- 0.037 in LPS group while 0. 372 +/- 0.052 in control group, and there was statistical significance between the two groups (t = 2. 624, P < 0. 01).

<b>CONCLUSIONSb>LPS can activate the NF-kappaB p50 of human nasal mucosal cells through TLR4, and it may play some roles in stimulating and damage effect induced by LPS in nasal mucosal cells.

Adult ; Epithelial Cells ; metabolism ; Female ; Humans ; In Vitro Techniques ; Lipopolysaccharides ; Male ; Middle Aged ; NF-kappa B p50 Subunit ; metabolism ; Nasal Mucosa ; cytology ; metabolism ; RNA, Messenger ; genetics ; Toll-Like Receptor 4 ; genetics ; metabolism ; Young Adult

OBJECTIVE: To clarify the pathogenesis of allergic rhinitis (AR) by detecting the activation of NF-kappaB and the expression of ICAM-1 mRNA in nasal mucosa of the patients with AR. METHOD: AR patients were collected as experimental group and deflection of nasal septum patients as control group. Immunohistochemistry methods were used to examine the expression of NF-kappaB p50 and p65 subunits in the middle turbinate mucous tunic. ICAM-1 mRNA were detected by means of RT-PCR. RESULT: NF-kappaB was strongly expressed in AR group. NF-kappaB p50 existed in cytoplasm and nucleus. NF-kappaB p65 was mainly expressed in cytoplasm and less in nucleus. While NF-kappaB was expressed little in control group. The difference of NF-kappaB p50 and NF-kappaB p65 expression between two groups were significantly (P < 0.01). ICAM-1 mRNA was more strongly expressed in AR group (P < 0.01) while feebly expressed in deflection of nasal septum group. The expression of ICAM-1 mRNA were significantly correlated with that of NF-kappaB p50 and NF-kappaB p65 (r = 0.899 5, P < 0.01; r = 0.7601, P < 0.01). CONCLUSION NF-kappaB plays a key role in AR. The excessively activated NF-kappaB promotes the transcription of ICAM-1 mRNA. ICAM-1 relates to the pathogenesis and development of AR.
Adult ; Case-Control Studies ; Female ; Humans ; Intercellular Adhesion Molecule-1 ; metabolism ; Male ; NF-kappa B p50 Subunit ; metabolism ; Nasal Mucosa ; metabolism ; RNA, Messenger ; genetics ; Rhinitis ; metabolism ; pathology ; Transcription Factor RelA ; metabolism

<b>OBJECTIVEb>To study the expression characteristics and biological significance of nuclear factor NF-kappaB p50 and NF-kappaB p65 in osteoporosis development. Adult mice were ovariectomized as models of experimental osteoporosis. In this way, we can explore the molecular mechanism of osteoporosis and understand the significance during the morbility of osteoporosis.

<b>METHODSb>Four-month female BALB/c mice were randomly divided into ovariectomized group and sham-operated group. 12 weeks after surgery, the mice were sacrificed after the measurement of bone mineral density (BMD). The NF-kappaB level in bone tissue were determined by immunohistochemistry and in situ hybridization techniques.

<b>RESULTSb>Compared with the sham-operated group, bone density, rarefaction of trabecula and the number of osteoblast decreased in the ovariectomized group, while the number of osteoclast increased in the ovariectomized group. The immunohistochemistry showed that NF-kappaB expressed in both groups. It mainly localized in the cytoplasm of osteoblast, osteocytes and marrow stroma cell. The expression level in the ovariectomized group was higher than that in the sham-operated group and was negatively correlated with the BMD (P < 0.01 or P < 0.05). In situ hybridization demonstrated that the expression levels of NF-kappaB 50mRNA and NF-kappaB 65mRNA in ovariectomized group were significantly higher than that of the sham-operated group (P < 0.05).

<b>CONCLUSIONb>The expression level of NF-kappaB significantly increased in the bone tissue of ovariectomized mice, and its abnormal expression was significantly correlated with BMD.

Animals ; Bone Density ; Bone and Bones ; metabolism ; Disease Models, Animal ; Female ; Immunohistochemistry ; In Situ Hybridization ; Mice ; Mice, Inbred BALB C ; NF-kappa B p50 Subunit ; biosynthesis ; genetics ; Osteoporosis ; etiology ; metabolism ; physiopathology ; Ovariectomy ; RNA, Messenger ; genetics ; Transcription Factor RelA ; biosynthesis ; genetics

Objective: Hyperbaric oxygen treatment (HBOT) has demonstrated efficacy in improving hearing levels of patients with idiopathic sudden sensorineural hearing loss (ISSHL); however, the underlying mechanisms are not well understood. HBOT alleviates the inflammatory response, which is mediated by Toll-like receptor (TLR) 4 and nuclear factor (NF)-κB. In this study we investigated whether HBOT attenuates inflammation in ISHHL patients alteration of TLR4 and NF-κB expression. Methods: ISHHL patients ( = 120) and healthy control subjects ( = 20) were enrolled in this study. Patients were randomly divided into medicine group treated with medicine only ( = 60) and HBO group receiving both HBOT and medicine ( = 60). Audiometric testing was performed pre- and post-treatment. TLR4, NF-кB, and TNF-α expression in peripheral blood of ISSHL patients and healthy control subjects was assessed by ELISA before and after treatment. Results: TLR4, NF-κB, and TNF-α levels were upregulated in ISSHL patients relative to healthy control subjects; the levels were decreased following treatment and were lower in the HBO group than that in the medicine group post-treatment ( < 0.05 and < 0.01). Conclusion HBOT alleviates hearing loss in ISSHL patients by suppressing the inflammatory response induced by TLR4 and NF-κB signaling.
Adolescent ; Adult ; Aged ; China ; Female ; Hearing Loss, Sensorineural ; therapy ; Hearing Loss, Sudden ; therapy ; Humans ; Hyperbaric Oxygenation ; Inflammation ; genetics ; therapy ; Male ; Middle Aged ; NF-kappa B p50 Subunit ; genetics ; metabolism ; Toll-Like Receptor 4 ; genetics ; metabolism ; Young Adult

We have observed earlier that testosterone at physiological concentrations can stimulate tissue factor pathway inhibitor (TFPI) gene expression through the androgen receptor in endothelial cells. This study further investigated the impact of testosterone on TFPI levels in response to inflammatory cytokine tumor necrosis factor-alpha (TNF-alpha). Cultured human umbilical vein endothelial cells were incubated in the presence or absence of testosterone or TNF-alpha. TFPI protein and mRNA levels were assessed by enzyme-linked immunosorbent assay and quantitative real-time reverse transcription polymerase chain reaction. To study the cellular mechanism of testosterone's action, nuclear factor-kappa B (NF-kappaB) translocation was confirmed by electrophoretic mobility shift assays. We found that after NF-kappaB was activated by TNF-alpha, TFPI protein levels declined significantly by 37.3% compared with controls (P < 0.001), and the mRNA levels of TFPI also decreased greatly (P < 0.001). A concentration of 30 nmol L(-1) testosterone increased the secretion of TFPI compared with the TNF-alpha-treated group. NF-kappaB DNA-binding activity was significantly suppressed by testosterone (P < 0.05). This suggests that physiological testosterone concentrations may exert their antithrombotic effects on TFPI expression during inflammation by downregulating NF-kappaB activity.
Androgens ; pharmacology ; Cells, Cultured ; Down-Regulation ; drug effects ; Drug Combinations ; Endothelium, Vascular ; drug effects ; metabolism ; Humans ; Infant, Newborn ; Lipoproteins ; genetics ; metabolism ; NF-kappa B p50 Subunit ; antagonists & inhibitors ; genetics ; RNA, Messenger ; metabolism ; Testosterone ; pharmacology ; Tumor Necrosis Factor-alpha ; pharmacology