Is Trimethylamine N-oxide (TMAO) Associated with NF-κB mRNA Expression in Patients with Coronary Artery Disease?

Abstract Background: Trimethylamine N-oxide (TMAO), a gut microbiota metabolite, is associated with cardiovascular disease (CVD) development. TMAO can trigger an inflammatory response by inducing the nuclear factor-kappa B (NF-κB) signaling cascade and increasing the expression of pro-inflammatory cytokines, contributing to the worsening of CVD. This study aimed to evaluate the association between TMAO plasma levels and inflammation in patients with coronary artery disease (CAD). Methods: A cross-sectional study was carried out including 29 patients with CAD. Peripheral blood mononuclear cells (PBMC) were isolated from fasting blood samples, and NF-κB and vascular cell adhesion protein 1 (VCAM1) mRNA expression were estimated using real-time quantitative PCR. We determined TMAO plasma levels by LC-MS/MS and TNF-α by ELISA. Routine biochemical parameters were evaluated using an automatic biochemical analyzer. Correlations were estimated by Spearman or Pearson test. Statistical significance was set at the level of p < 0.05. Results: All patients presented TMAO levels within the normal range according to EUTox (normal range: 2.83 ± 1.53 mg/L; CAD patients: 0.2 [0.1 to 0.2] ng/μL). TMAO plasma levels were positively correlated with NF-κB mRNA expression (0.555; p = 0.002). Conclusion: TMAO plasma levels may be associated with NF-κB mRNA expression in patients with CAD and may contribute to the pathogenesis of this disease.

El factor de transcripción nuclear NF-κB en cáncer

El factor nuclear κB (NF-κB) es una familia de factores de transcripción sumamente importantes que regulan una gran variedad de genes en diferentes procesos de las respuestas inmunitarias e inflamatorias. Esta familia está compuesta por cinco miembros relacionados estructuralmente, y pueden inducir la transcripción de genes diana al unirse a segmentos específicos de ácido desoxirribonucleico (ADN). Las proteínas NF-κB son usualmente secuestradas en el citoplasma por una familia de proteínas inhibidoras, sin embargo, diversas vías de señalización oncogénica pueden activarla y desencadenar fenotipos malignos en las células correspondientes. El objetivo principal de esta revisión es comprender los mecanismos de regulación del factor de transducción NF-κB, su patogénesis y sus posibles blancos terapéuticos en cáncer. Se consultaron diferentes bases de datos que incluyeron PubMed, Scopus y SciELO, desde el año 2000 hasta diciembre del año 2022; se buscaron las referencias bibliográficas en relación con las palabras clave asociadas al factor NF-κB y cáncer, para finalmente desarrollar la revisión. El factor nuclear de transcripción NF-κB es importante en muchas vías de señalización celular, participa en diversos procesos biológicos y sus alteraciones están asociadas a trastornos inmunitarios y cáncer, entre otras patologías. NF-κB se expresa en todos los tipos de células y tejidos, de tal forma que muchas mutaciones oncogénicas contribuyen a la activación de NF-κB en las células tumorales, y son nuevas rutas de investigación terapéuticas para el cáncer. Existen dos vías de señalización diferentes de NF-κB, denominadas vía canónica y la vía alternativa (no canónica), con distintos mecanismos de activación. Los mecanismos oncogénicos en las que participa el factor NF-κB incluyen inflamación crónica, proliferación, apoptosis, angiogénesis, acción sobre células madre del cáncer, metástasis, regulación metabólica y otros mecanismos asociados. En conclusión, existen aún muchas incógnitas sobre los mecanismos y funciones de NF-κB en el contexto celular; el bloqueo completo del factor NF-κB no parece ser una estrategia factible para el tratamiento del cáncer en el momento actual por la diversidad de acciones fisiológicas importantes que se alteran ante su bloqueo. Futuras investigaciones del factor nuclear NF-κB deberían centrarse en la inhibición de la actividad promotora del cáncer, evitando afectar sus funciones fisiológicas normales.

The nuclear factor kappa B (NF-κB) family of transcription factors, which regulates a large range of genes in various immunological and inflammatory response pathways, is of utmost importance. This family consists of five structurally similar members that can activate target genes by attaching to particular regions of deoxyribonucleic acid (DNA). A class of inhibitory proteins usually keep NF-κB proteins in the cytoplasm; however, different oncogenic signaling pathways can activate them and cause malignant phenotypes in the appropriate cells. The main goal of this review article is to understand the regulatory mechanisms of NF-κB transcription factor, its pathogenesis and its potential cancer therapies. From the year 2000 to December 2022, several databases, including PubMed, Scopus and SciELO, were consulted. Finally, the review was developed by searching bibliographic references looking for the keywords related to NF-κB and cancer. The NF-κB transcription factor plays a key role in numerous cell signaling pathways, is involved in a number of biological functions, and its mutations have been linked to cancer and immunological disorders, among other pathologies. Since NF-κB is expressed in all cell types and tissues, many oncogenic mutations can activate NF-κB in tumor cells, opening up new research possibilities for the treatment of cancer. The canonical pathway and the alternative (non-canonical) pathway are two distinct NF-κB signaling pathways with various activation methods. NF-κB is involved in a variety of oncogenic pathways, including chronic inflammation, proliferation, apoptosis, angiogenesis, effect on cancer stem cells, metastasis, metabolic control and other related mechanisms. In conclusion, there are still many unanswered questions regarding the mechanisms and functions of NF-κB in the cellular context. A complete blockade of NF-κB does not appear to be a feasible strategy for the treatment of cancer at this time due to the variety of significant physiological actions that are altered by its blockade. Future research on NF-κB should focus on preventing cancer promotion while preserving the body's natural physiological processes.

WD repeat domain 5 promotes the development of late-onset preeclampsia by activating nuclear factor kappa B

Purpose: Over-activation of nuclear factor kappa B (NF-κB) was proven to be involved in the pathogenesis of preeclampsia. However, its regulation mechanism is not clear yet. This paper explored the role of WD repeat domain 5 (WDR5) in the development of late-onset preeclampsia and its relationship with NF-κB. Methods: WDR5 expression was detected in normal placentas and placentas from late-onset preeclampsia patients. CCK-8 and colony formation assays were conducted to appraise the proliferative ability of trophoblast. Migration and invasion were observed by wound healing and transwell assays. The interaction between WDR5 and NF-κB inhibitor I-kappa-B-alpha (IkBa) was verified by Co-immunoprecipitation analysis. Immunofluorescence was used to analyze the activation of NF-κB. Finally, we tested the role of WDR5 using the mice late-onset preeclampsia model. Results: WDR5 was highly expressed in the placentas of late-onset preeclampsia patients. WDR5 overexpression suppressed cell proliferation, migration, and invasion in trophoblast. WDR5 could interact with IkBa to activate NF-κB. Knockdown of NF-κB counteracted the anti-proliferative and anti-metastatic effects of WDR5 overexpression in trophoblast. In-vivo studies suggested that targeting WDR5 combated late-onset preeclampsia development. Conclusions: Our finding provides new insights into the role of WDR5 in late-onset preeclampsia development.

The role of TNF-α and NFkß in an experimental model of intestinal carcinogenesis with 1, 2-dimethyhydrazine

ABSTRACT Purpose: To analyze the potential of tumor necrosis factor-α (TNF-α) and factor nuclear kappa B (NF-κB) as colorectal cancer (CRC) biomarkers in an experimental model of intestinal carcinogenesis with 1,2-dimethyhydrazine (1,2-DMH). Methods: Twenty-four male Wistar rats were divided into two groups: sham and 1,2-DMH. First, 1,2-DMH (20 mg/kg/week) was administered for 15 consecutive weeks. In the 25th week, proctocolectomy was conducted. Histopathological analysis, immunohistochemistry, and gene expression of TNF-α and NF-κB were performed. Statistical analysis was performed using GraphPad Prism. The location of aberrant crypt foci (ACF) was analyzed by Kruskal-Wallis' test. For analyses with two groups with parametric data, the t-test was used; for non-parametric data, the Mann-Whitney's test was used. P < 0.05 was considered significant. Results: The number of ACF and macroscopic lesions was significantly higher (p < 0.5) in the 1,2-DMH group compared to the sham group, and most ACF were concentrated in the distal segment of the colon. There was a statistically significant increase (p < 0.5) in protein and gene expression of TNF-α and NF-κB in the 1,2-DMH group compared to the sham group. Conclusions: Our results provide supportive evidence that TNF-α and NF-κB pathways are strongly involved in CRC development in rats and might be used as early biomarkers of CRC pathogenesis in experimental studies.

The effect of SIRT1 activator on inflammatory response in chronic Periodontal disease mice through TLR4/NF-κB signaling pathway / 中国医师杂志

Objective:To explore the effect of silent information regulator 1 (SIRT1) activator SRT1720 on inflammatory response in chronic periodontal disease mice and whether its mechanism is related to the toll like receptor 4 (TLR4)/nuclear factor κB (NF-κB) signaling pathway.Methods:Forty 8-week-old male C57BL/6 mice were selected and divided into a blank control group ( n=8) and an experimental group ( n=32). The experimental group mice were ligated with periodontal pockets and fed with high sugar drinking water. The experimental group was randomly divided into a model group ( n=8) and an SRT1720 group ( n=24). The blank control group and the model group were given physiological saline orally every day. The SRT1720 group was further divided into a low dose group [20 mg/(kg·d), n=8], a medium dose group [50 mg/(kg·d), n=8], and a high dose group [100 mg/(kg·d), n=8] based on the different doses of SRT1720. Four weeks later, the expression levels of interleukin-6 (IL-6), interleukin-1β (IL-1β) and monocyte chemoattractant protein-1 (MCP-1) in gingival crevicular fluid of mice in each group were detected by enzyme linked immunosorbent assay (ELISA); The real-time quantitative polymerase chain reaction (RT-qPCR) method was used to detect the mRNA expression levels of IL-6, IL-1β, MCP-1, SIRT1, TLR4, NF-kB p65 in the gingival tissue of mice in each group; Western blot was used to determine the expression levels of SIRT1, TLR4, and NF-κB p65 proteins in mouse gingival tissue. Results:Compared with the blank control group, the expression levels of IL-6, IL-1β, and MCP-1 inflammatory factors in the gingival crevicular fluid of experimental group mice increased, while the expression levels of IL-6, IL-1β, MCP-1, TLR4, NF-κB p65 mRNA in gingival tissue increased. The expression levels of TLR4, NF-κB p65 protein in gingival tissue increased, while the expression levels of SIRT1 mRNA and protein in gingival tissue decreased, with statistical significance (all P<0.05). Compared with the model group, the expression levels of IL-6, IL-1β, and MCP-1 inflammatory factors in the gingival crevicular fluid, IL-6, IL-1β, MCP-1, TLR4, NF-κB p65 mRNA expression levels in gingival tissue, and TLR4, NF-κB p65 protein expression levels in the gingival tissue of SRT1720 group mice showed a dose-dependent decrease. The expression levels of SIRT1 mRNA and protein in gingival tissue showed a dose-dependent increase, and the differences were statistically significant (all P<0.05). Conclusions:SIRT1 activator SRT1720 can improve the inflammatory response of chronic periodontal disease mice, which may be related to the inhibition of TLR4/NF-kB signaling pathway.

Effects of acupuncture at Houxi(SI3)and Huantiao(GB30)on NF-κB/iNOS/NO pathway in lumbar disc herniation model rats / 针灸推拿医学（英文版）

Objective:To observe the effects of acupuncture at Houxi(SI3)and Huantiao(GB30)on the expression levels of nuclear factor kappa B(NF-κB),inducible nitric oxide synthase(iNOS),and nitric oxide(NO)of NF-κB inflammatory signaling pathway in L5 spinal nerve root of lumbar disc herniation(LDH)model rats and explore the mechanism of acupuncture in LDH treatment.Methods:Forty specific-pathogen-free healthy male Sprague-Dawley rats were randomly divided into a sham operation group,a model group,acupuncture group 1,and acupuncture group 2,with 10 rats in each group.The non-compression nucleus protrusion model was made by puncturing L4-L5 spinous process space and injecting autologous nucleus suspension.Acupuncture at bilateral Shenshu(BL23),Dachangshu(BL25),and Weizhong(BL40)was carried out in acupuncture group 1,and acupuncture at bilateral Houxi(SI3)and Huantiao(GB30)in acupuncture group 2.All rats were treated with balanced reinforcing and reducing needling manipulations,and the needles were retained for 30 min/time with one episode of needling manipulation every 10 min,once a day,14 times in total.The threshold value of paw withdrawal pain was measured by a thermal stimulation pain instrument;the serum NF-κB,iNOS,and NO levels were measured by enzyme-linked immunosorbent assay.The pathomorphological changes of spinal nerve roots were observed by hematoxylin-eosin(HE)staining;quantitative reverse transcription polymerase chain reaction was used to detect iNOS mRNA expression in spinal nerve roots;the NF-κB and iNOS protein expression in spinal nerve roots was detected by the immunofluorescence method.Results:Compared with the sham operation group,the threshold of paw withdrawal pain in the model group was decreased,and the expression levels of serum NF-κB,iNOS,and NO were increased;HE staining showed many degenerated and dissolved Schwann cells in spinal nerve roots with vacuolar changes;meanwhile,the expression levels of NF-κB and iNOS proteins,and the iNOS mRNA in spinal nerve roots were increased.Compared with the model group,the paw withdrawal pain thresholds in acupuncture group 1 and acupuncture group 2 were increased,and the increase in acupuncture group 2 was greater(P<0.05);the expression levels of serum NF-κB,iNOS,and NO in acupuncture group 1 and acupuncture group 2 were decreased,especially in acupuncture group 2(P<0.01);the vacuolar changes of spinal nerve roots,and the degeneration and lysis of Schwann cells in acupuncture group 1 and acupuncture group 2 were decreased,which were more obvious in acupuncture group 2;the NF-κB and iNOS protein expression and the iNOS mRNA expression levels in spinal nerve roots of acupuncture group 1 and acupuncture group 2 were decreased,especially in acupuncture group 2(P<0.01).Conclusion:Acupuncture at Houxi(SI3)and Huantiao(GB30)can improve the morphology of spinal nerve roots,inhibit the NF-κB and iNOS protein expression levels in spinal nerve roots and the serum NO level,and relieve the pain caused by inflammation of spinal nerve roots,which may be one of the mechanisms of acupuncture in LDH treatment.

Role of cold-inducible RNA-binding protein in acute renal injury in a mouse model of myocardial ischemia-reperfusion: relationship with NF-κB signaling pathway / 中华麻醉学杂志

Objective:To evaluate the role of cold-inducible RNA-binding protein (CIRP) in acute renal injury in a mouse model of myocardial ischemia-reperfusion (I/R) and the relationship with nuclear factor kappa B (NF-κB) signaling pathway.Methods:Twenty-four SPF-grade healthy male C57BL/6 mice, aged 6-8 weeks, with body mass index of 24-28 g, were divided into 3 groups ( n=8 each) using a random number table method: sham operation group (Sham group), myocardial I/R group (I/R group) and myocardial I/R + CIRP-derived peptide C23 group (I/R+ C23 group). The model of myocardial I/R was developed by ligation of the left anterior descending coronary artery for 30 min followed by 120-min reperfusion in anesthetized animals. CIRP-derived peptide C23 8 mg/kg was intraperitoneally injected before myocardial ischemia and reperfusion in I/R+ C23 group, while Sham group was only threaded without ligation. Blood samples were collected from the right internal carotid artery at 120 min of reperfusion for determination of the serum creatine kinase isoenzymes (CK-MB), lactic dehydrogenase (LDH), creatinine (Cr) and blood urea nitrogen (BUN) concentrations. Renal tissues were obtained for examination of the pathological changes, and the tubular injury score was assessed. The expression of NF-κB, phosphorylated NF-κB (p-NF-κB), Nod-like receptor protein 3 (NLRP3), interleukin-1beta (IL-1β) and IL-18 in renal tissues was detected by Western blot. The expression of Toll-like receptor 4 (TLR4), NLRP3, IL-1β, TNF-α and IL-6 mRNA was determined by real-time polymerase chain reaction. Results:Compared with Sham group, the levels of serum CK-MB, LDH, Cr and BUN and renal tubule injury score were significantly increased, the expression of p-NF-κB, NLRP3, IL-1β and IL-18 was up-regulated, the expression of TLR4, NLRP3, IL-1β, TNF-α and IL-6 mRNA was up-regulated ( P<0.05), and the pathological injury to renal tissues was aggravated in I/R group. Compared with I/R group, the serum CK-MB, LDH, Cr, BUN and renal tubular injury score were significantly decreased, and the expression of p-NF-κB, NLRP3, IL-1β and and IL-18 was down-regulated, the expression of TLR4, NLRP3, IL-1β, TNF-α and IL-6 mRNA was down-regulated ( P<0.05), and the pathological injury to renal tissues was alleviated in I/R+ C23 group. Conclusions:CIRP is involved in the process of acute renal injury in a mouse model of myocardial I/R, which is associated with activation of NF-κB signaling pathway and promotion of inflammatory responses.

Effect of exosomes derived from bone mesenchymal stem cells on postoperative cognitive function and SIRT1/NF-κB signaling pathway in aged mice / 中华麻醉学杂志

Objective:To evaluate the effect of exosomes derived from bone mesenchymal stem cells (BMSCs-EXO) on the postoperative cognitive function and silent infomation regulator 1 (SIRT1)/ nuclear factor kappa B (NF-κB) signaling pathway in aged mice.Methods:BMSCs-EXO were isolated by differential centrifugation method and then identified. Twenty healthy male C57BL/6 aged mice, aged 18 months, weighing 35-40 g, were divided into 4 groups ( n=5 each) using a random number table method: sham operation group (Sham group), operation group (O group), BMSCs-EXO group and EX527 (SIRT1 inhibitor)group. The abdomen regions were shaved for sterilization without exploratory laparotomy in Sham group. Exploratory laparotomy was performed in O group. BMSCs-EXO 50 μg was injected through the tail vein at 1 h before surgery in BMSCs-EXO group. EX527 5 mg/kg was intraperitoneally injected daily at 1-3 days before surgery, and BMSCs-EXO 50 μg was injected through the tail vein at 1 h before surgery in EX527 group. Morris water maze test was used to evaluate the learning and memory ability for 5 consecutive days staring from the 1st day after surgery. Mice were sacrificed at 1 h after the end of Morris water maze test on day 5 after surgery, and the hippocampal tissues were collected for observation of the pathological changes of hippocampal CA1 region and for determination of the expression of tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6) and IL-1β mRNA (quantitative real-time polymerase chain reaction) and SIRT1 and NF-κB p65 (by Western blot). Results:Compared with Sham group, the escape latency was significantly prolonged, the times of original platform crossing were decreased, the swimming time spent in the original platform quadrant was shortened, the expression of tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6) and IL-1β mRNA was up-regulated, the SIRT1 expression was down-regulated, the NF-κB p65 expression was up-regulated ( P<0.05), and the pathological changes of hippocampal tissues in CA1 region were found in O group. Compared with O group, the escape latency was significantly shortened, the times of original platform crossing were increased, the swimming time spent in the original platform quadrant was prolonged, the expression of TNF-α, IL-6 and IL-1β mRNA was down-regulated, the expression of SIRT1 was up-regulated, the expression of NF-κB p65 was down-regulated ( P<0.05), and the pathological changes of hippocampal tissues in CA1 region were significantly attenuated in BMSCs-EXO group ( P<0.05). Compared with BMSCs-EXO group, the escape latency was significantly prolonged, the times of original platform crossing were decreased, the swimming time spent in the original platform quadrant was shortened, the expression of TNF-α, IL-6 and IL-1β mRNA was up-regulated, the SIRT1 expression was down-regulated, the NF-κB p65 expression was up-regulated ( P<0.05), and the pathological changes of hippocampal tissues in CA1 region were accentuated in EX527 group. Conclusions:BMSCs-EXO can improve the postoperative cognitive function in aged mice, and the mechanism may be associated with the activation of SIRT1/NF-κB signaling pathway.

Role of TLR4/NF-κB signaling pathway in long-term cognitive impairment induced by multiple exposures to sevoflurane anesthesia in neonatal rats / 中华麻醉学杂志

Objective:To evaluate the role of Toll-like receptor 4 (TLR4)/nuclear transcription factor κB (NF-κB) signaling pathway in long-term cognitive impairment induced by multiple exposures to sevoflurane anesthesia in neonatal rats.Methods:Seventy-five SPF healthy newborn Sprague-Dawley rats of either sex, aged 6 days, weighing 12-20 g, were divided into 3 groups ( n=25 each) using a random number table method: control group (group C), multiple exposures to sevoflurane anesthesia group (group S) and TLR4 inhibitor plus multiple exposures to sevoflurane anesthesia group (group I+ S). The rats in group S and group I inhaled 3% sevoflurane for 2 h at 6, 7 and 8 days after birth. TLR4 inhibitor TAK-242 10 mg/kg was intraperitoneally injected before each exposure to sevoflurane in group I, and the equal volume of normal saline was given instead in the other two groups. The spontaneous activity was evaluated by open field test on day 29 after birth, and the cognitive function was assessed by Morris water maze test on days 30-34 after birth. After the behavioral test, the blood samples from the abdominal aorta were collected, and then the rats were sacrificed under deep anesthesia to isolate the hippocampal tissues for measurement of the levels of S100β and neuron-specific enolase (NSE) in serum and hippocampal interleukin-1β (IL-1β), IL-6 and tumor necrosis factor α (TNF-α) (by enzyme-linked immunosorbent assay), expression of TLR4, NF-κB p65 and phosphorylated NF-κB p65 (p-NF-κB p65) (by Western blot) and for microscopic examination of the pathological changes of hippocampal CA1 region after HE staining. Results:Compared with group C, the escape latency was significantly prolonged, the number of crossing the original platform was reduced, the TLR4 expression was up-regulated, the ratio of p-NF-κB p65/NF-κB p65 was increased, the levels of serum S100β protein and NSE and hippocampal IL-1β, IL-6 and TNF-α were increased ( P<0.05), and the pathological changes in the hippocampal CA1 region were aggravated in group S. Compared with group S, the escape latency was significantly shortened, the number of crossing the original platform was increased, TLR4 expression was down-regulated, the ratio of p-NF-κB p65/NF-κB p65 was decreased, the levels of S100β and NSE in serum and hippocampal IL-1β, IL-6 and TNF-α were decreased ( P<0.05), and the pathological changes in hippocampal CA1 area were significantly attenuated in group P. Conclusions:The mechanism by which multiple exposures to sevoflurane anesthesia induces long-term cognitive impairment is related to activation of TLR4/NF-κB signaling pathway and increase in hippocampal inflammatory responses in neonatal rats.

Role of SIRT1/NF-κB signaling pathway in mild hypothermia-induced promotion of microglial polarization during oxygen-glucose deprivation and restoration / 中华麻醉学杂志

Objective:To evaluate the role of Sirtuin 1/nuclear factor κB (SIRT1/NF-κB) signaling pathway in mild hypothermia-induced promotion of microglial polarization during oxygen-glucose deprivation and restoration (OGD/R).Methods:The well-grown BV2 microglia were divided into 4 groups ( n=36 each) using the random number table method: control group (group C), OGD/R group (group O), mild hypothermia group (group M), and mild hypothermia+ SIRT1 specific inhibitor EX527 group (group ME). Cells in group C were commonly cultured without any treatment. Cells in group O were subjected to 3 h of OGD followed by 21 h of restoration of O 2-glucose supply at 37 ℃. Cells in group M were subjected to 3 h of OGD followed by 21 h of restoration of O 2-glucose supply at 33 ℃. Cells in group ME were co-cultured with inhibitor EX527 (final concentration 5 nmol/L) for 12 h in the medium before OGD/R, and the other procedures were conducted as previously described in group M. The cell survival rate was detected by CCK-8 assay. The levels of tumor necrosis factor-alpha (TNF-α), interleukin-1beta (IL-1β) and interleukin-10 (IL-10) in supernatant were detected by enzyme-linked immunosorbent assay. The expression of CD206, CD32, inducible nitric oxide synthase (iNOS) and arginine synthase 1 (Arg-1) mRNA was detected by quantitative real-time polymerase chain reaction. The expression of CD206 and CD32 was detected by immunofluorescent staining. The expression of iNOS, Arg-1, SIRT1, NF-κB p65 (p65) and acetylated NF-κB (Ac-p65) was detected by Western blot. Results:Compared with group C, the cell survival rate was significantly decreased, the concentrations of TNF-α, IL-6 and IL-10 in the supernatant were increased, the expression of CD206, Arg-1, CD32 and iNOS was up-regulated, the expression of SIRT1 was down-regulated, and the Ac-p65/p65 ratio was increased in group O ( P<0.05). Compared with group O, the cell survival rate was significantly increased, the concentrations of TNF-α and IL-6 in the supernatant were decreased, the concentration of IL-10 was increased, the expression of CD206, Arg-1 and SIRT1 was up-regulated, the expression of CD32 and iNOS was down-regulated, and the Ac-p65/p65 ratio was decreased in group M ( P<0.05). Compared with group M, the cell survival rate was significantly decreased, the concentrations of TNF-α and IL-6 in the supernatant were increased, the concentration of IL-10 was decreased, the expression of CD206, Arg-1 and SIRT1 was down-regulated, the expression of CD32 and iNOS was up-regulated, and the Ac-p65/p65 ratio was increased in group ME ( P<0.05). Conclusions:SIRT1/NF-κB signaling pathway is involved in mild hypothermia-induced promotion of microglial polarization during OGD/R.

Research advances in the cascade interaction between reactive oxygen species/reactive nitrogen species and the NF-κB signaling pathway in liver fibrosis / 临床肝胆病杂志

Liver fibrosis is a compensatory response in the process of tissue repair after chronic liver injury, and it is also a necessary pathological process in the progression of a variety of chronic liver diseases. In the pathological state, the imbalance between hepatic oxidative system and antioxidant system can lead to the excessive production or insufficient clearance of reactive oxygen species (ROS)/reactive nitrogen species (RNS), which may induce the injury of hepatocytes, expand inflammatory response, and promote the development and progression of liver fibrosis. As a master regulator of oxidative stress and inflammatory response, NF-κB plays a key role in the process of liver fibrosis. Therefore, the cascade interaction between ROS/RNS and the NF-κB signaling pathway plays a guiding role in further clarifying the pathogenesis of liver fibrosis and exploring effective prevention and treatment strategies. This article reviews and discusses the interaction between ROS/RNS and the NF-κB signaling pathway and its important role in the progression of liver fibrosis, so as to provide strategies and references for targeted therapy for liver fibrosis.

The protective effect and mechanism of Ophiopogonin D on LPS-induced mouse macrophages / 国际中医中药杂志

Objective:To study the protective effect of Ophiopogonin D on lipopolysaccharide (LPS)-induced mouse macrophage RAW264.7 and its related mechanism.Methods:Mouse macrophage RAW264.7 cells were cultured and divided into normal control group, model group and Ophiopogonin D pretreatment group according to random number table method. The activity of Ophiopogonin D on RAW264.7 cells was detected by CCK-8 method; the protein levels of TNF-α, IL-1β, IL-6, reactive oxygen species (ROS), MDA and SOD were detected by ELISA; the protein expression of NF-κB, TLR4, NF-E2-related factor2 (Nrf2) and heme oxygenase-1 (HO-1) were detected by Western Blot.Results:Compared with model group, the levels of TNF-α, IL-1β, IL-6, ROS and MDA in cell supernatant of Ophiopogonin D group were decreased ( P<0.05), and the level of SOD was increased ( P<0.05). The protein expressions of NF-κB (0.76±0.10 vs. 2.26±0.17) and TLR4 (0.98±0.09 vs. 1.74±0.19) significantly decreased ( P<0.05). The protein expressions of Nrf2 (0.85±0.03 vs. 0.54±0.03) and HO-1 (0.97±0.11 vs. 0.37±0.04) significantly increased ( P<0.05). Conclusion:Ophiopogonin D may reduce inflammatory response by reducing TLR4/NF-κB pathway, and activate Nrf2/HO-1 pathway to reduce oxidative damage and play a protective role.

Experimental study on the improvement of inflammatory response in rats with cerebral ischemia by Chinese ultrasound drug permeation electrotherapy device / 国际中医中药杂志

Objective:To observe the effects of Traditional Chinese Medicine (TCM)ultrasound drug permeation electrotherapy device on the inflammatory response of rats with cerebral ischemia, and to provide an experimental basis for the clinical application of TCM ultrasound drug permeation electrotherapy device in the treatment of cerebral ischemia.Methods:A total of 72 SD rats were randomly divided into sham-operation group (12 rats) and modeling group (60 rats). The middle cerebral artery occlusion (MCAO) model was prepared by thread embolism in the model group. The rats were divided into model group, Chinese medicine tablet group, blank tablet + TCM ultrasound drug permeation electrotherapy group (hereinafter referred to as "blank tablet + electrotherapy group"), Chinese medicine tablet + TCM ultrasound drug permeation electrotherapy group (hereinafter referred to as "Chinese medicine tablet + electrotherapy group") and butylphthalide group according to the random number table method, with 12 rats in each group. The corresponding treatment was given continuously for 7 days. The neurological function was scored using Longa method evaluation criteria; TTC staining was used to observe the infarct volume and calculate the percentage of infarct volume; HE staining was used to observe the cell morphology of cortical area in each group of rats; ELISA was used to detect the serum TNF-α and IL-1β levels in each group of rats; TLR4, MyD88 and NF-κBp65 protein expressions in hippocampal tissue of each group of rats on the infarct side were detected by Western blot method.Results:Compared with the model group, the neurological function scores of rats in the blank tablet + electrotherapy group, the herbal tablet + electrotherapy group, and the butylphthalein group significantly decreased ( P<0.05), the percentage of cerebral infarct volume significantly decreased ( P<0.05), the contents of serum TNF-α and IL-1β significantly decreased ( P<0.05), and the expressions of TLR4 (0.42±0.07, 0.31±0.07, 0.19±0.04 vs. 0.68±0.14), MyD88 (0.39±0.12, 0.30±0.07, 0.23±0.11 vs. 0.67±0.10), NF-κBp65 (0.32±0.03, 0.27±0.02, 0.17±0.03 vs. 0.57±0.12) protein in hippocampal tissue significantly decreased ( P<0.05). Conclusion:The TCM ultrasound drug permeation electrotherapy device can inhibit TLR4, MyD88, NF-κBp65 protein expressions and reduce the release of serum inflammatory factors TNF-α and IL-1β, thus exerting cerebral ischemic protective effects.

Clinical Significance of HSCARG for Atherosclerotic Coronary Heart Disease and Reduced ROS-Oxidative Stress in in Vivo and in Vitro Models via p47phox by NF-κB Activity

ABSTRACT Introduction: Coronary heart disease (CHD) is a dynamic process in which there are interactions between endothelial dysfunction, oxidative stress, and inflammatory responses. The aim of the present study was to investigate the function and mechanism of HSCARG in the treatment of CHD. Methods: Male apolipoprotein E/low-density lipoprotein receptor-deficient mice were given a high-fat diet with 21% fat and 0.15% cholesterol for the in vivo model. Human umbilical vein endothelial cells were incubated with angiotensin II for the in vitro model. HSCARG expression was inhibited in patients or mice with CHD. Results: HSCARG reduced oxidative stress in mice with CHD. HSCARG also reduced reactive oxygen species (ROS)-oxidative stress in the in vitro model. HSCARG induced p47phox expression in the in vitro model by NF-κB activity. The regulation of nuclear factor kappa B (NF-κB) activity or p47phox expression participates in the effects of HSCARG in CHD. Conclusion: Altogether, our data indicate that HSCARG reduced ROS-oxidative stress in in vivo and in vitro models of CHD via p47phox by NF-κB activity and may be a clinical target for CHD.

Is -94ins/delattg polymorphism in the nuclear factor kappa-b1 gene (nfkb1) associated with necrotizing enterocolitis? / Polimorfismo -94ins/delattg no gene do fator nuclear kappa-b1 (nfkb1) está associado à enterocolite necrosante?

ABSTRACT BACKGROUND: Abnormalities in the different stages of the intestinal maturation process cause metabolic and molecular changes. Among the genetic alterations associated with necrotizing enterocolitis, the -94ins/delATTG polymorphism in NFKB1 gene leads to unregulated activation of the NFKB protein due to an increase in the inherent pro-inflammatory state of the premature intestine. AIMS:To determine the prevalence of the -94ins/delATTG polymorphism in NFKB1 gene in neonates with and without necrotizing enterocolitis. METHODS:This is a case-control study, in which 25 neonates were evaluated as the case group and 50 neonates as the control group, of both genders. DNA was extracted from peripheral blood leukocytes, and the site encompassing the polymorphism was amplified by molecular techniques (polymerase chain reaction/polymorphism in restriction fragment length). RESULTS:Necrotizing enterocolitis was diagnosed in 25 (33%) neonates and, of these, 3 (12%) died. Male gender was more prevalent in both groups (p=0.1613): cases (52%) and controls (62%). Moderate and extreme preterm newborns were predominant in both groups: cases (80%) and controls (88%) (p=0.3036). Low birth weight and extremely low birth weight newborns were the most prevalent in cases (78%), and very low birth weight and extremely low birth weight were the most prevalent in controls (81%) (p=0.1073). Clinical treatment was successful in 72%, and hospital discharge was achieved in 88% of newborns with NEC. The -94ins/delATTG polymorphism in NFKB1 gene was not identified in all the 150 alleles analyzed (100%). CONCLUSIONS:The absence of the -94ins/delATTG polymorphism in NFKB1 gene in newborns with and without necrotizing enterocolitis does not rule out the possibility of alterations in this and/or in other genes in newborns with this condition, which reinforces the need for further research.

RESUMO RACIONAL:Anormalidades nas diferentes fases do processo de maturação intestinal causam alterações metabólicas e moleculares. Dentre as alterações genéticas associadas à enterocolite necrotizante, o polimorfismo -94ins/delATTG no gene NFKB1 leva à ativação desregulada da proteína NFKB devido ao aumento do estado pró-inflamatório inerente ao intestino prematuro. OBJETIVOS:Determinar a prevalência do polimorfismo -94ins/delATTG no gene NFKB1 em neonatos com e sem enterocolite necrotizante. MÉTODOS: Trata-se de um estudo caso-controle, no qual foram avaliados 25 neonatos como grupo caso e 50 neonatos como grupo controle, de ambos os sexos. O DNA foi extraído de leucócitos do sangue periférico e o sítio que engloba o polimorfismo foi amplificado por técnicas moleculares (reação em cadeia da polimerase/polimorfismo no comprimento do fragmento de restrição). RESULTADOS: Enterocolite necrosante foi diagnosticada em 25 (33%) neonatos e, destes, 3 (12%) foram a óbito. O gênero masculino foi mais prevalente em ambos os grupos (p=0,1613): casos (52%) e controles (62%). Os prematuros moderados e extremos foram predominantes em ambos os grupos: casos (80%) e controles (88%) (p=0,3036). Recém-nascidos de baixo peso e extremo baixo peso foram os mais prevalentes nos casos (78%) e de muito baixo peso e extremo baixo peso foram os mais prevalentes nos controles (81%) (p=0,1073). O tratamento clínico foi bem-sucedido em 72% e a alta hospitalar foi obtida em 88% dos recém-nascidos com enterocolite necrotizante. O polimorfismo -94ins/delATTG no gene NFKB1 não foi identificado em todos os 150 alelos analisados (100%). CONCLUSÕES: A ausência do polimorfismo -94ins/delATTG no gene NFKB1 em recém-nascidos com e sem enterocolite necrosante não afasta a possibilidade de alterações neste e/ou em outros genes em recém-nascidos com esta condição, o que reforça a necessidade de novas pesquisas.

Effects of electroacupuncture pretreatment on M1 polarization of alveolar macrophages in rats with acute lung injury / 针灸推拿医学（英文版）

Objective: To observe the effects of electroacupuncture (EA) pretreatment on M1 polarization of alveolar macrophages (AMs) in rats with acute lung injury (ALI) induced by lipopolysaccharide (LPS), and to explore the potential protective mechanism of EA.Methods: Forty Sprague-Dawley rats were randomly divided into a normal group, a model group, and three groups of EA pretreatment [including a Chize (LU5) group, a Zusanli (ST36) group and a Chize (LU5) plus Zusanli (ST36) group], with eight rats in each group. The model rats of ALI were established by instilling LPS [2 mg/(kg·bw)] into the trachea of rats for 3 h. The rats in each EA pretreatment group were pretreated with EA for 30 min per day at the corresponding bilateral acupoints 6 d before instilling LPS. Three hours after modeling, the pulmonary function of the rats was tested, and the lung tissue was taken to calculate the ratio of lung wet weight to dry weight (W/D). The pathological lung changes and the injury score were observed by hematoxylin-eosin staining. The contents of tumor necrosis factor (TNF)-α, interleukin (IL)-1β, and myeloperoxidase (MPO) in rat's bronchoalveolar lavage fluid (BALF) were detected by enzyme-linked immunosorbent assay. The mRNA and protein expression levels of M1 macrophage markers clusters of differentiation 86 (CD86), inducible nitric oxide synthase (iNOS), and its signaling pathway factor Toll-like receptor (TLR) 4, and nuclear factor-κB (NF-κB) p65 in the alveoli were detected by fluorescence quantitative polymerase chain reaction and Western blot, respectively. Results: After being induced by LPS, the pulmonary function of the model rats showed that the forced expiratory volume in 0.1 s (FEV0.1), forced expiratory volume in 0.3 s (FEV0.3), and their respective ratios of FEV to forced vital capacity (FVC) (including FEV0.1/FVC and FEV0.3/FVC) were significantly decreased (P<0.01), while the W/D of lung tissue was increased (P<0.01). The score of lung injury was significantly higher (P<0.01). The contents of TNF-α, IL-1β, and MPO in the BALF and the mRNA and protein expression levels of CD86, iNOS, TLR4, and NF-κB p65 in the lung tissue were significantly increased (P<0.01). After EA pretreatment, the FEV0.1, FEV0.3, FEV0.1/FVC, and FEV0.3/FVC were significantly increased, the lung injury score decreased significantly, and the contents of TNF-α, IL-1β, and MPO in the BALF and the expression levels of CD86, iNOS, TLR4, and NF-κB p65 mRNAs and proteins in the alveoli decreased significantly (P<0.05 or P<0.01). Compared with the other two single acupoint groups, the contents of TNF-α, IL-1β, and MPO in the BALF and the expression levels of CD86, iNOS, TLR4, and NF-κB p65 mRNAs in the alveoli in the Chize (LU5) plus Zusanli (ST36) group were significantly lower (P<0.01). Conclusion: EA pretreatment at Chize (LU5) and Zusanli (ST36) can inhibit inflammation and reduce pulmonary injury in ALI rats induced by LPS. The effect of the combination of Chize (LU5) and Zusanli (ST36) is better than that of using these two acupoints separately, and its mechanism may be related to the inhibition of AMs' M1 polarization by down-regulation TLR4/NF-κB signaling pathway.

Effects of iron metabolism and oxidative stress level on blood glucose control during pregnancy in patients with gestational diabetes mellitus / 中国基层医药

Objective:To investigate the effects of iron metabolism and oxidative stress level on blood glucose control during pregnancy in patients with gestational diabetes mellitus (GDM).Methods:A total of 139 pregnant women who received prenatal examination between January 2020 and June 2021 in Wenzhou Central Hospital were included in this study. They were divided into GDM group ( n = 68) and control group ( n = 71) according to oral glucose tolerance test results at 24-48 weeks of gestation. Clinical data were collected. Iron metabolism, oxidative stress and blood glucose levels were measured. The relationships between iron metabolism and oxidative stress levels and blood glucose control in GDM were analyzed. Results:There was no significant difference in age between the GDM and control groups ( P > 0.05). Body mass index, fasting blood glucose, fasting insulin, glycosylated hemoglobin, nuclear factor-κB (NF-κB), malondialdehyde (MDA), ferritin (SF), serum iron, transferrin (TRF) and insulin resistance index (IRI) in the GDM group were (24.11 ± 3.05) kg/m 2, (4.92 ± 0.67) mmol/L, (10.56 ± 2.21) pmol/mL, (6.15 ± 0.62)%, (20.50 ± 1.72) μg/L, (20.34 ± 2.92) μmol/L, (70.77 ± 7.01) μg/L, (30.18 ± 4.25) μmol/L, (3.93 ± 0.69) g/L and (2.50 ± 1.03), respectively, which were significantly higher than those in the control group [(21.41 ± 2.86) kg/m 2, (4.69 ± 0.62) mmol/L, (5.76 ± 2.09) pmol/mL, (5.37 ± 0.58)%, (15.43 ± 1.55) μg/L, (12.93 ± 2.17) μmol/L, (42.53 ± 8.86) μg/L, (18.81 ± 3.85) μmol/L, (2.89 ± 0.53) g/L and (1.74 ± 0.89)] ( t = 5.39, 2.10, 13.16, 7.66, 18.27, 17.03, 20.78, 16.54, 9.99, 4.66, all P < 0.05). Superoxide dismutase (SOD), total antioxidant capacity (TAOC) and insulin sensitivity index in the GDM group were (21.49 ± 3.52) U/L, (10.87 ± 1.34) kU/L and (3.28 ± 0.46), respectively, which were significantly lower than those in the control group [(26.28 ± 3.95) U/L, (13.28 ± 1.52) kU/L, (3.86 ± 0.53), t = 7.54, 9.90, 6.88, all P < 0.05]. Multivariate logistic regression analysis revealed that SOD, TAOC, NF-κB, MDA, SF and TRF were independent influential factors of GDM occurrence [ OR (95% CI) = 1.57 (1.09-2.26), 3.15 (1.71-5.80), 2.18 (1.32-3.61), 3.27 (1.58-6.76), 2.12 (1.29-3.50), 1.23 (0.99-1.53), 3.65 (1.89-7.04), all P < 0.05]. SOD and TAOC levels were negatively correlated with IRI ( r = -0.75, -0.84, both P < 0.05), while NF-κB, MDA, SF, serum iron and TRF were positively correlated with IRI ( r = 0.93, 0.96, 0.98, 0.07, 0.92, all P < 0.05). Conclusion:Increased levels of iron metabolism and oxidative stress are risk factors for the occurrence of GDM, and they are closely related to the degree of insulin resistance. GDM screening should be carried out in advance in pregnant women with increased levels of iron metabolism and oxidative stress indicators, which plays a positive role in clinical diagnosis and treatment of GDM.

Long chain noncoding RNA FAM224B protects the lung tissue of rats with severe pneumonia and the underlying mechanism / 中国基层医药

Objective:To investigate the protective effects of overexpression of long-chain noncoding RNA FAM224B on lung tissue of rats with severe pneumonia and the underlying mechanism.Methods:From August 2020 to March 2021, we randomly allocated 20 rats into the pneumonia group (severe pneumonia modeling) and FAM224B group (severe pneumonia modeling + FAM224B plasmid), with 10 rats in each group. We performed a quantitative real-time polymerase chain reaction to detect the level of FAM224B in lung tissue and performed an enzyme-linked immunosorbent assay to detect the levels of tumor necrosis factor-alpha, interleukin-6, and interleukin-1β in lung tissue. We used the software starBase v2.0 to predict the target gene of FAM224B. We performed a quantitative real-time polymerase chain reaction to detect the expression of the target gene in lung tissue and performed a western blot assay to detect the protein expression of the nuclear factor-kappa B signal pathway in lung tissue.Results:FAM224B expression was (1.09 ± 0.23) and (10.12 ± 1.52) in the pneumonia and FAM224B groups, respectively. FAM224B expression was significantly lower in the pneumonia group compared with the FAM224B group ( t = 15.86, P < 0.01). The levels of tumor necrosis factor-alpha, interleukin-6, and interleukin-1β were (41.53 ± 2.46) μg/L, (34.01 ± 2.53) ng/L, (20.92 ± 1.95) μg/L in the pneumonia group and they were (21.71 ± 2.25) μg/L, (17.13 ± 3.01) ng/L, (11.97 ± 1.21) μg/L in the FAM224B group. There were significant differences in the levels of tumor necrosis factor-alpha, interleukin-6, and interleukin-1β between the two groups ( t = 15.94, 14.29, 13.89, all P < 0.01). FAM224B had complementary binding sites with miR-34b-5p. The expression level of miR-34b-5p in lung tissue was significantly lower in the FAM224B group compared with the pneumonia group ( t = 15.55, P < 0.01). The protein expression levels of phosphorylated nuclear factor-κB subunit (p-p65) and phosphorylated inhibitor of kappa B alpha in lung tissue were significantly lower in the FAM224B group compared with the pneumonia group. Conclusion:FAM224B overexpression reduces the inflammatory reaction in lung tissue of rats with severe pneumonia through inhibiting miR-34b-5p expression.

Effect of Rehmanniae Radix combined with Scrophulariae Radix on renal microinflammation in diabetic nephropathy rats based on NF-κB pathway / 国际中医中药杂志

Objective:To explore the effect of Rehmanniae Radix combined with Scrophulariae Radix on renal microinflammation in diabetic nephropathy (DN) rats. Methods:50 Sprague Dawley (SD) rats were adaptively fed for 1 week, and then 10 rats were randomly selected as the blank control group, and the rest were treated with STZ intraperitoneal injection combined with high-fat diet to induce DN model. After 4 weeks, the successful modeled rats were randomly divided into model group, Rehmannia glutinosa Scrophularia group (5.25 g/kg) and metformin group (200 mg/kg), with 10 rats in each group. After 8 weeks of administration, fasting blood glucose was measured by blood glucose meter; microalbuminuria was measured by benzalkonium chloride turbidimetry; serum cystatin, TNF-α, IL-6 and hs-CRP levels were measured by ELISA kit; renal pathological changes were detected by HE staining, Masson staining and PAS staining; the expression of MCP-1, NF-κB (total) and p-NF-κB protein in renal tissue was detected by Western blot.Results:Compared with the model group, the body weight of rats in DHXS group was significantly decreased ( P<0.05). The content of fasting blood glucose[(18.06 ± 5.69) mmol/L vs. (29.42 ± 0.63)mmol/L], 24-hour urine protein [(11.02 ± 1.77)mg/d vs. (31.61 ± 0.65)mg/d], serum cystatin [(208.16 ± 12.07)ng/ml vs. (278.05 ± 19.33)ng/ml], TNF-α [(9.13 ± 1.46)pg/ml vs. (73.16 ± 8.30)pg/ml], IL-6[(4.27 ± 1.07)pg/ml], hs-CRP[(219.36 ± 22.02)ng/ml vs. (266.97 ± 15.80)ng/ml] in DHXS group were significantly decreased ( P<0.05), and the expression level of p-NF-κB (0.49 ± 0.07 vs. 0.84 ± 0.12) and MCP-1 (0.44 ± 0.02 vs. 0.64 ± 0.11) in renal tissue of rats in DHXS group were significantly reduced ( P<0.05). Conclusion:Rehmanniae Radix combined with Scrophulariae Radix can protect kidney by inhibiting the over activation of NF-κB, and reducing the expression of MCP-1 related protein to reduce renal micro inflammation.

Research process on pathogenesis of gastroesophageal reflux disease / 中国医师杂志

Gastroesophageal reflux disease (GERD) is a gastrointestinal motility disorder that results from the reflux of stomach contents into the esophagus or oral cavity, causing symptoms or complications. The typical symptoms of GERD are heartburn and regurgitation of gastric contents into the oropharynx. Heartburn is the sensation of burning or discomfort behind the sternum. Heartburn may radiate into the neck, is typically worse after meals or when in a reclining position, and may be eased by antacids. Regurgitation is the backflow of gastric contents into the mouth or hypopharynx. Epigastric pain can also be a symptom of GERD. Extraesophageal symptoms of GERD include dental erosions, laryngitis, cough, and asthma. In recent years, great progress has been made in understanding the molecular basis of GERD, suggesting that its pathogenesis is more complex and multifactorial. In this paper, the molecular pathogenesis was taken as the starting point, including the mechanism of genes in the pathogenesis and development of GERD, the mechanism of NF-κB pathway in the pathogenesis and development of GERD, the role of proteinase-activated receptor-2 in the pathogenesis of GERD, the association between abnormal serotonin pathway and GERD, and the relationship between reactive oxygen species and GERD, to summarize the pathogenesis of gastroesophageal reflux disease.