Behcet's Disease (BD) is a multisystem vasculitis characterized by disease alternated with recurrent episodes and remissions, involving genital, oral, ocular uvea, cutaneous, and articular manifestations. The nuclear factor (NF)-κB signaling pathway paly an important role in the BD progression. It encompasses diverse gene, protein, and cellular regulatory mechanisms operating across various levels, alongside microbiological and experimental studies involving animals and cells. At the protein research findings, activation of the NF-κB pathway in BD patients is marked by elevated plasma levels of soluble CD40 ligand, which stimulates neutrophils to release reactive oxygen species and extracellular traps, thereby promoting inflammation. At the cellular research findings, macrophages in BD patients polarize towards classically activated macrophages phenotype through the NF-κB pathway, exacerbating the inflammatory response. The activation of NF-κB is associated with increased expression of anti-apoptotic proteins in T cells, leading to prolonged inflammation. Microbiological investigations reveal that the decreased gut microbiota diversity in BD patients compromises intestinal barrier integrity. NF-κB pathway involvement in regulating neutrophil and type 1 helper T cell (Th) 1/Th17 cell function worsens inflammation. Genetically, BD patients exhibit polymorphisms in immune regulatory genes, which contribute to inflammation through the NF-κB pathway. Mutations in NF-κB-associated genes elevate the risk of BD, while mutations in the endogenous inhibitor A20 lead to abnormal NF-κB activity, sustaining inflammation. Animal experiments and in vitro experiments corroborate the efficacy of NF-κB inhibitors in attenuating inflammation. Targeting upstream inflammatory factors within the NF-κB pathway yields positive outcomes in BD patients. In summary, the NF-κB signaling pathway plays a pivotal role in the development of BD. Developing NF-κB inhibitors may open new avenues for treating BD. Further research is necessary to comprehensively elucidate the precise mechanisms by which NF-κB operates in the pathogenesis of BD, as well as its potential clinical applications in therapy.
Objective Series of compl icated molecule signal pathway are involved in the bone regeneration. To explore the possibil ity of nuclear factore kappa B (NF-κB) which is taken as the “key activation” during the fracture healing and provide the laboratory evidence for the gene therapy of nonunion or delayed union of fractures. Methods Thirtythree adult male Wistar rats (weighing 180-220 g) were selected and divided randomly into 4 groups: group A (the control group, n=3), the rigth lower segments of radius were injected with normal sal ine 0.3 mL for 7 days, once per day; group B (Bay 11-7082 injection group, n=6), the middle and distal radius were injected with normal sal ine containing 50 μmol/L NF- κB inhibitor Bay 11-7082 0.3 mL for 7 days, once per day; group C (fracture group, n=12), the right middle and distal radius were cut by a sharp scissors to form per fracture model; and group D (Bay 11-7082 treatment group, n=12), based on group C, 0.3 mL of 50 μmol/L Bay 11-7082 were injected into the fracture site for 7 days, once per day. The callus tissues were harvested at 3, 7, 14, and 28 days after fracture for Western blot analysis, alkal ine phosphatase (ALP) activity assessment, prostaglandins E2 (PGE2) production assay, and histological observation. Results The rats of all groups were survivaltill the experiment completion. At 3 and 7 days after injection, there was no significant difference in the ALP activity and PGE2 production between group B and group A (P gt; 0.05); but group C was significantly higher than group A (P lt; 0.01) and group D was significantly lower than group A (P lt; 0.01). The expressions of NF-κB p65, bone morphogenetic protein 7 (BMP-7), and inhibitor of DNA binding 2 (Id2) were observed at fracture sites of 4 groups. There was no significant difference in the expressions of NF-κB p65, BMP-7, and Id2 between group B and group A (P gt; 0.05); the expressions of NF-κB p65 and BMP-7 were significantly higher and the expression of Id2 was significantly lower in group C than group A (P lt; 0.01); and the expressions of NF-κB p65 and BMP-7 were significantly lower and the expression of Id2 was significantly higher in group D than group A (P lt; 0.01). The histological observation showed that a lot of osseous callus formed in group C at 14 and 28 days, but osseous callus just began to form in group D at 28 days. Conclusion NF-κB p65 can facil itate early fracture heal ing of rat radius by elevating the PGE2 production and regulating BMP-7 and Id2 expression.
Diabetic macular edema (DME) is one of the main reasons causing blindness in patients with diabetic retinopathy. In recent years, with the recognition of the pathogenic role of vascular endothelial growth factor (VEGF) in DME, many clinical trials of intravitreal injection of anti-VEGF drugs have been carried out at home and abroad, proving that it has significant effects in improving visual acuity and reducing macular edema, and has become the first-line treatment of DME. However, there are still many challenges in routine clinical application of anti-VEGF drugs, such as frequent injections, insensitivity to treatment, and it is unclear whether repeated injections will cause damage to retina. The pathophysiological process of DME is very complicated, in addition to VEGF, there are many inflammatory factors and growth factors involved. Clinical trials of long-acting anti-VEGF agents, drugs of other targets and gene therapy are also being carried out. It is believed that with the in-depth research and progress of clinical trials, the gradual application of anti-VEGF drugs, other drugs and therapy in clinical practice are just around the corner, which is expected to provide more convenient and effective treatments for DME patients in the future.
ObjectiveTo observe RNA-Seq analysis of gene expression profiling in human retinal vascular endothelial cells after anti-vascular endothecial growth factor (VEGF) treatment.MethodsCultured the retinal vascular endothelial cells in vitro and logarithmic growth phase cells were used for experiments. The cells were divided into VEGF group and VEGF combined with anti-VEGF drugs group. The VEGF group cells were treated with 50 ng/ml VEGF for 72 h to simulate the high VEGF survival conditions of vascular endothelial cells in diabetic retinopathy. VEGF combined with anti-VEGF drug group cells was treated with 50 ng/ml VEGF and 2.5 μg/ml anti-VEGF drugs for 72 h to imitate the microenvironment of cells following the anti-VEGF drugs treatment, and whole transcriptome sequencing approach was applied to the above two groups of cells through RNA-Seq. Now with biological big data obtained as a basis, to analyze the differentially expressed genes (DEGs). And through enrichment analysis to explain the differential functions of DEGs and their signal pathways.ResultsThe gene expression profiles of the two groups of cells were obtained. Through analysis, 328 DEGs were found, including 194 upregulated and 133 downregulated ones. The functions of DEGs were influenced by regulations over molecular biological process, cellular energy metabolism and protein synthesis, etc. Among these genes, SI,PRX and HPGD were related to protein synthesis, BIRCT to cellular apoptosis, and ABLIM1 and CRB2 to retinal development, and ABCG1, ABCA9 and ABCA12 were associated with the cholesterol of macrophage and the transfer of phospholipid. GO enrichment analysis showed that DEGs mainly act in three ways: regulating biological behavior, organizing cellular component and performing molecular function. Pathway enrichment analysis showed that gene expressions of the two cell groups were differentiated in ECM receptor pathway, and Notch, mitogen-activated protein kinase, transforming growth factor (TGF)-β and Wnt signal pathways. Among them, the gene expression in TGF-β signal pathway attracts most attention, where the DEGs, such as CAMK2B, COL3A1, CYGB, PTGER2 and HS6ST2, among others, were closely related to fibrosis process.ConclusionThe anti-VEGF drugs may enhance the expression of CAMK2B, COL3A1, CYGB, PTGER2 and others genes related to TGF-β signal pathway and aggravate retinal fibrosis disease.
Objective To evaluate the effect of recombinant human growth hormone(rhGH) on growth of human colonic cancer cells (COLO-320) in vitro. Methods Human COLO-320 cells in logarithm growing period were cultured for 24 h,48 h or 72 h with variant concentrations of rhGH,camptothecine (CPT) or rhGH combined CPT in calf serum(serum group) or calf serum-free (serum-free group). Light density of cells were determined by MTT method, so that cellular inhibition rate were calculated.Results No influence on cell growth or inhibition rate was observed from cultures with variant concentrations and different acting times of rhGH (P＞0.05). Inhibition rate of single CPT or CPT combined rhGH were much more increased than single rhGH used (P＜0.01) with no statistical significance (P＞0.05).Conclusion The results show that rhGH has neither direct COLO-320 cells stimulation nor any evidence of COLO-320 cells inhibition, and has no influence of CPT on COLO-320 cells inhibition in vitro.
Objective To study the effect of p38MAPK activity on tumor necrosis factor-α （TNF-α） mRNA and intercellular adhesion molecule 1 （ICAM1） mRNA expressions of isolated rabbit liver during early stage of cold preservation and reperfusion period. Methods Based on the cold preservation and reperfusion model of isolated rabbit liver, the animals were divided into inhibition group （n=12） with 3 μmol/L SB202190 (p38MAPK specificity inhibitor) in perfusate and control group （n=12） with no SB202190 in perfusate. Liver tissue samples were harvested at the time points of before resection, end of cold preservation, and different reperfusion period (10, 30, 60 and 120 min). Protein expression and activity of p38MAPK were detected by Western blot and immunoprecipitation respectively, expression of TNF-α mRNA was detected by RT-PCR, and expression of ICAM1 mRNA was detected by in situ hybridization. Results There was no obvious change of expression of p38MAPK protein in liver tissue both in two groups during the total period （P＞0.05）， and there was no statistically significant difference between two groups （P＞0.05）. At time points of end of cold preservation, 10, 30 and 60 min of reperfusion, the activity of p38MAPK in control group was significantly higher than that at the time points of before resection and 120 min of reperfusion (P＜0.01), and was also significantly higher than that in inhibition group at the same time points (P＜0.01). There was no significant difference in activity of p38MAPK among all time points in inhibition group (P＞0.05). The expressions of TNF-α mRNA and ICAM1 mRNA at the time points of before resection, end of cold preservation, and 10 and 30 min of reperfusion were significantly lower than those in 60 and 120 min of reperfusion in both two groups (P＜0.05, P＜0.01); The expressions of TNF-α mRNA and ICAM1 mRNA in inhibition group were significantly lower than those in control group at the time points of 60 and 120 min of reperfusion (P＜0.01). The activity of p38MAPK of liver tissue during cold preservation and reperfusion period was significantly correlated with the level of TNF-α mRNA and level of ICAM1 mRNA expression （r=0.996, P＜0.01； r=0.985, P＜0.01）. Conclusions These results suggest that p38MAPK pathway may regulate the expressions of TNF-α and ICAM1 at the level of transcription and the activation of p38MAPK can up-regulate TNF-α and ICAM1 expressions, which may be one of the important mechanisms to cause ischemia-reperfusion injury of isolated liver during cold preservation and reperfusion period.
Objective To review researches of the role of inhibitorof differentiation 2(Id2) in skeletal muscle regeneration. Methods The latest original literature concerning Id2 and its role in skeletal muscle regeneration was extensively reviewed. Results Id2 could form heterodimers by combining with E protein to prevent myogenic regulatory factors (MRFs) forming heterodimers by combining with E protein, to inhibit the transcription activity of MRFs anddifferentiation of skeletal muscle cell. Conclusion Id2 plays an important role in skeletal muscle regeneration.
ObjectiveTo detecte the expressions of phosphatase and tensin homolog deleted on chromosome ten (PTEN) gene and protein in peripheral blood mononuclear cells (PBMC) of patients with sepsis and to explore its role in the pathogenesis of acute obstructive suppurative cholangitis (AOSC). MethodsThe PBMC and serum were separated from AOSC patients (n=25) before treatment and in 1 week after recure, and healthy volunteers (normal control group, n=15). The serum levels of lipopolysaccharide (LPS), tumor necrosis factor-α (TNF-α) and interleukin 10 (IL-10) were detected by enzyme linked immunosorbent assay (ELISA) methods. The mRNA and protein expression levels of PTEN, nuclear fator κB P65 (NF-κB), and inhibitor of NF-κB (IκB) were detected by real-time quantitative polymerase chain reaction (qRT-PCR) and Western blot, respectively. ResultsThe levels of LPS, TNF-α, and IL-10 before treatment were significantly higher than those of normal control group (P < 0.05), the indicators were significantly decreased and close to normal levels in 1 week after recure. The mRNA and protein expression levels of PTEN and IκB before treatment were lower than those of normal control group (P < 0.05), NF-κB P65 was higher than those of normal control group (P < 0.05), while the phosphorylation levels of PTEN and IκB were higher than those normal control group (P < 0.05), and in 1 week after recure, the above indicators returned to normal levels. ConclusionsSepsis shift may be associated with the occurrence of intestinal LPS, and caused the imbalance between proinflammatory and anti-inflammatory cytokines in the body. PTEN for phosphorylation activation of IκB or directly activation of NF-κB participate the originating process of sepsis, hinting a therapeutic potentialities in the early stage of sepsis.
Objective To investigate the expression changes of nuclear factor kappa B (NF-κB) and matrix metalloproteinase-9 (MMP-9) in the cultured hepatocellular carcinoma cells 9204 (HCC9204) transfected with inhibitory kappa B alpha（IκB-α)vector. Methods After pcDNA3-IκB-α vector and pcDNA3 were transfected into HCC9204 by lipofectamine method, Western-blot and RT-PCR analysis were used to detect the expressions of NF-κB and MMP-9. Migration and invasion of tumor cells were assayed by fundus membrane invaded by them. Results When pcDNA3-IκB-α was transfected into HCC9204, the expression of NF-κB was decreased at the protein level, and the expression of MMP-9 mRNA and the invision and metastasis ability of transfected cells were obviously decreased. Conclusion When the activity of NF-κB is inhibited, the ability of invasion and metastasis in HCC9204 cells decrease, which could be related to the decreased the expression of MMP-9.
Objective To study the effect of genistein (Gen) in combination with 5-fluorouracil (5-FU) on human gastric carcinoma cells in vitro. Methods Human gastric carcinoma cell line SGC-7901 was t reated with Gen and 5-FU alone or in combination for 24 , 48 and 72 hours , respectively. MTT assay was used to investigate the inhibitory effect of Gen and 5-FU on SGC-7901 cells. Transmission electron microscopy was used to observe the ultramicrost ructure changes of cancer cells and the flow cytometry was used to detect the distribution of cell cycle. Results Gen and 5-FU alone or in combination inhibited the proliferation of SGC-7901 cells significantly in both time-dependent and dose-dependent manner. The combination of Gen with 5-FU had synergistic effect on the growth of SGC-7901 cell line when the inhibition ratio was 20 % to 80 %. When SGC-7901 cells exposed to 18. 8μmol/ L Gen , 8. 84μmol/ L 52FU , and 3. 06μmol/ L Gen combined with 7. 96μmol/ L 5-FU for 72 hours , 62. 97 % , 63. 76 % and 67. 46 % SGC-7901 cells were arrested in G2 / M , S and G0 / G1 phrase , respectively. Gen and 5-FU could both induce apoptosis and arrest cell cycle of SGC-7901 cells. Conclusion Combination therapy of Gen with 5-FU may take synergistic inhibitory effect on the proliferation of gastric cancer cells by resting cell cycle and inducing cell apoptosis at a certain range of concent ration.