Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 20 de 208
Filter
1.
Acta Pharmaceutica Sinica B ; (6): 340-354, 2021.
Article in English | WPRIM | ID: wpr-881140

ABSTRACT

Enormous studies have corroborated that long non-coding RNAs (lncRNAs) extensively participate in crucial physiological processes such as metabolism and immunity, and are closely related to the occurrence and development of tumors, cardiovascular diseases, nervous system disorders, nephropathy, and other diseases. The application of lncRNAs as biomarkers or intervention targets can provide new insights into the diagnosis and treatment of diseases. This paper has focused on the emerging research into lncRNAs as pharmacological targets and has reviewed the transition of lncRNAs from the role of disease coding to acting as drug candidates, including the current status and progress in preclinical research. Cutting-edge strategies for lncRNA modulation have been summarized, including the sources of lncRNA-related drugs, such as genetic technology and small-molecule compounds, and related delivery methods. The current progress of clinical trials of lncRNA-targeting drugs is also discussed. This information will form a latest updated reference for research and development of lncRNA-based drugs.

2.
J Biosci ; 2019 Jun; 44(2): 1-6
Article | IMSEAR | ID: sea-214371

ABSTRACT

Recent research has shown that cell-free chromatin (cfCh) particles that are released from the billions of cells that die in thebody everyday can enter into healthy cells, integrate into their genomes and induce dsDNA breaks and apoptotic responses.Genomic integration of cfCh activates NFjB suggesting a novel mechanism of induction of systemic inflammation. SinceDNA damage and inflammation are underlying pathologies in multiple devastating acute and chronic disease conditions,the discovery of agents that can inactivate cfCh may provide therapeutic possibilities.

3.
Article in Chinese | WPRIM | ID: wpr-755575

ABSTRACT

Objective To evaluate the effect of sevoflurane preconditioning on high-mobility group box 1 protein ( HMGB1) ∕Toll-like receptor 4 ( TLR4) ∕nuclear factor kappa B ( NF-κB) signaling pathway during lung ischemia-reperfusion ( I∕R) in rats. Methods Thirty-six clean-grade healthy male Sprague-Dawley rats, aged 8-10 weeks, weighing 200-250 g, were divided into 3 groups ( n=12 each) using a random number table method: sham operation group ( group S) , lung I∕R group ( group I∕R) and sevoflu-rane preconditioning group ( group SP ) . The right pulmonary hilum was only isolated but not ligated in group S. Lung I∕R was induced by clamping the right pulmonary hilum for 60 min followed by 120 min of reperfusion in anesthetized rats in group I∕R. In group SP, 2. 1% sevoflurane was inhaled for 30 min to per-form sevoflurane preconditioning, and the lung I∕R model was established at 10 min after the end of inhala-tion. The rats were sacrificed at 120 min of reperfusion, and the lungs were removed for examination of the pathological changes which were scored and for determination of wet to dry weight ratio ( W∕D ratio) , con-tent of tumor necrosis factor-alpha ( TNF-α) in lung tissues ( by enzyme-linked immunosorbent assay) and expression of HMGB1, TLR4 and NF-κB protein in lung tissues (by Western blot). Results Compared with group S, the pathological scores, W∕D ratio and content of TNF-α were significantly increased, and the expression of HMGB1, TLR4 and NF-κB was up-regulated in I∕R and SP groups ( P<0. 05) . Compared with group I∕R, the pathological scores, W∕D ratio and content of TNF-αwere significantly decreased, and the expression of HMGB1, TLR4 and NF-κB was down-regulated ( P<0. 05) , and the pathological changes of lung tissues were significantly attenuated in group SP . Conclusion Sevoflurane preconditioning reduces lung I∕R injury probably through inhibiting HMGB1∕TLR4∕NF-κB signaling pathway in rats.

4.
Article in Chinese | WPRIM | ID: wpr-709829

ABSTRACT

Objective To evaluate the effect of anti-myosin monoclonal antibodies-nuclear factor kappa B (NF-κB) decoy oligodeoxynucleotides-lipofectamine compound (mAb2G4-ODN-lip) on myocardial ischemia-reperfusion (I/R) injury in rats.Methods Forty clean-grade healthy adult male Sprague-Dawley rats,aged 8-10 weeks,weighing 240-260 g,were divided into 4 groups (n=10 each) using a random number table:sham operation group (group S),myocardial I/R group (group I/R),ODN-lip group (group ODN) and mAb2G4-ODN-lip group (group mAb2G4).Myocardial I/R was induced by occlusion of the left anterior descending branch of coronary artery for 30 min followed by 120 min reperfusion.In ODN and mAb2G4 groups,ODN-lip (100 μg ODN) and mAb2G4-ODN-lip (100 μg ODN) compounds were injected via the femoral vein,respectively,immediately after onset of ischemia.The left anterior descending branch of coronary artery was only occluded but not ligated in group S.The animals were sacrificed at 120 min of reperfusion and myocardial specimens of the left ventricle on the ischemic side were obtained for examination of the pathological changes (using haematoxylin and eosin staining) and for determination of the expression of NF-κB (by Western blot) and contents of tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6) (by enzyme-linked immunosorbent assay).Results Compared with group S,the expression of NF-κB was significantly up-regulated,and the contents of TNF-α and IL-6 were increased in I/R,ODN and mAb2G4 groups (P< 0.05).Compared with group I/R,the expression of NF-κB was significantly down-regulated,and the contents of TNF-α and IL-6 were decreased in ODN and mAb2G4 groups (P<0.05).Compared with group ODN,the expression of NF-κB was significantly down-regulated,and the contents of TNF-α and IL-6 were decreased in group mAb2G4 (P<0.05).The pathological changes of myocardial tissues were significantly attenuated in group I/R,group ODN and group mAb2G4 in turn.Conclusion mAb2G4-ODN-lip can mitigate myocardial I/R injury in rats.

5.
Chinese Journal of Geriatrics ; (12): 206-210, 2018.
Article in Chinese | WPRIM | ID: wpr-709221

ABSTRACT

Objective To investigate the mechanism that receptor activator of NF-κB ligand (RANKL) promotes arterial calcification.Methods Firstly,RANKL was added into the culture media,in which the monocyte precursor cells alone were cultured.Morphological observation and tartrate resistant acid phosphatase(TRAP)stain were used to assess whether RANKL could induce the monocyte precursor cells to differentiate into osteoclast-like cells.During arterial calcification,both in vivo and in vitro expressions of RANKL and osteoprotegerin (OPG,as RANKL inhibitor)were measured via real-time PCR.The extent of osteoclast-like cell differentiation was also assessed.Results It was found that RANKL could induce osteoclast-like cell differentiation.There were no both in vivo and in vitro expressions of osteoclast-like cells in the early stage of calcification.At that time,the ratio of RANKL to OPG was very low.In the late stage of calcification,a small amount of osteoclast-like cell expression coincided with a relatively high ratio of RANKL to OPG.According to the results,the ratio of RANKL to OPG was very low during most of the arterial calcification period.This made it possible for OPG to completely inhibit RANKL-induced osteoclast-like cell differentiation.The ratio of RANKL to OPG was (0.36 ± 0.08) (F =36) and (1.68 ± 0.08) (F =36) respectively in the early and late subgroup of calcification group in the animal model,but was zero in the control group(both P<0.05).The ratio of RANKL to OPG was(0.42±0.09) (F=16)and(1.50 ± 0.10)(F=16)respectively in the early and late subgroup of calcification group in the cell model,but was zero in the control group(both P<0.05).Conclusions Our result likely explains why RANKL has the ability to induce osteoclast-like cell differentiation,but acts as a promoter of calcification.

6.
Chinese Journal of Endemiology ; (12): 181-185, 2018.
Article in Chinese | WPRIM | ID: wpr-701295

ABSTRACT

Objective To clarify the role of nuclear factor κB(NF-κB) signaling pathway in pathogenesis of Kashin-Beck disease(KBD) by observing the expression of NF-κB p65 in the whole blood samples of patients with KBD and controls,and the expression of NF-κB p65 in C28/I2 chondrocyte, and to analyze the role of NF-κB p65 molecule in chondrocyte apoptosis. Methods Through a case-control study, 161 patients with KBD (KBD group) were selected from Xunyi, Yongshou, Changwu, Linyou, Qianyang and Long counties in KBD endemic areas and 312 healthy people(control group) were matched by age and sex in Shaanxi Province. Venous blood samples were collected from patients and healthy controls, which were anticoagulated and used for determination of NF-κB p65 protein.According to the group design,the model of C28/I2 chondrocyte oxidative damage was established.The experiments were divided into 4 groups including control group(C), tBHP injury group (O, tBHP 300.00 μmol/L), low selenium pre-protection group (OS1, 0.05 mg/L Na2SeO3+ 300.00 μmol/L tBHP), and middle selenium pre-protection group(OS2, 0.10 mg/L Na2SeO3+ 300.00 μmol/L tBHP). Then, cell apoptosis was detected by Hoechst 33342 and reactive oxygen species (ROS) was detected by dichlorfluorescein(DCF) method. The protein was extracted by Trizol method, then protein expression level of NF-κB p65 molecule was detected by Western blotting in whole blood samples and C28/I2 chondrocyte. Results The differences in age and sex were not statistically significant between KBD group and control group (t = 0.336, P > 0.05; χ2= 0.407, P > 0.05). The protein expression level of NF-κB p65 in KBD group was 1.835 times as high as that of control group (KBD:0.167 ± 0.026, control: 0.091 ± 0.014, t = 5.147, P < 0.01). Under the fluorescence microscope, chondrocyte showed strong blue fluorescence in tBHP group and the level of ROS(1.219 ± 0.104) was higher than those of low and middle selenium pre-protection groups(0.832 ± 0.077, 0.635 ± 0.070, P < 0.05).The protein expression level of NF-κB p65 in tBHP group (1.563 ± 0.351) was higher than that of control group (0.451 ± 0.069, P < 0.05), and protein levels of NF-κB p65 had a decreasing tendency in low and middle selenium pre-protection groups compared to tBHP group. Conclusion The NF-κB signaling pathway is up-regulated in KBD patients, moreover, chondrocyte experiments show that cell apoptosis is mediated via upregulation of NF-κB p65,which suggests NF-κB signaling pathway may play an important role in pathogenesis of KBD.

7.
Chinese Journal of Digestion ; (12): 250-257, 2018.
Article in Chinese | WPRIM | ID: wpr-711593

ABSTRACT

Objective To investigate the effects of caspase recruitment domain-containing protein 9 (CARD9)expression in peritoneal macrophages on severe acute pancreatitis(SAP)in rats and its mechanism.Methods A total of 60 male Sprague Dawley rats were divided into control group(n=6), SAP group(n=18),small interfering RNA(siRNA)control group(n=18)and siRNA CARD9 group (n=18).SAP rat models were established.At three,six and twelve hours after the models were established,ascites was collected,peritoneum was lavaged and peritoneal macrophages were isolated and cultured.The expressions of CARD9,nuclear factor-kappaB(NF-κB),p38 mitogen-activated protein kinase(p38MA PK)at mRNA level in peritoneal macrophages was measured by real-time polymerase chain reaction(RT-PCR).The levels of tumor necrosis factor-α(TNF-α),interleukin(IL)-1β and IL-6 in peripheral blood were detected by enzyme-linked immunosorbent assay(ELISA).LSD or Tamhane′s T2 methods were performed for statistical analysis.Results At three,six and twelve hours after the models were established,CA RD9 mRNA levels of peritoneal macrophages in SAP group were 1.63 ± 0.05,1.68 ± 0.24 and 2.61 ± 0.02,respectively,which were all higher than that of control group(1.01 ± 0.23),and the differences were statistically significant(t=25.97,6.86 and 131.59;all P<0.05);the levels of CA RD9 mRNA of siRNA CARD9 group were 1.45 ± 0.02,1.24 ± 0.03 and 1.63 ± 0.03,respectively,which were lower than that of SAP group at the same time points,and the differences were statistically significant(t=-7.81,-4.46 and -62.13;all P< 0.05).At three,six and twelve hours after the models were established,the mRNA levels of NF-κB and p38MA PK of peritoneal macrophages of rats in SAP group were 1.51 ± 0.08,1.81 ± 0.10,2.30 ± 0.05 and 1.37 ± 0.13,1.69 ± 0.18,2.42 ± 0.23,respectively, which were higher than those of control group(1.00 ± 0.01,1.03 ± 0.08),and the differences were statistically significant(tNF-κB=15.10,19.95 and 60.36;tp38MAPK=5.37,8.34 and 14.11;all P<0.05);the levels of N F-κB mRNA in siRNA CARD9 group were 1.38 ± 0.05,1.57 ± 0.06 and 1.76 ± 0.09, respectively,which were lower than that of SAP group at the same time points,and the differences were statistically significant(t= -3.32,-5.07 and -12.70;all P<0.05).At six and twelve hours after the models were established,the p38MA PK mRNA levels of siRNA CARD9 group were 1.50 ± 0.10 and 2.00 ± 0.09,respectively,which were lower than that of SAP group,and the differences were statistically significantly(t= -2.30 and -4.17,both P< 0.05).At three,six and twelve hours after the models were established,the levels of TNF-α,IL-1β and IL-6 in peripheral blood of SAP group were(53.49 ± 21.64)pg/mL,(108.62 ± 22.76)pg/mL and(139.00 ± 15.35)pg/mL;(43.86 ± 18.30)pg/mL, (87.51 ± 17.10)pg/mL and(117.27 ± 14.57)pg/mL;(78.38 ± 32.70)pg/mL,(156.39 ± 30.56)pg/mL and(209.56 ± 26.09)pg/mL,respectively,which were higher than those of control group((2.79 ± 1.17),(7.13 ± 4.52),(12.73 ± 8.08)pg/mL),and the differences were statistically significant(tTNF-α=5.73,11.37 and 21.69;tIL-1β=4.77,11.13 and 17.68;tIL-6=4.77,11.32 and 17.68;all P<0.05).At six and twelve hours after the models were established,the levels of TNF-α,IL-1β and IL-6 of siRNA CARD9 group were(75.73 ± 16.93)pg/mL,(108.23 ± 14.02)pg/mL;(63.05 ± 11.98)pg/mL, (91.56 ± 14.28)pg/mL and(112.67 ± 21.40)pg/mL,(163.62 ± 25.51)pg/mL,respectively,which were lower than those of SAP group,and the differences were statistically significant(tTNF-α= -2.84,-3.63;tIL-1β= -2.88,-3.09;tIL-6= -2.88,-3.09;all P< 0.05).Conclusions There are CARD9-related NF-κB and p38MAPK pathways in peritoneal macrophages of SAP rats.Intervention of the expression of CARD9 in peritoneal macrophages,especially at early stage of SAP may relieve the inflammation reaction and pancreatic injury,which may provide a new method for SAP treatment.

8.
Practical Oncology Journal ; (6): 43-47, 2017.
Article in Chinese | WPRIM | ID: wpr-507141

ABSTRACT

In eukaryotic cells ,NF-κB transcription factor family regulates many processes like cell sur-vival,growth,and apoptosis.It participates in the development of a variety of diseases ,including inflammatory,im-mune disease ,and cancer .As an inflammatory factor , NF-κB mediates the transformation of chronic colitis to cancer during the course of colorectal cancer .Furthermore,it could inhibit cell apoptosis through regulating cell cycles,which promotes the development of colorectal cancer and mediates the multidrug resistance of the tumor cells.Therefore,targeting NF-κB,a large number of preparations involved both Chinese and western medicine has been researched .The further research and the use of them may be an effective method to cure the colorectal cancer in clinical work .

9.
Article in English | WPRIM | ID: wpr-57551

ABSTRACT

BACKGROUND: Lipocalin-2 (LCN2), a small glycoprotein, has a pivotal role in diverse biological processes such as cellular proliferation and differentiation. We previously reported that LCN2 is implicated in osteoclast formation induced by receptor activator of nuclear factor-kappa B ligand (RANKL) and macrophage colony-stimulating factor (M-CSF). In the present study, we used a knockout mouse model to further investigate the role of LCN2 in osteoclast development. METHODS: Osteoclastogenesis was assessed using primary bone marrow-derived macrophages. RANKL and M-CSF signaling was determined by immunoblotting, cell proliferation by bromodeoxyuridine (BrdU) enzyme-linked immunosorbent assay (ELISA), and apoptosis by cell death detection ELISA. Bone morphometric parameters were determined using a micro-computed tomography system. RESULTS: Our results showed that LCN2 deficiency increases tartrate-resistant acid phosphatase (TRAP)-positive multinucleated osteoclast formation in vitro, a finding that reflects enhanced proliferation and differentiation of osteoclast lineage cells. LCN2 deficiency promotes M-CSF-induced proliferation of bone marrow macrophages (BMMs), osteoclast precursors, without altering their survival. The accelerated proliferation of LCN2-deficient precursors is associated with enhanced expression and activation of the M-CSF receptor, c-Fms. Furthermore, LCN2 deficiency stimulates the induction of c-Fos and nuclear factor of activated T cells c1 (NFATc1), key transcription factors for osteoclastogenesis, and promotes RANKL-induced inhibitor of kappa B (IkappaBalpha) phosphorylation. Interestingly, LCN2 deficiency does not affect basal osteoclast formation in vivo, suggesting that LCN2 might play a role in the enhanced osteoclast development that occurs under some pathological conditions. CONCLUSIONS: Our study establishes LCN2 as a negative modulator of osteoclast formation, results that are in accordance with our previous findings.


Subject(s)
Acid Phosphatase , Animals , Apoptosis , Biological Phenomena , Bone Marrow , Bromodeoxyuridine , Cell Death , Cell Proliferation , Enzyme-Linked Immunosorbent Assay , Glycoproteins , Immunoblotting , Macrophage Colony-Stimulating Factor , Macrophages , Mice , Mice, Knockout , NF-kappa B , Osteoclasts , Phosphorylation , RANK Ligand , T-Lymphocytes , Transcription Factors
10.
Article in English | WPRIM | ID: wpr-20742

ABSTRACT

Lichens have been known to possess multiple biological activities, including anti-proliferative and anti-inflammatory activities. Vascular cell adhesion molecule-1 (VCAM-1) may play a role in the development of atherosclerosis. Hence, VCAM-1 is a possible therapeutic target in the treatment of the inflammatory disease. However, the effect of lobaric acid on VCAM-1 has not yet been investigated and characterized. For this study, we examined the effect of lobaric acid on the inhibition of VCAM-1 in tumor necrosis factor-alpha (TNF-alpha)-stimulated mouse vascular smooth muscle cells. Western blot and ELISA showed that the increased expression of VCAM-1 by TNF-alpha was significantly suppressed by the pre-treatment of lobaric acid (0.1-10 mug/ml) for 2 h. Lobaric acid abrogated TNF-alpha-induced NF-kappaB activity through preventing the degradation of IkappaB and phosphorylation of extracellular signal-regulated kinases (ERK), c-Jun N-terminal kinases (JNK), and p38 mitogen activated protein (MAP) kinase. Lobaric acid also inhibited the expression of TNF-alpha receptor 1 (TNF-R1). Overall, our results suggest that lobaric acid inhibited VCAM-1 expression through the inhibition of p38, ERK, JNK and NF-kappaB signaling pathways, and downregulation of TNF-R1 expression. Therefore, it is implicated that lobaric acid may suppress inflammation by altering the physiology of the atherosclerotic lesion.


Subject(s)
Animals , Atherosclerosis , Blotting, Western , Down-Regulation , Enzyme-Linked Immunosorbent Assay , Extracellular Signal-Regulated MAP Kinases , Inflammation , Lichens , Mice , Muscle, Smooth, Vascular , NF-kappa B , Phosphorylation , Phosphotransferases , Physiology , Tumor Necrosis Factor-alpha , Vascular Cell Adhesion Molecule-1
11.
Article in English | WPRIM | ID: wpr-728547

ABSTRACT

(E)-3-(3-methoxyphenyl)-1-(2-pyrrolyl)-2-propenone (MPP) is an aldol condensation product resulting from pyrrole-2-carbaldehyde and m- and p- substituted acetophenones. However, its biological activity has not yet been evaluated. Since it has been reported that some propenone-type compounds display anti-inflammatory activity, we investigated whether MPP could negatively modulate inflammatory responses. To do this, we employed lipopolysaccharide (LPS)-stimulated macrophage-like RAW264.7 cells and examined the inhibitory levels of nitric oxide (NO) production and transcriptional activation, as well as the target proteins involved in the inflammatory signaling cascade. Interestingly, MPP was found to reduce the production of NO in LPS-treated RAW264.7 cells, without causing cytotoxicity. Moreover, this compound suppressed the mRNA levels of inflammatory genes, such as inducible NO synthase (iNOS) and tumor necrosis factor (TNF)-alpha. Using luciferase reporter gene assays performed in HEK293 cells and immunoblotting analysis with nuclear protein fractions, we determined that MPP reduced the transcriptional activation of nuclear factor (NF)-kappaB. Furthermore, the activation of a series of upstream signals for NF-kappaB activation, composed of Src, Syk, Akt, and IkappaBalpha, were also blocked by this compound. It was confirmed that MPP was able to suppress autophosphorylation of overexpressed Src and Syk in HEK293 cells. Therefore, these results suggest that MPP can function as an anti-inflammatory drug with NF-kappaB inhibitory properties via the suppression of Src and Syk.


Subject(s)
Acetophenones , Genes, Reporter , HEK293 Cells , Immunoblotting , Luciferases , Macrophages , NF-kappa B , Nitric Oxide , Nitric Oxide Synthase , Nuclear Proteins , RNA, Messenger , Transcriptional Activation , Tumor Necrosis Factor-alpha
12.
Article in English | WPRIM | ID: wpr-727995

ABSTRACT

Adult hippocampal dentate granule neurons are generated from neural stem cells (NSCs) in the mammalian brain, and the fate specification of adult NSCs is precisely controlled by the local niches and environment, such as the subventricular zone (SVZ), dentate gyrus (DG), and Toll-like receptors (TLRs). Epigallocatechin-3-gallate (EGCG) is the main polyphenolic flavonoid in green tea that has neuroprotective activities, but there is no clear understanding of the role of EGCG in adult neurogenesis in the DG after neuroinflammation. Here, we investigate the effect and the mechanism of EGCG on adult neurogenesis impaired by lipopolysaccharides (LPS). LPS-induced neuroinflammation inhibited adult neurogenesis by suppressing the proliferation and differentiation of neural stem cells in the DG, which was indicated by the decreased number of Bromodeoxyuridine (BrdU)-, Doublecortin (DCX)- and Neuronal Nuclei (NeuN)-positive cells. In addition, microglia were recruited with activatingTLR4-NF-kappaB signaling in the adult hippocampus by LPS injection. Treating LPS-injured mice with EGCG restored the proliferation and differentiation of NSCs in the DG, which were decreased by LPS, and EGCG treatment also ameliorated the apoptosis of NSCs. Moreover, pro-inflammatory cytokine production induced by LPS was attenuated by EGCG treatment through modulating the TLR4-NF-kappaB pathway. These results illustrate that EGCG has a beneficial effect on impaired adult neurogenesis caused by LPSinduced neuroinflammation, and it may be applicable as a therapeutic agent against neurodegenerative disorders caused by inflammation.


Subject(s)
Adult , Animals , Apoptosis , Brain , Bromodeoxyuridine , Dentate Gyrus , Hippocampus , Humans , Inflammation , Lipopolysaccharides , Mice , Microglia , Neural Stem Cells , Neurodegenerative Diseases , Neurogenesis , Neurons , Tea , Toll-Like Receptors
13.
Yonsei Medical Journal ; : 260-264, 2016.
Article in English | WPRIM | ID: wpr-220773

ABSTRACT

The epithelial cytokine response, associated with reactive oxygen species (ROS), is important in Helicobacter pylori (H. pylori)-induced inflammation. H. pylori induces the production of ROS, which may be involved in the activation of mitogen-activated protein kinases (MAPK), janus kinase/signal transducers and activators of transcription (Jak/Stat), and oxidant-sensitive transcription factor, nuclear factor kappa-light-chain-enhancer of activated B cells (NF-kappaB), and thus, expression of interleukin-8 (IL-8) in gastric epithelial cells. alpha-lipoic acid, a naturally occurring thiol compound, is a potential antioxidant. It shows beneficial effects in treatment of oxidant-associated diseases including diabetes. The present study is purposed to investigate whether alpha-lipoic acid inhibits expression of inflammatory cytokine IL-8 by suppressing activation of MAPK, Jak/Stat, and NF-kappaB in H. pylori-infected gastric epithelial cells. Gastric epithelial AGS cells were pretreated with or without alpha-lipoic acid for 2 h and infected with H. pylori in a Korean isolate (HP99) at a ratio of 300:1. IL-8 mRNA expression was analyzed by RT-PCR analysis. IL-8 levels in the medium were determined by enzyme-linked immunosorbent assay. NF-kappaB-DNA binding activity was determined by electrophoretic mobility shift assay. Phospho-specific and total forms of MAPK and Jak/Stat were assessed by Western blot analysis. ROS levels were determined using dichlorofluorescein fluorescence. As a result, H. pylori induced increases in ROS levels, mRNA, and protein levels of IL-8, as well as the activation of MAPK [extracellular signal-regulated kinase 1/2 (ERK1/2), c-Jun NH2-terminal kinase 1/2 (JNK1/2), p38], Jak/Stat (Jak1/2, Stat3), and NF-kappaB in AGS cells, which was inhibited by alpha-lipoic acid. In conclusion, alpha-lipoic acid may be beneficial for prevention and/or treatment of H. pylori infection-associated gastric inflammation.


Subject(s)
Enzyme-Linked Immunosorbent Assay , Epithelial Cells/metabolism , Gastric Mucosa/drug effects , Gene Expression Regulation, Bacterial , Helicobacter Infections/immunology , Helicobacter pylori/drug effects , Humans , Interleukin-8/genetics , JNK Mitogen-Activated Protein Kinases , Janus Kinase 1 , Mitogen-Activated Protein Kinases/biosynthesis , NF-kappa B/metabolism , RNA, Messenger/isolation & purification , Reactive Oxygen Species/metabolism , STAT3 Transcription Factor , Stomach/metabolism , Thioctic Acid/pharmacology
14.
Chinese Journal of Geriatrics ; (12): 898-900, 2015.
Article in Chinese | WPRIM | ID: wpr-482905

ABSTRACT

Objective To investigate the mechanism of homocysteine-induced microglia (BV-2 cells) expression of interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α).Methods The BV-2 cells were divided into blank control group,cysteine (Cys) group,homocysteine (Hcy) group and homocysteine and glutathione (Hcy+GSH) group,and the BV-2 cells in these groups were incubated with cysteine or homocysteine or homocysteine and glutathione together for 72 h.The mRNA expressions of IL-1β and TNF-α were assessed by RT-qPCR.The protein expressions of IL-1β and TNF-α in supernatant were detected by enzyme linked immunosorbent assay (ELISA).Western blot was used to observe the changes of NF-κB/p65 expression.Results There were significant differences in mRNA and protein expressions of IL-1β and TNF-α and NF-κB/p65 protein expression between groups (F=48.63,130.76,702.91,293.69,212.06,respectively,all P=0.000).The secretions of IL-1β and TNF-α were significantly improved by homocysteine (P<0.05),and were reversed by the treatment with glutathione (P<0.05).Western blot assay result showed that NF-κB/p65 was elevated after treatment with homocysteine (P<0.05).Conclusions Homocysteine can induce microglia expression of interleukin-1β and tumor necrosis factor-α,and NF-κB signaling pathway may be involved in this process.

15.
Chinese Journal of Digestion ; (12): 624-629, 2015.
Article in Chinese | WPRIM | ID: wpr-479263

ABSTRACT

Objective To investigate the mechanism of caspase recruitment domain‐containing protein 9 (CARD9) in the early stage of acute pancreatitis(AP) .Methods Peripheral blood mononuclear cells (PBMC ) of 49 AP patients (33 mild acute pancreatitis (MAP ) patients and 16 severe acute pancreatitis (SAP) patients) were collected on the Day 1st ,3rd and 5th of hospitalization .Twenty healthy volunteers were enrolled in control group .The expression level of CARD9 ,B‐cell lymphoma(Bcl)‐10 ,p38 mitogen‐activated protein kinase (MAPK ) and p65 nuclear factor Kappa B (NF‐κB ) in PBMC of AP patients were detected by Western blotting .The co‐localization ,expression and binding between CARD9 and Bcl‐10 in PBMC of control group ,SAP group and MAP group on the Day 1st hospitalization were determined by cell immune‐fluorescence staining and co‐immuno precipitation method .Single factor analysis of variance and Mann‐Whitney test were performed for data comparison between groups .Pearson method was used for correlation analysis .Results The results of Western blotting indicated that the expression of CARD9 and Bcl‐10 in PBMC of SAP group on the Day 1st ,3rd and 5th of hospitalization (1 .12 ± 0 .05 ,1 .03 ± 0 .03 and 1 .01 ± 0 .01 ;1 .74 ± 0 .08 ,1 .72 ± 0 .10 and 1 .69 ± 0 .11) were all significantly higher than those of control group (0 .33 ± 0 .10 and 1 .02 ± 0 .11) and MAP group (0 .71 ± 0 .02 ,0 .55 ± 0 .06 and 0 .25 ± 0 .07 ;1 .15 ± 0 .03 ,1 .09 ± 0 .07 and 1 .01 ± 0 .04) ,and the differences were statistically significant (F= 35 .76 and 18 .20 ,all P< 0 .05) .The expression of p38 MAPK in PBMC of SAP group on the Day lst ,3rd of hospitalization (1 .88 ± 0 .08 、1 .68 ± 0 .11) were significantly higher than those of MAP group on the Day 1st ,3rd ,5th (0 .86 ± 0 .08 ,0 .77 ± 0 .10 ,0 .73 ± 0 .20) and healthy control group (0 .58 ± 0 .24 , F= 7 .24 ,all P < 0 .01) .The expression of p65 NF‐κB in PBMC of SAP group on the Day 1st ,3rd of hospitalization (1 .64 ± 0 .02 ,1 .55 ± 0 .03) were significantly higher than those of MAP group on the Day 3rd ,5th (1 .06 ± 0 .14 ,0 .87 ± 0 .20) and healthy control group (1 .17 ± 0 .13 ,F= 4 .51 ,all P< 0 .05) .The results of immune‐fluorescence staining indicated that CARD9 and Bcl‐10 co‐localized in nucleus .The results of co‐immuno precipitation showed that the binding degree between CARD9 and Bcl‐10 of SAP group was significantly higher than that of control group and MAP group . Pearson correlative analysis suggested that the level of p65 NF‐κB and p38 MAPK in PBMC of AP patients were positive correlated with the expression of CARD9 (r= 0 .692 and 0 .834 ,both P< 0 .01) .Conclusion CARD9 is positive correlated with NF‐κB and MAPK , which indicates CARD9 induced inflammatory cytokines by activating NF‐κB and MAPK signaling pathways in AP .

16.
Article in Chinese | WPRIM | ID: wpr-460612

ABSTRACT

Diffuse large B-cell lymphoma (DLBCL) is the most common malignant lymphoma in China,which mostly starting from the lymph nodes.The studies find that the expression of NLRC5,member of nucleotide-binding oligomerization domain-like receptor (NLR) family,is evident in the lymphoid cells,which is likely to be an important basis for lymphocyte tumorigenesis.NLRC5 can block the nuclear factor-κB (NF-κB),a core component of signal transduction pathway,and affect the development of tumor.While continuing activation of NF-κB is a necessary condition for DLBCL cell survival.Therefore,NLRC5 is likely to inhibit excessive inflammation,thereby inhibiting the function of DLBCL,which promises to be a new target for immunotherapy treatment of DLBCL.

17.
Article in English | WPRIM | ID: wpr-96084

ABSTRACT

BACKGROUND: Arginine vasopressin (AVP) is widely used as a vasopressor agent. Some recent studies have suggested that AVP may exert an immunomodulatory effect. However, the mechanism about the anti-inflammatory effect of AVP is not well known. We investigated the effect of AVP on the ihibitor of kappa B (IkappaBalpha)/nuclear factor-kappa B (NF-kappaB) pathway in RAW 264.7 cells. METHODS: Cultured RAW 264.7 cells were pretreated with AVP and stimulated with lipopolysaccharide (LPS). To evaluate the effect of AVP on inflammatory cytokines, the concentration of interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha) were assessed by an enzyme-linked immunosorbent assay technique. The expression of IkappaBalpha and nuclear translocation of NF-kappaB p65 were measured by Western blotting, and IkappaB kinase (IKK) activity was analyzed by an in vitro immune complex kinase assay. To confirm the AVP effect on IkappaBalpha/NF-kappaB cascade and via V2 receptor, we added tolvaptan (V2 receptor antagonist) after AVP pretreatment. RESULTS: The increase of IL-6 and TNF-alpha in LPS-stimulated RAW 264.7 cells was suppressed by a treatment with AVP. Pretreatment of AVP inhibited increasing of IKK activity and IkappaBalpha degradation induced by LPS in RAW 264.7 cells. Furthermore, LPS induced and NF-kappaB transcription was inhibited by AVP pretreatment. The observed changes in IKK activity, IkappaBalpha degradation and NF-kappaB transcription by AVP was abolished by tolvaptan treatment. CONCLUSIONS: Our results suggest that AVP showed anti-inflammatory effect on LPS-induced IkappaBalpha/NF-kappaB cascade in mouse macrophages via V2 receptors.


Subject(s)
Animals , Antigen-Antibody Complex , Arginine Vasopressin , Blotting, Western , Cytokines , Enzyme-Linked Immunosorbent Assay , I-kappa B Kinase , Interleukin-6 , Macrophages , Mice , NF-kappa B , Phosphotransferases , Receptors, Vasopressin , Tumor Necrosis Factor-alpha
18.
Intestinal Research ; : 233-241, 2015.
Article in English | WPRIM | ID: wpr-96060

ABSTRACT

BACKGROUND/AIMS: Balsalazide is an anti-inflammatory drug used in the treatment of inflammatory bowel disease. Balsalazide can reduce inflammatory responses via several mechanisms, including inhibition of nuclear factor-kappaB (NF-kappaB) activity. Parthenolide (PT) inhibits NF-kappaB and exerts promising anticancer effects by promoting apoptosis. The present investigated the antitumor effects of balsalazide, combined with PT, on NF-kappaB in a representative human colorectal carcinoma cell line, HCT116. METHODS: We counted cells and conducted annexin-V assays and cell cycle analysis to measure apoptotic cell death. Western blotting was used investigate the levels of proteins involved in apoptosis. RESULTS: PT and balsalazide produced synergistic anti-proliferative effects and induced apoptotic cell death. The combination of balsalazide and PT markedly suppressed nuclear translocation of the NF-kappaB p65 subunit and the phosphorylation of inhibitor of NF-kappaB. Moreover, PT and balsalazide dramatically enhanced NF-kappaB p65 phosphorylation. Apoptosis, through the mitochondrial pathway, was confirmed by detecting effects on Bcl-2 family members, cytochrome c release, and activation of caspase-3 and -8. CONCLUSIONS: Combination treatment with PT and balsalazide may offer an effective strategy for the induction of apoptosis in HCT116 cells.


Subject(s)
Apoptosis , Blotting, Western , Caspase 3 , Cell Cycle , Cell Death , Cell Line , Colorectal Neoplasms , Cytochromes c , HCT116 Cells , Humans , Inflammatory Bowel Diseases , NF-kappa B , Phosphorylation
19.
Yonsei Medical Journal ; : 862-866, 2015.
Article in English | WPRIM | ID: wpr-137565

ABSTRACT

Helicobacter pylori (H. pylori) induces the activation of nuclear factor-kB (NF-kappaB) and cytokine expression in gastric epithelial cells. The Janus kinase/signal transducers and activators of transcription (Jak/Stat) cascade is the inflammatory signaling in various cells. The purpose of the present study is to determine whether H. pylori-induced activation of NF-kappaB and the expression of interleukin-8 (IL-8) are mediated by the activation of Jak1/Stat3 in gastric epithelial (AGS) cells. Thus, gastric epithelial AGS cells were infected with H. pylori in Korean isolates (HP99) at bacterium/cell ratio of 300:1, and the level of IL-8 in the medium was determined by enzyme-linked immonosorbent assay. Phospho-specific and total forms of Jak1/Stat3 and IkappaBalpha were assessed by Western blot analysis, and NF-kappaB activation was determined by electrophoretic mobility shift assay. The results showed that H. pylori induced the activation of Jak1/Stat3 and IL-8 production, which was inhibited by a Jak/Stat3 specific inhibitor AG490 in AGS cells in a dose-dependent manner. H. pylori-induced activation of NF-kappaB, determined by phosphorylation of IkappaBalpha and NF-kappaB-DNA binding activity, were inhibited by AG490. In conclusion, Jak1/Stat3 activation may mediate the activation of NF-kappaB and the expression of IL-8 in H. pylori-infected AGS cells. Inhibition of Jak1/Stat3 may be beneficial for the treatment of H. pylori-induced gastric inflammation, since the activation of NF-kappaB is inhibited and inflammatory cytokine expression is suppressed.


Subject(s)
Blotting, Western , DNA, Bacterial/analysis , Epithelial Cells/metabolism , Gastric Mucosa/drug effects , Gene Expression Regulation/drug effects , Gene Expression Regulation, Bacterial , Helicobacter Infections/immunology , Helicobacter pylori/genetics , Humans , Interleukin-8/genetics , Janus Kinase 1 , NF-kappa B/biosynthesis , Phosphorylation , RNA, Messenger/metabolism , STAT3 Transcription Factor , Signal Transduction/genetics
20.
Yonsei Medical Journal ; : 862-866, 2015.
Article in English | WPRIM | ID: wpr-137564

ABSTRACT

Helicobacter pylori (H. pylori) induces the activation of nuclear factor-kB (NF-kappaB) and cytokine expression in gastric epithelial cells. The Janus kinase/signal transducers and activators of transcription (Jak/Stat) cascade is the inflammatory signaling in various cells. The purpose of the present study is to determine whether H. pylori-induced activation of NF-kappaB and the expression of interleukin-8 (IL-8) are mediated by the activation of Jak1/Stat3 in gastric epithelial (AGS) cells. Thus, gastric epithelial AGS cells were infected with H. pylori in Korean isolates (HP99) at bacterium/cell ratio of 300:1, and the level of IL-8 in the medium was determined by enzyme-linked immonosorbent assay. Phospho-specific and total forms of Jak1/Stat3 and IkappaBalpha were assessed by Western blot analysis, and NF-kappaB activation was determined by electrophoretic mobility shift assay. The results showed that H. pylori induced the activation of Jak1/Stat3 and IL-8 production, which was inhibited by a Jak/Stat3 specific inhibitor AG490 in AGS cells in a dose-dependent manner. H. pylori-induced activation of NF-kappaB, determined by phosphorylation of IkappaBalpha and NF-kappaB-DNA binding activity, were inhibited by AG490. In conclusion, Jak1/Stat3 activation may mediate the activation of NF-kappaB and the expression of IL-8 in H. pylori-infected AGS cells. Inhibition of Jak1/Stat3 may be beneficial for the treatment of H. pylori-induced gastric inflammation, since the activation of NF-kappaB is inhibited and inflammatory cytokine expression is suppressed.


Subject(s)
Blotting, Western , DNA, Bacterial/analysis , Epithelial Cells/metabolism , Gastric Mucosa/drug effects , Gene Expression Regulation/drug effects , Gene Expression Regulation, Bacterial , Helicobacter Infections/immunology , Helicobacter pylori/genetics , Humans , Interleukin-8/genetics , Janus Kinase 1 , NF-kappa B/biosynthesis , Phosphorylation , RNA, Messenger/metabolism , STAT3 Transcription Factor , Signal Transduction/genetics
SELECTION OF CITATIONS
SEARCH DETAIL