This study examined the effects of Bangdeyun on the expressions of nuclear factor-kappaB (NF-kappaB), interferon-gamma (IFN-gamma) and interleukin-10 (IL-10) in the endometrium of mice with embryo implantation dysfunction (EID) during the implantation time (namely on pregnancy day 5, 6, 7 and 8) and explored the local immune regulatory effects of Bangdeyun. The gestational mice were randomly divided into normal group, model group and Bangdeyun-treated group. EID models of mice were established by using indomethacin. The endometrial expression of NF-kappaB was detected by immunohistochemistry and Western blotting. IFN-gamma and IL-10 were measured by enzyme-linked immunosorbent assay (ELISA). The results showed that in the normal group, NF-kappaB and IFN-gamma were weakly expressed and IL-10 was strongly expressed in the endometrium during the whole implantation period. In the model group, the expressions of NF-kappaB and IFN-gamma were increased on pregnancy day 5, 6 and 7, and IL-10 expression decreased during the whole implantation time when compared with those in the normal group (P<0.01 for all). In the Bangdeyun-treated group, little amount of NF-kappaB and IFN-gamma was expressed and IL-10 expression was strong, much the way they were expressed in the normal group (P>0.05). The expressions of NF-kappaB and IFN-gamma were much lower in the Bangdeyun-treated group than those in the model group on pregnancy day 5, 6 and 7 (P<0.01 for all), while the expression of IL-10 was much higher than in the model group during the whole implantation time (P<0.01). It was suggested Bangderun may favor a shift from Th1- to Th2-type immune response, therefore inhibiting the maternal immune rejection, inducing the immune tolerance and improving the fetal implantation.
Drugs, Chinese Herbal/*pharmacology ; Embryo Implantation, Delayed/*drug effects ; Embryo Implantation, Delayed/immunology ; Endometrium/*immunology ; Endometrium/metabolism ; Interferon-gamma/genetics ; Interferon-gamma/metabolism ; Interleukin-10/genetics ; Interleukin-10/metabolism ; NF-kappa B/genetics ; NF-kappa B/metabolism

OBJECTIVE: To explore the effects of advanced glycation end products (AGEs) on the activity of NF-kappaB and fibronectin (Fn) synthesis in the endothelial cells in aged rats. METHODS: Endothelial cells were cultured in M199 from the aorta of 24 month old rats and divided into 3 groups: Group A (5 mmol/L glucose) as controls, Group B (25 mg/L AGEs for 48 h), and Group C (50 mg/L AGEs for 48 h). The activity of NF-kappaB was evaluated by immunofluorescence and the expression of Fn mRNA was measured by reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: Compared with the controls, AGEs induced the activity of NF-kappaB and increased the Fn mRNA expression in a concentration-dependent manner (P<0.05). CONCLUSION The activity of NF-kappaB and up-regulated expression of Fn mRNA induced by AGEs may contribute to chronic complications of diabetes.
Animals ; Cells, Cultured ; Endothelial Cells ; drug effects ; metabolism ; Fibronectins ; genetics ; metabolism ; Glycation End Products, Advanced ; pharmacology ; NF-kappa B ; genetics ; metabolism ; RNA, Messenger ; genetics ; Rats ; Rats, Sprague-Dawley

OBJECTIVE: To study the role of Novaferon on TNF-α production and expression of NF-κB mRNA in monocytes isolated from normal human peripheral blood and to provide theoretical basis for treatment of immunological diseases with Novaferon. METHODS: Monocytes were isolated from the peripheral blood in 30 healthy volunteers and divided into 5 groups: group A was blank control, group B was stimulated by LPS without Novaferon intervention, group C by LPS together with Novaferon intervention, group D by LPS before Novaferon intervention, which group E by LPS after Novaferon intervention. We detected the concentration of TNF-α after LPS stimulation and Novaferon intervention in the supernatant by ELISA and expression of NF-κB mRNA by RT-PCR. RESULTS: Novaferon inhibited TNF-α production by monocytes isolated from healthy volunteers induced by LPS in vitro in group D compared with group B [(1446.76±72.36) pg/mL vs (946.46±46.12) pg/mL, P<0.01], and the rate was 29.7%. There was no significant change in TNF-α concentration in group C and E compared with group B [(1446.76±72.36) pg/mL vs (1275.62±87.75) pg/mL, P>0.05; (1446.76±72.36) pg/mL vs (1383.62±86.96) pg/mL, P>0.05]. There was significant change in NF-κB mRNA expression in group D compared with group B (0.2829±0.0365 vs 0.4994±0.0604, P<0.01). There was no significant change in NF-κB mRNA expression in group C and group E compared with group B (0.4716±0.0616 vs 0.4994±0.0604, P>0.05; 0.4767±0.0600 vs 0.4994±0.0604, P>0.05). CONCLUSION Novaferon can suppress TNF-α secretion by monocytes induced by LPS in vitro, and it can affect the immunity function of monocytes, which may be associated with the downregulation of NF-κB mRNA expression in monocytes.
Humans ; Lipopolysaccharides ; pharmacology ; Monocytes ; drug effects ; metabolism ; NF-kappa B ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Recombinant Proteins ; pharmacology ; Tumor Necrosis Factor-alpha ; antagonists & inhibitors ; metabolism

<b>OBJECTIVEb>To observe the effect of Shenluotong Decoction (SD) on serum levels of aldosterone, monocyte chemoattractant protein-1 (MCP-1), α-smooth muscle protein (α-SMA), and nuclear factor-KB (NF-&kappa;B) in obstructive nephropathy rats, and to explore the initial mechanism of SD for inhibiting renal interstitial fibrosis.

<b>METHODSb>Totally 48 healthy Wistar rats were randomly divided into the sham-operation group (n =12) and the model group (n =36). Renal interstitial fibrosis rat model was established by unilateral ureteral obstruction (UUO). After successful modeling, 36 rats were randomly divided into the model group, the Chinese medicine group, and the Western medicine group, 12 in each group. Eplerenone was added in the forage at the daily dose of 100 mg/kg for rats in the Western medicine group. Chinese medicine was added in the forage at the daily dose of 26 g/kg for rats in the Chinese medicine group. Equal volume of normal saline was administered to rats in the sham-operation group and the model group. All medication was performed once daily. The obstructive kidneys were extracted ten days after medication. The pathomorphological changes were observed. The contents of serum aldosterone and MCP-1, and the protein or mRNA expression of MCP-1, α-SMA, and NF-KB were detected.

<b>RESULTSb>Compared with the sham-operation group, infiltration of a large amount of inflammatory cells and collagen deposition significantly increased, serum contents of aldosterone and MCP-1 obviously increased (P < 0.01), the expression of MCP-1 mRNA and protein were significantly up-regulated (P <0.01), the protein expression of α-SMA and NF-KB were significantly enhanced in the model group (P <0.01). Com- pared with the model group, infiltration of inflammatory cells and renal collagen deposition were attenua- ted in the Chinese medicine group and the Western medicine group, the serum MCP-1 level were reduced, and the mRNA and protein expression of MCP-1 were significantly down-regulated (P <0.01), the protein expression of α-SMA and NF-KB were obviously inhibited (P <0. 01). At the same time, serum aldosterone level was reduced in the Chinese medicine group (P <0.01).

<b>CONCLUSIONSb>inflammatory lesions of the renal tissue could promote the progress of interstitial fibrosis in rats with obstructive nephropathy. SD could attenuate interstitial fibrosis through reducing serum contents of aldosterone and MCP-1, down-regulating MCP-1/ NF-KB, and inhibiting the expression of α-SMA.

Animals ; Chemokine CCL2 ; drug effects ; metabolism ; Down-Regulation ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Fibrosis ; Kidney ; drug effects ; Kidney Diseases ; drug therapy ; genetics ; NF-kappa B ; drug effects ; metabolism ; RNA, Messenger ; biosynthesis ; Rats, Sprague-Dawley ; Ureteral Obstruction ; drug therapy ; genetics

The migration of vascular smooth muscle cells (VSMCs) into the intima, an important step in injury-induced neointimal hyperplasia, requires the activation of nuclear factor-kappaB (NF-kappaB) and the consequent up-regulation of matrix metalloproteinase-9 (MMP-9). This study was undertaken to test for a possible effect of alpha-lipoic acid (ALA), a potent inhibitor of NF-kappaB, on MMP-9 expression. ALA inhibited high-glucose- and TNF-alpha-stimulated VSMC migrations in vitro. It also inhibited high-glucose- and TNF-alpha-induced increases in MMP-9 expression. The activity of MMP-9-promoter constructs with mutations in the NF-kappaB binding site was not inhibited by ALA, indicating an involvement of the NF-kappaB signaling pathway in the ALA-specific inhibition of MMP-9. These data suggest the possibility that ALA may be useful for the prevention of neointimal hyperplasia after angioplasty, by inhibiting the NF-kappaB/ MMP-9 pathway, especially with hyperglycemia.
Thioctic Acid/*pharmacology ; Rats, Sprague-Dawley ; Rats ; Promoter Regions (Genetics)/genetics ; NF-kappa B/*metabolism ; Muscle, Smooth, Vascular/cytology/drug effects/metabolism ; Matrix Metalloproteinase 9/genetics/*metabolism ; Male ; Gene Expression/*drug effects/*genetics ; Cells, Cultured ; Cell Movement/drug effects ; Animals

AIM: To investigate whether diosgenin could modulate tissue factor (TF) procoagulation activity, expression, and related signal transduction pathways. METHODS: Human THP-1 monocytic cells were exposed to tumor necrosis factor-α (TNF-α, 10 ng·mL(-1)) with or without diosgenin (0.01, 0.1, and 1 μmol · L(-1)) for 2 h or 5 h to induce TF procoagulant activity and expression, which were determined by the simplified chromogenic assay, reverse transcription-polymerase chain reaction (RT-PCR), real-time quantitative PCR, and Western blotting assays. In addition, the activation of the NF-κB, Akt, and MAPK signaling pathways were also measured by Western blotting. RESULTS: Diosgenin significantly inhibited TNF-α-induced TF procoagulant activity at concentrations of 0.01 to 1 μmol · L(-1) with IC50 of 0.25 μmol · L(-1). It also reduced protein expression and mRNA accumulation of TF dose-dependently in activated THP-1 cells. TNF-α stimulated significantly phosphorylation on Ser536 of NF-κB/p65, Ser473 of Akt at 5-15 min, and activations of IKK-β and ERK at 15-30 min. Diosgenin (1 μmol · L(-1)) could inhibit the phosphorylation of NF-κB/p65, IKK-β, Akt, ERK, and JNK, but had no remarkable effects on IκB and p38 phosphorylation in THP-1 cells. CONCLUSION Diosgenin inhibits TNF-α-induced TF activity and expression in monocytes, partly due to its down-regulation of the phosphorylation of NF-κB/p65, IKK-β, Akt, ERK, and JNK.
Diosgenin ; pharmacology ; Down-Regulation ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Humans ; MAP Kinase Signaling System ; drug effects ; Monocytes ; drug effects ; metabolism ; NF-kappa B ; genetics ; metabolism ; Proto-Oncogene Proteins c-akt ; genetics ; metabolism ; Tumor Necrosis Factor-alpha ; genetics ; metabolism

<b>OBJECTIVEb>To study the molecular mechanism of cyclooxygenase-2 (COX-2), one of effective ingredient of brucine, in inducing non-small cell lung cancer cell apoptosis.

<b>METHODb>COX-2 promoter, transcription factor deletion mutants and COX-2 mRNA 3'-UTR-containing report plasmids were transfected with Renillia to non-small cell lung cancer A549 cell, in order to detect the activity of report gene luciferase and minimum cis-acting element of COX-2 promoter inhibited by brucine. The influence of brucine on IkappaB phosphorylation and the nuclear translocation of p65 were detected by immunoblotting assay.

<b>RESULTb>Brucine significantly suppressed LPS-induced COX-2 promoter activation, but revealed minor impact on COX-2 mRNA stability. NF-kappaB in the vicinity of COX-2 promoter-262 was an important cis-acting element of brucine for inhibiting the activity of COX-2 promoter. Brucine was found to inhibit the phosphorylation of IkappaBalpha as well as the nuclear translocation of p65.

<b>CONCLUSIONb>Brucine can improve A549 cells apoptosis by inhibiting the activity of NF-kappaB and the subsequent COX-2 gene expression.

Biological Transport ; drug effects ; Carcinoma, Non-Small-Cell Lung ; genetics ; metabolism ; Cell Line, Tumor ; Cyclooxygenase 2 ; genetics ; Humans ; NF-kappa B ; metabolism ; Phosphorylation ; drug effects ; Promoter Regions, Genetic ; drug effects ; genetics ; RNA Stability ; drug effects ; Strychnine ; analogs & derivatives ; pharmacology

<b>OBJECTIVEb>To investigate the effects of emodin on expression of cytokines induced by lipopolysaccharide (LPS) in cultured human corneal fibroblasts in vitro.

<b>METHODSb>Primary human corneal fibroblasts of passages 4 were used in this research. Cells were treated with 10 microg/L LPS for 1, 2, 4, or 8 hours, which were pretreated with or without emodin for 30 minutes before LPS challenge. The degeneration of inhibitor of kappaB-alpha (I kappaB-alpha) and the effect of emodin on it were analyzed by Western blot analysis with a specific antibody. The cellular abundance of the mRNA of interleukin (IL)-6 and IL-8 from corneal fibroblasts under different conditions was determined by reverse transcriptase polymerase chain reaction (RT-PCR).

<b>RESULTSb>Compared with cells without LPS treatment, I kappaB-alpha level significantly decreased in every time point after LPS challenge (P < 0.01). Emodin inhibited the LPS-induced degeneration of I kappaB-alpha by corneal fibroblasts in a dose-dependent manner (P < 0.05). Compared with cells without LPS treatment, the expressions of IL-6 and IL-8 mRNA significantly increased in every time point after LPS challenge (P < 0.01). At the same time, the expressions of the mRNA of IL-6 and IL-8 induced by LPS in corneal fibroblasts were also inhibited by emodin in a dose-dependent manner (P < 0.05).

<b>CONCLUSIONb>Emodin can inhibit the expressions of IL-6 and IL-8 mRNA induced by LPS in corneal fibroblasts, which maybe via inhibiting the degeneration of I kappaB-alpha and suppressing the activation of nuclear factor-kappaB.

Cells, Cultured ; Cornea ; cytology ; drug effects ; metabolism ; Drug Antagonism ; Emodin ; pharmacology ; Fibroblasts ; drug effects ; metabolism ; Humans ; Interleukin-6 ; genetics ; metabolism ; Interleukin-8 ; genetics ; metabolism ; Lipopolysaccharides ; toxicity ; NF-kappa B ; metabolism ; RNA, Messenger ; genetics

<b>OBJECTIVEb>To study whether the protective effects of glutamine against intestinal injury are associated with the regulation of intestinal secretion of NF-&kappa;B and TNF-α by glutamine.

<b>METHODSb>Twenty-four 10-day-old Wistar rat pups were randomly intraperitoneally injected with normal sodium of 1 mL/kg (control group; n=8), LPS of 5 mg/kg (endotoxemia group; n=8) or LPS of 5 mg/kg plus glutamine of 10 mL/kg (glutamine group; n=8). The rats were sacrificed 3 hrs after injection. A segment of distal ileum was dissected. The pathologic changes of the small intestine were observed under an optical microscope (hematoxylin-eosin staining). The expression of intestinal NF-&kappa;B mRNA and protein were detected by RT-PCR and Western blot respectively. The TNF-α level in intestinal tissues was measured using ELISA.

<b>RESULTSb>The structure of the small intestine in the control group remained normal. The inflammation cells infiltration and the edema of interstitial substance and epithelials were observed in the endotoxemia and glutamine groups. The pathologic changes in the glutamine group were significantly alleviated when compared with the endotoxemia group. The expression of NF-&kappa;B mRNA and protein in the glutamine group was significantly lower than that in the endotoxemia group (P<0.01). The secretion of TNF-α in the glutamine group was significantly lower than that in the endotoxemia group (P<0.01).

<b>CONCLUSIONSb>The protective effects of glutamine against intestinal injury may be associated with the down-regulation of intestinal NF-&kappa;B mRNA and protein expression and the reduction of the secretion of TNF-α.

Animals ; Endotoxemia ; metabolism ; Female ; Glutamine ; pharmacology ; Intestines ; drug effects ; metabolism ; Male ; NF-kappa B ; analysis ; genetics ; secretion ; Rats ; Rats, Wistar ; Tumor Necrosis Factor-alpha ; secretion

Reactive oxygen species (ROS) has been implicated as an inducer of NF-kappaB activity in numbers of cell types where exposure of cells to ROS such as H2O2 leads to NF-kappaB activation. In contrast, exposure to oxidative stress in certain cell types induced reduction of tumor necrosis factor (TNF)-induced NF-kappaB activation. And various thiol-modifying agents including gold compounds and cyclopentenone prostaglandins inhibit NF-kappaB activation by blocking IkappaB kinase (IKK). To understand such conflicting effect of oxidative stress on NF-kappaB activation, HeLa cells were incubated with H2O2 or diamide and TNF-induced expression of NF-kappaB reporter gene was measured. NF-kappaB activation was significantly blocked by these oxidizing agents, and the inhibition was accompanied with reduced nuclear NF-kappaB and inappropriate cytosolic IkappaB degradation. H2O2 and diamide also inhibited IKK activation in HeLa and RAW 264.7 cells stimulated with TNF and lipopolysaccharide, respectively, and directly blocked IKK activity in vitro. In cells treated with H2O2 alone, nuclear NF-kappaB was induced after 2 h without detectible degradation of cytosolic IkBa or activation of IKK. Our results suggest that ROS has a dual effect on NF-kappaB activation in the same HeLa cells: it inhibits acute IKK-mediated NF-kappaB activation induced by inflammatory signals, while longer-term exposure to ROS induces NF-kappaB activity through an IKK-independent pathway.
Cell Nucleus/drug effects/metabolism ; Cytosol/drug effects/metabolism ; Diamide/pharmacology ; Hela Cells/drug effects/metabolism ; Human ; Hydrogen Peroxide/pharmacology ; I-kappa B/drug effects/metabolism ; NF-kappa B/drug effects/genetics/*metabolism ; Oxidants/pharmacology ; *Oxidative Stress ; Protein-Serine-Threonine Kinases/metabolism ; Signal Transduction/drug effects ; Time Factors ; Transcription, Genetic ; Tumor Necrosis Factor/pharmacology