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@#To investigate the therapeutic effect and mechanism of sodium formononetin-3′-sulphonate (SFS) on collagen-induced rheumatoid arthritis in mice, C57 mice were induced with chicken type II collagen to establish a model of rheumatoid arthritis (collagen-induced arthritis, CIA), and were injected intraperitoneally with different doses of SFS (50,100,200 mg/kg). Body weight, food intake and foot swelling of all groups were observed during the experiment.After the treatment, TNF-α, IL-6, and IL-10 in the serum were detected with the CBA kit; NF-κB p65, p-NF-κB p65 (p-p65), TIPE2, PCNP and IκB-α in spleen tissue were determined by Western blot; the organ index, pathological changes of ankle joint cartilage tissue and the positive expression of NF-κB p65 in ankle joint tissue were also observed.The results showed that, compared with the model group, the body weight and food intake of mice in the treatment group increased, while the degree of foot swelling decreased; the expression levels of inflammatory factors TNF-α and IL-6 in serum decreased, while the expression of anti-inflammatory factor IL-10 increased; the expression levels of NF-κB p65, p-p65 and PCNP in spleen tissue decreased, while the expression of TIPE2 and IκB-α protein increased; the index of spleen and thymus of the CIA mice in the treatment group, the infiltration of inflammatory cells in the ankle joint, the destruction of synovial tissue and cartilage, and the positive expression of NF-κB p65 decreased.Among them, the high-dose group of SFS showed a better therapeutic effect.It is suggested that SFS has a therapeutic effect on CIA mice, and the mechanism may be achieved by regulating the NF-κB p65 signaling pathway and inhibiting the expression of inflammatory factors.
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Objective:To investigate the effect of Xiao Xumingtang combined with super-acupuncture along governor meridian on autophagy-related protein nuclear factor-kappa B p65 (NF-κB p65) in cerebral ischemia-reperfusion model rats, so as to study the relationship between autophagy-related protein NF-κB p65 and brain protection mechanism, and look for the best intervention time point of acupuncture. Method:A total of 152 adult SD male rats were randomly divided into sham operation group, model group, high-dose Xiao Xumingtang group (high-dose drug group), low-dose Xiao Xumingtang group (low-dose drug group) and acupuncture group. There were seven groups including high-dose Xiao Xumingtang + acupuncture group (high acupuncture group) and low-dose Xiao Xumingtang + acupuncture (low acupuncture group). Model group, high-dose drug group, low-dose drug group, and acupuncture group were divided into 4 subgroups according to 30 minutes, 2, 4， 6 h of ischemia-reperfusion, with 6 animals in each group. After successful modeling, according to Zea Longa's neural function score, eligible rats were included into the corresponding groups. The sham operation group only received carotid artery dissection, the model group was only modeled without any treatment, high and low-dose Xiao Xumingtang groups were calculated based on the body surface area of the animal and given 60 g·kg-1·d-1 and 15 g·kg-1·d-1 drug by gavage for treatment, acupuncture was performed to smooth governor meridian and regulate the mind. After 14 days of consecutive treatment, neurological function was scored. Western blot was used to detect the expression of autophagy-related protein NF-κB p65 in rat brain tissue. Result:Compared with the sham operation group, the neurological impairment scores of the model group, the high-dose drug group, the low-dose drug group, the acupuncture group, the high acupuncture group, and the low acupuncture group were significantly increased at each time point (P<0.01). The neurological impairment scores were significantly lower at each time point than those of the high-dose drug group, low-dose drug group, acupuncture group, high acupuncture group, and low acupuncture group (P<0.01), compared with the sham operation group, NF-κB p65 in model group, high-dose drug group, low-dose group, acupuncture group, high acupuncture group and low acupuncture group was significantly increased in the brain tissue at each time point (P<0.01), compared with the model group, the expression of NF-κB p65 protein in the brain tissue of model group, high-dose drug group, low-dose group, acupuncture group, high acupuncture group and low acupuncture group was decreased at each time point (P<0.05), particular in the high acupuncture group (P<0.01). Conclusion:Xiao Xumingtang combined with ultra-early acupuncture along governor meridian can significantly alleviate neurological impairment in rats with cerebral ischemia-reperfusion model. Xiao Xumingtang combined with ultra-early acupuncture along governor meridian can inhibit cerebral ischemia-reperfusion model rats. The activity of autophagy-related protein NF-κB P65 protects the brain function. There is no significant difference in the brain protective effect of Xiao Xumingtang combined with ultra-early acupuncture along governor meridian within 6 hours.
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Objective@#To study the effects of Phellodendron amurense on herpes simplex virus 1 (HSV-1) and cytokines, and to explore the mechanism of Phellodendron amurense inhibiting HSV-1 virus through multiple channels.@*Methods@#Viruses were inoculated into medicine treated HeLa cells. The proliferation of virus was observed by fluorescence microscopy. The transcriptional levels of glycoprotein gD and functional protein US1 on the surface of virus envelope were detected by quantitative (q) PCR. After incubating HeLa cells for 24 h, qPCR was used to detect the expression of intrinsic immune factors such as IP-10, IL-12, IFN-gamma and transcription factor NF-kappa B (P65). The expression and nuclear location of NF-kappa B (P65) protein were detected by immunofluorescence.@*Results@#Fluorescence showed that the proliferation of virus decreased significantly at 8 and 40 ng/ml (P<0.01), and the transcription levels of viral protein gD and US1 decreased (P<0.05). After incubation for 24 hours, the transcription levels of IP-10, IL-12 and IFN-gamma in HeLa cells increased significantly (P<0.01). The transcription level of transcription factor NF-kappa B (P65) also increased (P<0.05), and immunofluorescence showed that the nuclear penetration rate of p65 subunit of NF-kappa B (P65) in each group increased (P<0.05).@*Conclusions@#Phellodendron amurense extract can inhibit HSV-1 by inhibiting the transcription of viral functional protein and promoting the expression of cellular immune factors.
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Objective: To study the effect of modified Erchentang on expressions of Toll-like receptor 4 (TLR4), myeloid differentiation factor (MyD88) and nuclear factor-κB (NF-κB) genes in the lung tissue homogenate of rats with chronic obstructive pulmonary disease (COPD). Method: Forty SD rats were randomly divided into normal group, model group, modified Erchentang group and EVP4593 (NF-κB inhibitor) group. Rat COPD models were prepared through cigarette smoke and tracheal dripping with lipopolysaccharide (LPS). After the modeling, normal and model groups were intragastrically given normal saline solution, EVP4593 group was given EVP4593(1 mg · kg-1) through subcutaneous injection, and modified Erchentang group was given corresponding herbal drugs intragastrically (10 g · kg-1) for 14 days. The levels of high mobility group box 1(HMGB1), chemokines CXCL-2, CXCL-3 and monocyte chemoattractant protein-1 (MCP-1) in rats serum were detected by enzyme-linked immunosorbent assay in rats serum. The expressions of Toll-like receptors 4(TLR4), myeloid differentiation factor (MyD88) and nuclear factor-κB p65 (NF-κB p65) mRNA were detected by Real-time fluorescence quantitative PCR (Real-time PCR) method. Western blot were used to detect the levels of TLR4, MyD88, NF-κB p65 and p-NF-κB p65 protein. Immunohistochemistry (IHC) method was used to detect the localization and expressions of TLR4, MyD88 and p-NF-κB p65 protein in the lung tissue. Result: The mRNA and protein expressions of TLR4, MyD88 and NF-κB p65 were increased significantly (PPκB p65 mRNA and protein were decreased significantly (PConclusion: Modified Erchentang may inhibit the inflammatory response of COPD effectively. The mechanism may be related to the inhibition of the expressions of the signal molecule genes involved in the TLR4/MyD88/NF-κB pathway and the reduction of the release of HMGB1, CXCL-2, CXCL-3 and MCP-1.
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[Objective]Examine the expression of microsomal prostaglandin E synthase-1(mPGES-1)and nuclear transcription factor kappa B p65(NF-κB p65)in diffuse large B cell lymphoma(DLBCL),assessing their correlation with clinical variables,prognosis and potential clinical valve.[Methods]The immunohistochemistry was uesd to investigate the expression of mPGES-1 and NF-κB p65 in 83 DLBCL patiens'tissues.The relationship between these two proteins and the clinical variables and prognosis of these patients was evaluated.[Results]The high expression of mPGES-1 and NF-κB p65 were observed in 65.1%(54/83)and 73.5%(61/83)cases of DLBCL,respectively.The expression level of NF-κB p65 was positively correlated with mPGES-1 expression(P<0.05).The expression of these two proteins was found to be significantly associated with B cell lymphoma/leukemia-2 protein(BCL-2),higher expression of Ki67,higher lactate dehydrogenase (LDH),more extranodal lesions,advanced Ann Arbor stage and higher international prognostic index(IPI)score(P<0.05). In addition,NF-κB p65 was related with multiple myeloma oncogene 1(MUM1),pathological type(P<0.05). Both mPGES-1 and NF-κB p65 overexpression was correlated with worse overall survival(OS)while NF-κB p65 was an in-dependent prognostic factor for OS of DLBCL(P<0.05).[Conclusions]mPGES-1 and NF-κB p65 were highly expressed in DLBCL and closely linked with each other. The overexpression of mPGES-1 and NF-κB p65 was correlated with tumor progression and unfavorable prognosis in patients with DLBCL.
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Objective To observe the effect of electroacupuncture(EA)at Baihui(GV20)and Siguan(Hegu/LI4 and Taichong/LR3)of affected side on expression of Tax1-binding protein 1(TAX1BP1)in cerebral cortex in focal cerebral ischemia-reperfusion rats,so as to in-vestigate its protective mechanism in inhibiting nuclear factor kappa B(NF-κB)signaling pathway and promoting neurobehavioral recovery. Methods A total of 105 Sprague-Dawley rats were randomly assigned into sham group,model group and EA group.Each group was ran-domly assigned into reperfusion six hours,twelve hours,24 hours,48 hours,72 hours groups after two hours of ischemia.The model was es-tablished by right middle cerebral artery occlusion and reperfusion.EA group received electroacupuncture at Baihui and left Siguan(Hegu and Taichong)acupoints.Neurobehavioral evaluation,TAX1BP1 protein expression,TAX1BP1 positive cell count,zink finger protein A20 expression, and nuclear NF-κB p65 protein expression were tested in each group. Results There was no neurological deficit in the sham group. Compared with the model group, the neurological scores at 48 hours, 72 hours after reperfusion decreased in EA group (P<0.05). Compared with the control group,the TAX1BP1 expression at twelve hours,24 hours and 48 hours after reperfusion increased in the model group(P<0.05),and further increased at twelve hours,24 hours,48 hours and 72 hours after reperfusion in EA group(P<0.05),and peaked at 24 hours after reperfusion.Compared with the sham group,the expression of A20 and NF-κB p65,and the number of TAX1BP1 positive cells increased in the model group(P<0.05),and the expression of A20 and the number of TAX1BP1 positive cells further increased,(P<0.05)and the expression of NF-κB p65 decreased in EA group(P<0.05)at 24 hours after reperfusion.Immunofluorescence labeling indicat-ed that TAX1BP1 protein primarily expressed in the cytoplasm,TAX1BP1 protein and A20 protein co-expressed in the cytoplasm.Immuno-histochemistry showed indicated that NF-κB p65 mainly expressed in the nucleus in the model groupr,and mainly expressed in the cyto-plasm in the EA group.Conclusion Electroacupuncture could significantly inhibit neuronal NF-κB signaling pathway and promote neurobe-havioral recovery in focal cerebral ischemia-reperfusion,which may related with up-regulating TAX1BP1 protein expression.
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Objective To investigate the effect of different doses of dexmedetomidine (DEX) on myocardial ischemia-reperfusion injury (MIRI) in rats.Methods Seventy-five SD male rats weighing between 250 and 300g were divided into sham group,MIRI group,small-dose DEX group,medium-dose DEX group and high-dose group according to the random number table,with 15 rats per group.Threading the left anterior descending coronary artery was done only in sham group,but the MIRI model was produced in the rest groups by ligation of the artery for 30 minutes followed by 120 minutes of reperfusion.Fifteen minutes before the ligation,small-,medium-and high-dose DEX groups were injected 2.5,5 and 10 μg · kg-1 · h-1 of DEX respectively until the end of reperfusion.Instead,an equal volume of normal saline was given in sham and MIRI groups.At the end of reperfusion,five rats in each group were used to determine the myocardial infarct size,and arterial blood samples and myocardial tissues from ten rats in each group were used to measure serum levels of interleukin-1β (IL-β) and serum tumor necrosis factor-α (TNF-α),expression of myocardial nuclear factor kappa B p65 (NF-κB p65) and change of myocardial pathomorphology.Results Myocardial infract size,degree of myocardial pathomorphology structure damage,serum levels of IL-1 β and TNF-αt and expression of myocardial NF-κB p65 in sham group were significantly lower in sham group than other groups (P ＜ 0.05).Above mentioned parameters in small-,medium-and high-dose DEX groups were all significantly decreased compared to MIRI group (P ＜ 0.05),and the decrease was most significant in medium-dose DEX group (P ＜ 0.05).Conclusions DEX can attenuate the MIRI in rats and the possible mechanism is suppressing the release of NF-κB p65,which can reduce serum pro-inflammatory cytokines like TNF-α and IL-1β.And mediumdose DEX exhibits better protective effect.
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Background:Hepatic veno-occlusive disease( HVOD) is a disease characterized by hepatomegaly,jaundice, ascites,weight gain and lack of effective treatment currently. Our prophase research showed that ligustrazine had therapeutic effect on Sedum aizoon induced HVOD in mice. Aims:To investigate the mechanism of therapeutic effect of ligustrazine on Sedum aizoon induced HVOD in mice. Methods:A total of 115 mice were randomly divided into 4 groups:mice in group A were intragastrically administrated with 30 mg·kg-1 ·d-1 Sedum aizoon to induce HVOD and served as model group;mice in group B were given 30 mg·kg-1 ·d-1 Sedum aizoon + 100 mg·kg-1 ·d-1 ligustrazine and served as low dose ligustrazine intervention group;mice in group C were given 30 mg·kg-1 ·d-1 Sedum aizoon + 200 mg·kg-1 ·d-1 ligustrazine and served as high dose ligustrazine intervention group;mice in group D were given 30 mg·kg-1 ·d-1 PBS and served as normal control group. After 30 days,all the mice were sacrificed. HE staining and Masson staining were performed for histological examination. The mRNA and protein expressions of tissue factor(TF),nuclear factor(NF)-κBp65 and early growth response factor( Egr)-1 in liver tissue were determined by RT-PCR and Western blotting, respectively. Results:HE staining and Masson staining histological examination showed that ligustrazine could obviously ameliorate the pathological injury of liver tissue in HVOD mice. Compared with group D,the mRNA and protein expressions of TF,NF-κBp65,Egr-1 were significantly increased in group A( P 0. 05). Conclusions:Ligustrazine has therapeutic effect on HVOD,the possible mechanism is that ligustrazine could interrupt the activation of coagulation system by reducing the expression of TF via down regulating the expressions of NF-κBp65 and Egr-1,especially in high dose ligustrazine group.
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Objective To study the effects of exogenous hydrogen sulfide (H2S) on myocardial fibrosis of diabetic mice and the mechanism thereof. Methods Twenty-four male adult C57BL/6J mice were randomly divided into 4 groups (6 each): normal control group (NC group), diabetes mellitus group (ALX group), diabetes mellitus treated with low dose NaHS group (ALX+LDNaHS group) and diabetes mellitus treated with high dose NaHS group (ALX+HDNaHS group). The diabetic mouse model was established by intraperitoneal injection of alloxan at 200mg/kg. Mice in NC group and ALX group drank tap water freely everyday, and in ALX+LDNaHS group and ALX+HDNaHS group were provided with sodium hydrosulfide (NaHS, donor of H2S, 30μmol/L and 90μmol/L, respectively) in drinking water, and the histological specimens of mice were examined 8 weeks later. The morphological changes of cardiac tissue were examined with HE staining. The expressions of mRNA of p38mitogen-activated protein kinase (p38MAPK) and nuclear transcription factor kappa B p65 (NF-κB p65) were detected by Real-time PCR, and the expressions of p-p38MAPK, p-NF-κB p65 and Collagen I proteins were detected by Western blotting. Results Compared with NC group, the expression of collagen increased and there was fibrillation in the myocardial matrix, the expressions of mRNA of p38MAPK and NF-κB p65 increased obviously (P<0.01), the protein expressions of p-p38MAPK, p-NF-κB p65 and Collagen I increased significantly (P<0.01) in ALX group; after intervention of H2S, the cardiac muscle fibers were parallel arranged, the expression of collagen was visibly decreased and there was less fibrillation in the myocardial matrix, the expressions of mRNA of p38MAPK and NF-κB p65 were obviously decreased (P<0.01), the protein expressions of p-p38MAPK, p-NF-κB p65 and Collagen I were significantly decreased (P<0.01), and more improvement was observed in ALX+HDNAHS group than in ALX+LDNAHS group (P<0.05). Conclusion H2S can ameliorate myocardial fibrosis in diabetic mice, which might be related to the regulation of p38MAPK and NF-κB p65-mediated inflammatory reaction.
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Objective To investigate the effects of HMG-CoA reductase inhibitor rosuvastatin on atherosclerosis and lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1), and nuclear factor (NF)-kB p65 expressions in apolipoprotein E (ApoE)-deficient mice. Methods Twenty six-week-old ApoE-deficient male mice were randomly divided into hyperlipidemia model group (n=10) and rosuvastatin group (n=10), and they were fed high-fat diet for 13 weeks. Ten six-week-old C57BL/6J (wild type, WT) male mice were selected as normal control group, and were fed normal diet for 13 weeks. After 13 weeks, blood was drawn from the mice to determine serum levels of total cholesterol (TCH), triglyceride (TG), and low density lipoprotein cholesterol (LDL-C). These mice were sacrificed, and their aortas were obtained and examined with HE staining; Western blotting and RT-PCR were used to analyze LOX-1, NF-kB p65 expression intensity in aorta tissue quantitatively. Results The serum level of TCH, TG and LDL-C in rosuvastatin group were lower than those in hyperlipidemia model group (P<0.05). Pathological observation showed that atherosclerotic lesions of the aortas were aggravated significantly in hyperlipidemia model group but alleviated in rosuvastatin group compared with normal control group. Compared with normal control group, LOX-1, NF-kB p65 protein and mRNA expressions significantly increased in hyperlipidemia model group (P<0.05) and reduced in rosuvastatin group (P<0.05). Conclusions Rosuvastatin may lower blood lipid significantly, alleviate the degree of atherosclerotic lesions, and inhibit LOX-1, NF-kB p65 protein and mRNA expressions in the aortic tissue of ApoE-deficient mice. Its anti-athrosclerosis effect is related to down regulation of LOX-1 and NF-kB p65 expressions.
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This study aimed at identifying the molecular mechanisms and effects of hypertonicity on mucin5AC (MUC5AC) expression in airway epithelial cells for which immortalized human bronchial epithelial (16HBE) cells were cultured in 600 mOsm/L hypertonic medium for different times in vitro. Proteins of MUC5AC and β-catenin, Cyclin D1, NF-κB p65 were detected by enzyme linked immunosorbent assay (ELISA), reverse transcription (RT)-PCR and western blotting assays. After transfection of β-catenin siRNA in 16HBE cells, the levels of β-catenin, Cyclin D1, NF-κB p65 and MUC5AC protein were detected. Results showed that the levels of mRNAs and proteins of target genes increased significantly after the exposure to hypertonic conditions and the expression content increased in a time-dependent manner. Furthermore, transfection with β-catenin siRNA attenuated the expression of these genes. These results suggested that the Wnt/β-catenin and NF-κB signaling pathways played essential roles in inducing the MUC5AC hypersecretion in 16HBE cells in response to hypertonicity.
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BACKGROUND/AIMS: Nuclear factor-kappa B p65 (NF-kappa B p65), nuclear factor-kappa B1 p50 (NF-kappa B p50) have been shown to play a role in cell proliferation, apoptosis, cytokine production, and oncogenesis. Recently, p38 mitogen-activated protein kinase (MAPK)/ NF-kappa B/ cyclin D1 signaling pathway has been shown to play an important part in the pathogenesis of human cancers. This study was designed to investigate the expression of NF-kappa B p65, NF-kappa B p50, p38 MAPK alpha, and cyclin D1 proteins in premalignant lesions of colon and colorectal adenocarcinoma. METHODS: Paraffin sections of 20 normal mucosa, 20 low-grade tubular adenoma, 20 high-grade tubular adenoma and 64 adenocarcinoma tissues were analysed immunohistochemically for the expression of NF-kappa B p65, NF-kappa B p50, p38 MAPK alpha, and cyclin D1 proteins. RESULTS: The expression of NF-kappa B p65, NF-kappa B p50, and p38 MAPK alpha proteins were significantly higher in adenocarcinoma tissue in comparison with that in normal mucosa, low-grade tubular adenoma, and high-grade tubular adenoma tissues. Expression of NF-kappa B p50 was more frequent in poorly differentiated histologic grade, presence of nodal metastasis, and advanced stage. Expression of p38 MAPK alpha protein was higher in advanced tumor stage, presence of nodal metastasis and advanced stage. Synchronous expression of NF-kappa B p65, NF-kappa B p50, p38 MAPK alpha, and cyclin D1 proteins were significantly higher in adenocarcinoma tissue. CONCULSIONS: With the increased expression of NF-kappa B p65, NF-kappa B p50, and p38 MAPK alpha proteins, p38 MAPK/ NF-kappa B/ cyclin D1 signaling pathway may play a role in the pathogenesis of colorectal carcinoma.
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Female , Humans , Male , Middle Aged , Adenocarcinoma/enzymology , Colorectal Neoplasms/enzymology , Cyclin D1/immunology , Data Interpretation, Statistical , Immunohistochemistry , Intestinal Mucosa/metabolism , NF-kappa B/immunology , NF-kappa B p50 Subunit/immunology , Neoplasm Staging , Precancerous Conditions/enzymology , Transcription Factor RelA/immunology , p38 Mitogen-Activated Protein Kinases/metabolismABSTRACT
Objective To study the effect of simvastatin(SV) on NF-kappa B p65 expression in liver of insulin resistant rats. Methods Male Wistar rats were divided into two groups:basic chow(NC,n=15)or high-fat diet(HF,n=20)for 10 weeks feeding, then assessed by euglycemic-hyperinsulinemia clamp technique. Rats in HF group were treated with(HFS,n=5)and without (HFN,n=5) SV 10mgkg-1d-1 in gavage for 5 weeks.NF-kappa B p65 expressions were tested with Western blot. Results The glucose infusion rate (GIR) in HF group decreased significantly compared with NC group (P
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Objective To explore the expression of PTEN,NF-KB p65 and Survivin protein and the influence in the genesis and development of endometrial carcinoma(EC).Methods The expression of PTEN.NF-KB and Survivin in 63 cases of EC and 20 cases normal endometrial tissue specimens were detected by immunohistochemistry.Resuits There were obvious differences among the positive rates of PTEN.N-KB p65 and Survivin protein in EC compared with in normal endometrial tissue specimens (P＜0.001).The clinicopathological characteristics of endometrial carcinoma with PTEN,NF-KB p65 and Survivin were as follows:the expression of PTEN gene was positively correlated with the degree of histological differentiation(P＜0.001),and it was negatively correlated with lymph node metastasis and TNM (P＜0.005).There was inverse correlation between the expression of NF-KB p65 gene and the degree of histological differentiation(P＜0.05).It was positively correlated with lymph node metastasis,deep myometrial invasion and TNM in EC (P＜0.05).The expression of Survivin was positively correlated with the clinical stage and lymph node metastasis and deep myometrial invasion in EC(P＜0.05).Conclusion There are difierent extent action of PTEN.NF-KB p65 and Survivin significantly in genesis and development of EC.Detection of PrEN combined with NF-KB p65 and Survivin is valuable for early diagnosing and evaluating malignancy extent of EC.
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Objective:To investigate the expression of NF?B p65 and PTEN in laryngeal squamous cell carcinoma(LSCC) and their clinical significance.Methods:Immunohistochemical staining was used to examine the expression of NF?B p65 and PTEN in 52 cases of LSCC and 30 cases of adjacent normal tissues.Results:The expression positive rates of NF?B p65 in LSCC were obviously higher than those in adjacent normal tissues(P0.05).The Spearman analysis indicated that the expression level of NF?B p65 was negatively correlated with that of PTEN significantly(rs=-0.580,P
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Objective:To investigate the expression and significance of NF-?B,and ICAM-1 in laryngeal carcinoma patients.Methods:Two groups of laryngeal carcinoma tissue (one group derived from the laryngeal carcinoma patients;the other from the normal persons) were detected for NF-kappa B P65 protein and ICAM-1′s expressions by immunohistochemistry staining.Results:(1)The expressions of NF-kappa B P65 protein and ICAM-1 of the laryngeal carcinoma patients were much more strengthened than those of the normal persons(P
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Objective:To investigate the in vitro effect of Bifidobacterium BB12 on IL-8 secretion by Caco-2 cells induced with S.enteritidis.Methods:The coculture systems of Caco-2 cells with S.enteritidis alone or combined with the strain of Bifidobacterium BB12 were established. The expression of IL-8 mRNA and NF-?B p65 in the cells were examined by reverse transcription PCR, real-time PCR and EMSA, and IL-8 in the supernatant was measured by ELISA.Results:In control group, there was little IL-8 mRNA expression in Caco-2 cells, NF-?B p65 was seldom in Caco-2 cells’ nuclei, and the concentration of IL-8 in the supernatant was as few as 126 ng/L. After stimulated by the S.enteritidis, the expression of IL-8 mRNA and activated NF-?B p65 was found in Caco-2 cells (P
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AIM:To investigate the expressions of BAG-1 and NF-?B P65 in the multiple myeloma(MM)and approach the effects of BAG-1 and NF-?B P65 in the pathogenesis of MM and their clinical significance,which would be useful to find a theoretical basis for the treatment of MM.METHODS:The patients diagnosed MM in Yijishan hospital of wannan medical college from 2006.10 to 2008.10 as the experimental group,while the bone trauma patients selected as the control group.The mononuclearcells of bone marrow in the patients were extracted and the BAG-1and NF-?B P65 mRNA RT-PCR were detected.RESULTS:The expression of BAG-1 mRNA(71.4%)in the MM patients was higher than that in the control group(P=0.007).The expression of NF-?B P65 mRNA(75.0%)in the MM patients was higher than that in the control group(P=0.004).There was a positive correlation between the BAG-1 and NF-?B P65 mRNA.The tumor load was high in the MM patients with overexpression of BAG-1 and NF-?B P65,which was ralated to a number of clinical parameters indexes.BAG-1 and NF-?B P65 were positive expressed of MM tumor load,relatived with a number of clinical indicators.CONCLUSION:The expressions of BAG-1 and NF-?B P65 in MM are high,there are no difference between the experimental group and control group.BAG-1 and NF-?B P65 were related to a number of clinical indexes.BAG-1 and NF-?B P65 may be involved in the occurrence and the development of disease,and they can be used as one of the prognosis indexes.
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Background and purpose:Anticancer mechanism of epigallocatechin-3-gallate(EGCG)remains unclear.This study was to investigate the effects of EGCG on induction of apoptosis by TNF-? through translocation of NF-?B/p65 into SGC-7901 cellular nucleus.Methods:Hoechst staining method was used to display the apoptosis form of SGC-7901 induced by EGCG.The disposition of NF-?B/p65 was detected by Western blot.The effect of EGCG on induction of apoptosis was analyzed by flow cytometry(FCM)and DNA agarose gel electrophoresis.Results:Hoechst staining method showed that EGCG could significantly induce apoptosis of SGC-7901.After being exposed to EGCG,the cell nucleus changed to shiver and pallor.After being treated by 10 ng/ml TNF-? at various times(30,60,90 and 120 min),an inductor of regulating NF-?B/p65,the expression of NF-?B/p65 in the cell nucleus increased obviously.It displayed a platform after being treated for 60 minutes.Incubation with 60 ?g/ml EGCG at various times(0,12,24,36,48 h)before TNF-?(10 ng/ml,60 min)treatment,the expression of NF-?B/p65 in the cell nucleus decreased in a time-depended manner that reached plateau after treated for 24 h.Being treated with EGCG for 24 h at various concentrations(40,60,80,100 ?g/ml)before the TNF-?(10 ng/ml,60 min),the expression of NF-?B/p65 in the cell nucleus decreased in a dose-depended manner.Analysis by flow cytometry showed that the percentages of apoptosis was 3.7% in cells exposed to 10 ng/ml TNF-?,60 min,apoptotic percentage was 16.6% after cells exposed to 60 ?g/ml EGCG(24 h)before TNF-?(10 ng/ml,60 min).It could reach to 21.4% after cells were exposed to PDTC(100 ?mol/L,30 min,before TNF-?),a suppressor of NF-?B.In DNA agarose gel electrophoresis,"Ladder"bands occurred after the treatment with EGCG(60 ?g/ml,24 h)before TNF-?(10 ng/ml,60 min).The "Ladder"bands also occurred after incubation with PDTC(100 ?mol/L,30 min)before TNF-?(10 ng/ml,60 min).Conclusions:EGCG could inhibit the translocation of NF-?B/p65's into the cell nucleus induced by TNF-?,and could significantly induce apoptosis through this pathway.
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Objective To explore the expression of silencer of death domains(SODD) and its clinical significance and relationship with phospho-NF-?B-p65 proteins in bone marrow cells of acute lymphoblastic leukemia(ALL)in children,and the expression of SODD and phospho-NF-?B-p65 in Jurkat cells treated with chemotherapeutic drugs in order to find a new chemotherapeutic target.Methods The expressions of SODD and phospho-NF-?B-p65 proteins in bone marrow cells were detected by immunohistochemistry in 25 children with ALL.The apoptosis incidence was measured by Annexin-V-Fluorescence/PI double-labeling flow cytometry and the expression of SODD and phospho-NF-?B-p65 proteins were determined by Western blotting in Jurkat cells.Results It was found that the expression of SODD and active p65 expression in ALL were significantly higher than those in healthy control group.The expression of SODD and phospho-NF-?B-p65 proteins in the high-risk(HR) group was significantly higher than those in standard-risk(SR) group(Pa