Expression Levels Of NF-kB Research Articles

A study on the effects of replacing fishmeal with soybean meal in the feed of Procambarus clarkii: Assessing growth performance, immunity, and gut microbiota

The unstable supply and high cost of fishmeal have led to increased interest in low fish meal feed. As such, the present study aimed to investigate the effects of replacing fishmeal in feeds with soybean meal (SM) on the growth, immunity, and gut microbiota of crayfish. A total of 750 crayfish (4.00 g) were randomly divided into five groups (SS0, SS25, SS50, SS75, and SS100) with three replicates, and were fed diets with different levels of soybean meal substituted for fishmeal for six weeks. The results revealed that crayfish subjected to high SM diets experienced a significant decrease in the final body weight, weight gain rate, and specific growth rate (p<0.05), together with enzyme activities related to nutrient digestion and absorption. Of note, the groups with 25–50 % SM replacement exhibited decreased final body weight without significance compared to that in the SS0 group (p>0.05). Moreover, increasing SM inclusion exhibited the potential to impair the intestine, characterized by the increase in hemolymph LD content, decrease in the levels of TC content, T-AOC, and AKP, along with the observed edema of the lamina propria, and up-regulated mRNA expression levels of nfkb, alf, and tlr. Furthermore, high SM diets resulted in the emergence of new species and a shift in Firmicutes, Bacteroidota, Proteobacteria, and Fusobacteriota. Specifically, the abundance of lower Firmicutes and higher Proteobacteria was observed in the SS100 group, which was closely correlated with genus-level Candidatus Bacilloplasma, and RsaHf231 abundance alterations. MENAs demonstrated that the high SM diets led to a more complex, yet fragile and unstable microbial network. The global network further indicated that the microbes contributing more to crayfish were in module eigen 4, in which 9 OTUs were identified as major biomarkers and mainly classified into Proteobacteria. Taken above, SM could replace 25 % of fishmeal in crayfish diets without negatively affecting growth performance, nutrient utilization, immunity, and antioxidant capacity. Excessive SM levels contributed to gut dysfunction and weakened the immune system of crayfish.

Inhibition of NF-kB and COX-2 by andrographolide regulates the progression of cervical cancer by promoting PTEN expression and suppressing PI3K/AKT signalling pathway

In the face of recent advances in Cervical cancer (CC) treatment, therapeutic and surgical procedures for CC management are still inadequate. In the current study for the first time Andrographolide (Andro) has been explored for its multitarget therapeutic efficacy on NF-kB, COX-2, and PI3K/AKT expressions together in CC. The expression levels of NF-kB, COX-2, PI3K and PTEN in the CC patient samples, both at mRNA and protein levels have shown significant association with poor survival and increased tumor aggressiveness. The binding efficacy of Andro was investigated using molecular docking and molecular dynamic simulations, and the protein and ligand complex for NF-kB and COX-2 has shown high binding energy. Andro displayed cytotoxicity by impeding the in-vitro proliferation of CC cells. Andro significantly supressed the NF-kB, COX-2, and PI3K expression and enhanced the expression levels of PTEN at protein levels in-vitro. Andro induced apoptosis in a dose dependent manner and significantly inhibited the migration and invasion of CC cells. Andro exhibited similar activity in-vivo and suppressed the CC tumor growth in xenograft C57BL/6 mice model. The anti-tumor activity of Andro, both in-vitro and in-vivo has shown considerable downregulation of NF-kB and COX-2 and induced apoptosis through impeding the PI3K/AKT signalling pathway. These findings from the above study projects, administration of Andro as an effective alternate safe compound to curtail and impede cervical cancer progression.

#1749 Influence of high saline diet on myocardial remodeling, microRNA-21i and NFkB expression in myocardium

Abstract Background and Aims There is now increased interest in the study of morpho-functional myocardial changes in excess NaCl consumption, which develop independently of AD dynamics. Myocardial remodeling may involve microRNAs that modulate gene expression at the post-transcription level. However, their role in this process has not been fully explored. The purpose of the study is to assess the levels of expression of the nucleic transcription factor kV (NFkB), microRNA-21 and structural changes in the myocardium at long-term consumption of Wistar diet with high (8%) table salt content. Method Male Wistar rats (body mass 280.5 42.7 g) were divided into two groups of 10 animals. For 4 months, the control rats (NaCl) received a standard diet (0.34% NaCl), and the animals of the other group received a high salt diet (8% NaCl) (HS). Laboratory food varied only in NaCl content. 4 months later in rats systolic AD was measured, myocardial mass index (IMM) was evaluated, myocardial histology was performed (paraffin coloration - hematoxylin-eosin, Masson), relative expression levels of my RNA-determined 21 and NFkB in the myocardium, sodium content in the urine and blood serum. Statistical analysis used the t-Student criterion and the Mann-Whitney test. The differences were considered significant at p &amp;lt; 0.05. Results Excess NaCl (8%) in the diet had no effect on animal body weight (370.5 32.0 g in HS group, 393.0 21.4 g in NS group, p &amp;gt; 0.05) and systolic AD. In the HS-Group, the average AD was 134.5 8.9 mm old, and in the NS-Group - 134.8 5.2 mm old. (p &amp;gt; 0.05). However, HS-group IMM rats were 25% higher than NS-group rats (p &amp;lt; 0.05). The daily water consumption in the HS group was on average 33% higher than in the control, the daily diuresis also increased (10.5 5.7 ml against 5.7 2.9 ml in the NS-group, p &amp;lt; 0.001) and the sodium content in the urine (81.25.72 mmol/l against 161.18.57 mmol in the NS-01). There was no difference in the sodium content of the blood serum. Hypertrophy of cardiomyocytes, protein dystrophy, and increased wall thickness of arterial vessels (NS-20,42 3,53, HS-27,61 4,93, µm were identified in the HS-group; p &amp;lt; 0.01). The area of perivascular fibrosis in rats of the HS group (14876 ± 3803.7 µm2) was almost 1.8 times higher than that in the NS group (8426.0 ± 3012.2 µm2, p &amp;lt; 0.03). The thickness of cardiomyocytes (NS-12.97 1.53, HS-20.28 1.90 µm, p &amp;lt; 0.001) also increased in rats on the HS diet. In animals of the HS group, the relative expression levels of NFkB (more than 2 times) and microRNA-21 (almost 6 times) in the myocardium increased compared to the NS group. It can be assumed that the adverse effect on the cardiovascular system of high-salt diets is partially realized through NFκB-associated signaling pathways and the activation of microRNA-21. Conclusion Long-term use of a high-salt diet in Wistar rats results in myocardial remodeling unrelated to changes in AD levels. In this case, the high salt intake effects on the myocardium may be mediated by an increase in NFkBy and microRNA-21 expression levels.

Garden Cress Seed Oil Abrogates Testicular Oxidative Injury and NF-kB-Mediated Inflammation in Diabetic Mice.

Diabetes mellitus is a metabolic disorder associated with various complications encompassing male reproductive dysfunction. The present study aimed to investigate the therapeutic potential of biologically active Lepidium sativum seed oil (LSO) against the testicular dysfunction associated with streptozotocin (STZ)-induced diabetes. Male adults (n = 24) were divided into four groups: control, LSO-administered, diabetic (D), and LSO-treated diabetic (D+LSO) groups. LSO was extracted from L. sativum seeds, and its chemical composition was determined using GC-MS. Serum testosterone levels, testicular enzymatic antioxidants (catalase (CAT) and superoxide dismutase (SOD)), an oxidative stress (OS) biomarker, malondialdehyde (MDA), pro-inflammatory markers (NF-kB, IL-1, IL-6, and TNF-α), and the expression level of NF-kB were assessed. In addition, histopathological changes were evaluated in testicular tissues. The results obtained showed that the chemical composition of LSO indicated its enrichment mainly with γ-tocopherol (62.1%), followed by 2-methylhexacosane (8.12%), butylated hydroxytoluene (8.04%), 10-Methylnonadecane (4.81%), and δ-tocopherol (3.91%). Moreover, LSO administration in the D+LSO mice significantly increased testosterone levels and ameliorated the observed testicular oxidative damage, inflammatory response, and reduced NF-kB expression compared to the diabetic mice. Biochemical and molecular analyses confirmed the histological results. In conclusion, LSO may prevent the progression of diabetes-induced impairment in the testes through inhibition of the OS- and NF-kB-mediated inflammatory response.

Effects of glycation end-products on the dental pulp inpatients with type 2 diabetes.

This ex vivo study aimed to compare protein expression of advanced glycation end-products (AGE) and receptor (RAGE), and the levels of selected genes associated with inflammation and collagen within dental pulp tissue from patients with type 2 (T2D) diabetes and non-T2D. Noncarious extracted permanent molar teeth from patients with well-controlled T2D (n = 19) and non-T2D (controls) (n = 19) were collected and compared. The coronal pulp was examined using immunohistochemistry (IHC) (n = 10 per group) for anti-AGE and anti-RAGE. Quantitative PCR (n = 9 per group) was used to analyse the gene expression levels of NFKB, S100A12 and COLIA1. Data analyses were performed between the groups using GraphPad Prism using Pearson correlation, Shapiro-Wilk and Mann-Whitney U-tests, and multiple regression using SPSS. AGEs were distributed diffusely throughout the pulp extracellular matrix associated with collagen fibres and were present on several cell types. RAGE was expressed at the pulp-dentine interface and was observed on odontoblasts, immune cells, endothelial cells and fibroblasts. Semi-quantitative analysis of IHC samples showed significantly increased expression of AGE (p < .0001) and RAGE (p = .02) in T2D samples compared with controls. The expression of NFKB (p < .0001), S100A12 (p < .0001) and COLIA1 (p = .01) genes were significantly higher in the T2D pulp, and multivariate logistic regression analysis showed that these findings were not affected by age. T2D may exert a similar glycation response in the dental pulp to other body sites. This could occur through activation of NF-κB pathways with a concomitant increase in genes associated with inflammation and collagen.

Carvacrol, sodyum arsenit kaynaklı ince bağırsak toksisitesini iyileştirmektedir

Arsenic is a very dangerous metal that is widely distributed in the environment as a result of anthropogenic and natural processes. On the other hand, it is thought that treatment with natural flavonoids may protect against arsenic toxicity. Carvacrol (CRV) is a naturally occurring phenolic compound with anti-inflammatory, anti-apoptotic, and antioxidant properties. This study aims to investigate the protective effects of CRV on sodium arsenite (SA)-induced intestinal toxicity. For this purpose, rats were randomly divided into five groups and administered SA and CRV orally for 14 days. The data indicate that when SA is administered, lipid peroxidation increase and antioxidant enzyme activities decrease. The administration of CRV ameliorated this impairment in the antioxidant defense system. Examining the expression levels of NF-kB and IL-1 revealed that inflammation increased with SA application but decreased with CRV administration. Moreover, the expression levels of Caspase-3 and Apaf1 increased in rats treated with SA, whereas the severity of apoptosis decreased when CRV was administered. In light of these findings, it is possible to state that CRV protects tissues from damage by presenting antioxidant, anti-inflammatory, and anti-apoptotic effects in SA-induced intestinal toxicity.

Effects and Mechanism of Epidural Administration of Triamcinolone Acetonide on Nerve Root Inflammation and Epidural Space Adhesion in Rabbits

This study aimed to investigate the effect and mechanism of triamcinolone acetonide on nerve root inflammation and epidural adhesion in rabbits. The experiment involved 30 male New Zealand white rabbits, which were divided into three groups: Model, Dexamethasone, and Triamcinolone. After establishing the rabbit model of nerve root inflammation and epidural adhesion, dexamethasone and triamcinolone acetonide were injected through the epidural catheter. The results showed that triamcinolone acetonide significantly reduced nerve root inflammation, epidural adhesion, and nerve root pain. It also decreased the mRNA expression levels of NF-KB, TLR9, IL-6, and TNF-α in spinal cord tissue and spinal nerve roots. Compared with the dexamethasone group, the triamcinolone group had a lower Rydell-Balaz score, indicating less epidural adhesion. The Paw Withdraw Threshold and Paw Withdraw Latency decreased significantly in the triamcinolone group compared to the model group, and there was no obvious nerve root inflammatory reaction in the triamcinolone group. Therefore, epidural triamcinolone acetonide injection may be an effective treatment to reduce nerve root inflammation, epidural adhesion, and nerve root pain by inhibiting the NF-KB/TLR9-mediated inflammatory response.

Curcumin-Loaded Nanocomposite Hydrogel Augments Functional Muscle Recovery and Shows Synergistic Effects with Post-Injury Treadmill Exercise in a Muscle Injury Model in Rats: An In Vitro and In Vivo Study

Herein, the synergistic healing activity of treadmill exercise and curcumin-delivering hydrogel was evaluated in a rat model of skeletal muscle injury. In Vitro experiments were utilized to characterize the filler material and the healing potential of the treatment strategy was investigated in a rat model of anterior tibialis muscle injury. Study showed excellent biocompatibility of the hydrogel system. In Vivo study showed that the developed hydrogel and treadmill sport regimen significantly augmented muscle function recovery as evidenced by histopathological analysis. Gene expression studies showed that tissue expression levels of NFK-B and collagen type 1 genes were significantly downregulated in the animals treated with curcumin-loaded hydrogels with and without exercise.

In vitro and in silico perspectives to explain anticancer activity of a novel syringic acid analog ((4-(1H-1, 3-benzodiazol-2-yl)-2, 6-dimethoxy phenol)) through apoptosis activation and NFkB inhibition in K562 leukemia cells

Syringic acid (SA) is an active carcinogenesis inhibitor; however, the low bioavailability and unstable functional groups hinder its activity. Here, a chemically synthesized novel SA analog (SA10) is evaluated for its anticancer activity using in-vitro and in-silico studies. K562 cell line study revealed that SA10 had shown a higher rate of inhibition (IC50 = 50.40 μg/mL) than its parental compound, SA (IC50 = 96.92 μg/mL), at 50 μM concentration. The inhibition ratio was also been evaluated by checking the expression level of NFkB and Bcl-2 and showing that SA10 has two-fold increase in the inhibitory mechanism than SA. This result demonstrates that SA10 acts as an NFkB inhibitor and an apoptosis inducer. Further, molecular docking and simulation have been performed to get insights into the possible inhibitory mechanism of SA and SA10 on NFkB at the atomistic level. The molecular docking results exemplify that both SA and SA10 bind to the active site of NFkB, thereby interfering with the association between DNA and NFkB. SA10 exhibits a more robust binding affinity than SA and is firmly docked well into the interior of the NFkB, as confirmed by MM-PBSA calculations. In a nutshell, the Benzimidazole scaffold containing SA10 has shown more NFkB inhibitory activity in K562 cells than SA, which could be helpful as an ideal therapeutic NFkB inhibitor for treating cancers.

Evaluation of the effects in the &lt;i&gt;in vitro&lt;/i&gt; system of synthetic thymic hexapeptide on the expression levels of NF-κB, IFNα/βR and CD119 neutrophilic granulocytes in patients with chronic herpes viral co-infections

Background. Strategies used by herpes viruses with human cells are complex and multifaceted. On one hand, inborn defects in antiviral immune defense have been unveiled, which also affect interferon (IFN) system underlying development of chronic recalcitrant relapsing viral infections such as remittent respiratory viral infections, herpesvirus infections, and papillomavirus infections. On the other hand, numerous viruses are able to damage both immune system and IFN network. During inborn and acquired defects in IFN network, inborn or induced mutation in gene products involved in signaling cascades aimed at upregulating gene expression responsible for IFN production are observed. One of the strategies used by diverse viruses is altering some signaling pathways resulting in activated transcription factors including nuclear factor NF-kB. However, antiviral mechanisms executed by neutrophilic granulocytes (NGs), particularly affecting NF-kB expression have not been elucidated. Aim of the study: to study in vitro features of NF-kB expression and number of neutrophilic granulocytes (NG) expressing membrane IFN/R and IFNR in patients with atypical chronic active herpes virus infections (AChA-HVI), followed by assessing an effect of arginyl-alpha-aspartyl-lysyl-valyl-tyrosyl-arginine hexapeptide (HP), a synthetic analogue of the active center of the thymopoietin (active substance of drug Imunofan, Russia), on the expression of NG NF-kB and IFN/R and IFNR. Materials and methods. We observed 25 patients of both sexes aged 23 to 64 years with AChA-HVI, manifested by chronic fatigue syndrome and cognitive disorders. Study design: stage 1 clinical, ELISA, PCR methods, FC was used. Stage 2 the in vitro experiment: 32 blood samples from 8 healthy adults and 375 blood samples from 25 patients with AChA-HVI were analyzed: % NG expressing NF-kB, IFN/R, IFNR and the relevant MFI levels by using FC before and after incubation with HP. Results. Our study demonstrated low level (MFI) of NF-kB expression in 100% NG associated with decreased % of NG expressing IFN/R and IFNR in all patients with AChA-HVI and low serum level for IFN and IFN in comparison with healthy individuals. In the in vitro experiment there was shown that 100% of NG expressed NF-kB after exposure to HP. However, only 48% patients (SG 2) restored NF-kB expression level (MFI) to normal range and 52% of cases (SG 1) had no response. HP increased % of NG expressing IFN/R in SG 2 and increased % of NG expressing IFNR in SG 1. Conclusions. It was shown, that influence of HP in vitro has ambiguous effects on the expression of NF-kB, % of NG expressing IFN/R and IFNR in patients with AChA-HVI. We assume that different NF-kB response to HP is associated with inborn or secondary NF-kB deficiency.

The ameliorating effect of Terminalia muelleri extract on oxidative stress–related factors in induced hepatocellular carcinoma rat model

The present study investigates the efficacy of Terminalia muelleri ethanol extract (TME) in alleviating the oxidative stress and inflammation that could be the main player of hepatocellular carcinoma (HCC) development and progression. Forty-eight male albino rats were divided into two main groups (32 rats for DENA administration; 16 rats for control groups); after the induction period of DENA administration (6 weeks), the HCC-induced rats were treated with TME, DOX, or their combination for eight weeks. The obtained results revealed that the DENA induced HCC resulted in remarkable dysregulation in the circulatory levels of TNF-α and IL-10 that accompanied by the initiation of free radicals with the generation of the oxidative stress, in addition to an increase in the mRNA expression levels of NFkB, MMP 9 and downregulation of mRNA expression level of TIMP1. The treatment of HCC induced rats with DOX, TME, or their combination alleviated and suppressed the progression of HCC in the following manner TME + DOX followed by TME then DOX. The obtained results revealed that TME was more valuable as an anti-cancer agent than DOX, employed in the current study as reference chemotherapy. T. muelleri extract attenuates inflammation, oxidative stress, and extracellular matrix proteolysis in hepatocellular carcinoma-induced rats.

Investigating the expression level of NF-KB and HIF1A genes among the inhabitants of two different background radiation areas in Ramsar, Iran

This study investigated the fluctuation of NF-KB and HIF-1a gene expression between inhabitants of a high-level background radiation area (HBRA) and a normal-level background radiation area (NBRA) of Ramsar, Iran. Sixty participants with the mean age of 48 ± 15 years were selected and divided into two groups. The group receiving a dose of ≤1.5 mGy/year (NBRA) was considered the control group and the target group (HBRA) received a dose of >1.5 mGy/year. These two groups were from neighbor regions to minimize socioeconomic differences between the participants. Blood samples were collected from each group and NF-KB and HIF-1a expression levels were compared using quantitative real-time PCR (qPCR) based on the stem loop method. The effects of residency duration in the respective areas and gender on the expression of NF-KB and HIF-1a was also examined. The HIF-1a expression level was statistically lower in the HLBRA region (P < 0.0002), while NF-KB expression was upregulated (P < 0.0001). Although the under-expression of HIF-1a in response to dose rate was significant in females (P < 0.0004), it was not different in males (P = 0.74), indicating a significant difference between sexes (P = 0.0047). The upregulation of NF-KB expression related to dose level was also significant for the female group (P < 0.0001), whereas it was not for the male group (P = 0.72). Notably and as expected, there was a significant relation between longer residency in the HBRA and HIF-1A under-expression (P < 0.026), while there was no effect of increasing residency time for NF-KB over-expression level (P = 0.29). The dwellers of the HBRA those noted that despite receiving an elevated radiation level were seemingly good in general health, showed some alterations in their molecular mechanisms, specifically HIF-1a and NF-KB expression levels. It is not clear if this is indicative of a beneficial adaptive response and more research is recommended.

MTOR/NF-κB signaling pathway protects hippocampal neurons from injury induced by intermittent hypoxia in rats

Objective To expound the roles of mTOR and NF-kB signaling pathway in intermittent hypoxia (IH)-induced damage of hippocampal neurons. Methods For rat experiments, mTOR inhibitor (Rapamycin, Rapa) and NF-κB signaling inhibitor (ammonium pyrrolidine dithiocarbamate, PDTC) were applied to inhibit mTOR and NF-κB signaling, respectively. For neuron experiments, hippocampal neurons from rat were successfully cultured. Different concentrations of Rapa and PDTC were added to the cultured hippocampal neurons. Rat or primary hippocampal neurons were exposed to normoxic or IH conditions after administration of Rapa and PDTC. The effects of Rapa and PDTC administration on learning and memory ability of rats and hippocampal injury after IH exposure were assayed by Morris water maze and H&E staining. Electron microscope was utilized to examine primary hippocampal neuron ultrastructure changes after IH exposure and Rapa or PDTC administration. The expressions of NF-κB-p65, IκBα, IKKβ, BDNF, TNF-α, IL-1β, PSD-95 and SYN in hippocampal neurons were examined. Results Compared with normal control rats or neurons, IH-treated group had elevated expression levels of NF-kB, TNF-α and IL-1β and suppressed expression level of BDNF, PSD-95 and SYN. These results were reversed upon pre-treatment with Rapa and PDTC. Furthermore, IκBα and IKKβ expressions were down-regulated in IH group. No notable difference was manifested in PDTC pre-treatment group, while a prominent increase was shown after Rapa pre-administration. Conclusion The administration of PDTC and Rapa could prevent IH-induced hippocampal neuron impairment, indicating that inhibition of the mTOR and NF-κB pathway may likely act as a therapeutic target for obstructive sleep apnea.

Metformin suppresses the proliferation and invasion through NF-kB and MMPs in MCF-7 cell line

Abstract Objective MCF-7 cells, a breast cancer cell line, are used for experiments of estrogen receptor (ER)-positive breast cancer and many sub-clones representing different classes of ER-positive tumors. We aimed to determine the efficacy of metformin, a potential anti-cancer agent, on the cell proliferation, and the expressions of NF-kB (p65), MMP-2 and MMP-9 in MCF-7 cell line. Materials and methods MCF-7 cells (human breast adenocarcinoma) were treated with elevating doses of metformin (0–50 mM) for 24 h. The anti-proliferative effect of metformin was studied by BrdU proliferation assay, and the expression levels of NF-kB (p65), MMP-2 and MMP-9 were analyzed by immunocytochemical staining. Results The percentage of cell proliferation was reduced significantly by 10 and 50 mM doses of metformin (p &lt; 0.001). The expression levels of nuclear NF-kB (p65), MMP-9 and MMP-2 were considerably reduced in 50 mM metformin treated cells while the expression of cytoplasmic NF-kB (p65) elevated compared to control group (p &lt; 0.05). Ten millimolar metformin also reduced expression of MMP-9 significantly (p &lt; 0.05). Conclusion Metformin may act on the proliferation, and the processes of invasion and metastasis of MCF-7 cells through blocking NF-kB, which is intensely expressed in breast cancer cells, and through diminishing the expression of MMP-2 and MMP-9 significantly.

Effects of crustacean hyperglycemic hormone (CHH) on regulation of hemocyte intracellular signaling pathways and phagocytosis in white shrimp Litopenaeus vannamei

Shrimps like other arthropods rely on innate immune system, and may have some form of adaptive immunity in defending against pathogens. Phagocytosis is one of the oldest cellular processes, serving as a development process, a feeding mechanism and especially as a key defense reaction in innate immunity of all multicellular organisms. It is confirmed that crustacean hyperglycemic hormone (CHH) is one of the most important neuropeptides produced by Neuro-endocrine Immune (NEI) regulatory network, which undertakes important roles in various biological processes, especially in immune function and stress response. In this study, the recombinant Litopenaeus vannamei CHH (rLvCHH) was obtained from a bacterial expression system and the intracellular signaling pathways involved in the mechanism of phagocytosis after rLvCHH injection was investigated. The results showed that the contents of adenylyl cyclase (AC), phospholipase C (PLC) and calmodulin (CaM) in hemocytes were increased significantly after rLvCHH injection. Furthermore, the mRNA expression levels of NF-kB family members (relish and dorsal) and phagocytosis-related proteins in hemocytes were basically overexpressed after rLvCHH stimulation, while the expression level of NF-kB repressing factor (NKRF) gene was down-regulated significantly. Eventually, the total hemocyte count and phagocytic activity of hemocyte were dramatically enhanced within 3 h. Collectively, these results indicate that shrimps L. vannamei could carry out a simple but ‘smart’ NEI regulation through the action of neuroendocrine factors, which could couple with their receptors and trigger the downstream signaling pathways during the phagocytic responses of hemocytes.

Rap2B promotes cell adhesion, proliferation, migration and invasion of human glioma.

Rap2B, a member of the GTP-binding proteins, is generally up-regulated in numerous types of tumors. Nevertheless, the influence and regulatory mechanisms of Rap2B in gliomas are still not corroborated. Therefore, we analyzed the expression of Rap2B in glioma tissues and cells, and researched its significance in adhesion, proliferation, migration and invasion of the glioma cell line. We analyzed the expression of Rap2B in different pathologic grades of glioma tissues by tissue microarray and immunohistochemistry. We assessed the expression of Rap2B in glioma tissue and non-tumor tissue by Western blot. And the expression of Rap2b protein in glioma cells and normal human astrocytes (NHA) was detected by Western blot. In addition, we disclosed the effect of Rap2B knockdown on cell adhesion, proliferation, migration and invasion by using cell attachment assay, CCK-8 assay, cell migration assay and Wound Healing assay, cell invasion assay, respectively. Western blot was used to detect the changes of expression level of NF-kB, MMP-2 and MMP-9 protein when downregulated the expression of Rap2B. The tissue microarray immunohistochemical results of glioma showed that the expression of Rap2B had no significant correlations between Rap2B expression and the clinicopathologic variables, including patient age (P=0.352), gender (P=0.858), WHO Grade (P=0.693) and histology type (P=0.877). Western blot analysis showed that the glioma tissue had a dramatically increase of Rap2B expression compared with the non-tumor tissues (P<0.01). And the expression of Rap2B was markedly up-regulated in all 5 glioma cell lines compared with that in normal human astrocytes (NHA) (P<0.01). We found that the ability of adhesion, proliferation, migration and invasion of glioma cells were significantly decreased after downregulated Rap2B expression compared with the control group (P<0.05). In addition, Western blot results showed that the expression levels of NF-kB, MMP-2 and MMP-9 in the interference group were significantly lower than those in the negative control group (P<0.05). Rap2B expression is up-regulated in glioma tissues and glioma cell lines. Knockdown of Rap2B inhibits glioma cells' adhesion and proliferation in vitro. Knockdown of Rap2B inhibits glioma cells' migration in vitro. Knockdown of Rap2B inhibits glioma cells' invasion and MMPs activity through NF-kB pathway.

Lipoxin A4 Methyl Ester Reduces Early Brain Injury by Inhibition of the Nuclear Factor Kappa B (NF-κB)-Dependent Matrix Metallopeptidase 9 (MMP-9) Pathway in a Rat Model of Intracerebral Hemorrhage.

BackgroundIntracerebral hemorrhage (ICH) is associated with inflammation and disruption of the blood-brain barrier (BBB). Lipoxin A4 methyl ester (LXA4 ME), is a stable synthetic analog of lipoxin with anti-inflammatory properties. This study aimed to investigate the effects of LXA4 ME in a rat model of ICH.Material/MethodsMale Sprague-Dawley rats (n=120), between 12–13 weeks of age, were divided into the sham group (sham-operated), the vehicle-treated group (ICH+vehicle), the LXA4 ME-L group (ICH+low-dose LXA4 ME, 10 ng/d), and the LXA4 ME-H group (ICH+high-dose LXA4 ME, 100 ng/d). The ICH model was created by injection of autologous blood into the right basal ganglia. LXA4 ME was injected into the ventricle 10 min after the development of ICH. A modified neurological severity score (mNSS), rotarod latencies, and brain water content were used to evaluate the rats. The TUNEL assay measured neuronal cell death. Western blot was used to measure protein expression of nuclear factor kappa B (NF-κB), matrix metalloproteinase-9 (MMP-9), zonula occludens-1 (ZO-1), and claudin-5.ResultsIn the rat model of ICH, treatment with LXA4 ME reduced the levels of proinflammatory cytokines, improved neurologic function, reduced neuronal apoptosis, and reduced cerebral edema associated with damage to the BBB, and reduced the expression levels of NF-κB, MMP-9, ZO-1, and claudin-5.ConclusionsIn a rat model of ICH, treatment with LXA4 reduced early brain injury and protected the BBB by inhibiting the NF-κB-dependent MMP-9 pathway.

Protective effects of dioscin against fructose-induced renal damage via adjusting Sirt3-mediated oxidative stress, fibrosis, lipid metabolism and inflammation

In the present work, the effects and possible mechanisms of dioscin, one natural product from the famous vegetable Dioscoreae rhizoma (Shanyao in Chinese), against high fructose-induced renal injury in rats were tested. The results showed that dioscin significantly restored fructose-induced renal injury by decreasing the levels of Cr, BUN, and rehabilitating histopathological changes. In addition, dioscin markedly adjusted the levels of MDA, SOD and GSH-Px, reduced ROS level in renal tissue, and decreased the levels of TG, FFA, α-SMA and COL1A. Mechanistic study showed that dioscin significantly up-regulated the expression levels of Sirt3, SOD2, and then suppressed inflammation by decreasing the expression levels of NF-kB, HMGB1, c-Jun, c-Fos, COX2, TNF-α, IL-1β and IL-6. Furthermore, dioscin-caused high levels of Sirt3 and SOD2 attenuated oxidative stress by regulating the expression levels of Nrf2, GST, Keap1, regulated lipid metabolism by controlling the expression levels of SREBP-1c, SCD-1, FASn, ACC, CPT1, and adjusted TGF-β1/Smad signal to inhibit renal fibrosis. In summary, dioscin showed protective effects against fructose-induced renal damage via adjusting Sirt3-mediated oxidative stress, renal fibrosis, lipid metabolism and inflammation, which should be considered as one candidate to treat renal injury in the future. We also suggest that the patients with renal injury can take more Shanyao for the therapy and treatment.

Evaluation of hydro-alcoholic extract of Eclipta alba for its anticancer potential: An in vitro study

Eclipta alba is traditionally used as hepatoprotective agent. The study was designed to explore its antiproliferative activity on liver and other related cancer. The present study was designed to assess and establish the role of Eclipta alba as anti-cancer agent using HepG2, C6 glioma and A498 cell lines as model system. Antiproliferative and cytotoxic effects of the Eclipta alba hydroalcoholic extract (EAE) was determined using MTT assay. The expression level of NF-kB was analysed by western blotting and RT PCR. Gelatin zymography was done for gelatinase matrix metalloproteinases (MMP-2 and 9) analysis. EAE inhibited the cell proliferation in dose dependent manner in HepG2, A498 and C6 glioma cell lines with an IC50 of 22±2.9, 25±3.6 and 50±8.7 μg/ml, respectively. The expression of MMP (2 and 9) was down-regulated with EAE treatment. DNA damage was observed following 72h of extract treatment, leading to apoptosis. Additionally, the expression level of NF-kB was evaluated with western blotting and RT-PCR and was found to be down-regulated/inactivated. The data establish the existence of anti-proliferative, DNA damaging and anti-metastasis properties in EAE which is yet unexplored and hold high therapeutic impact.

Peroxiredoxin 2 Inhibits Granulosa Cell Apoptosis During Follicle Atresia Through the NFKB Pathway in Mice1

Peroxiredoxin 2 (PRDX2) has been known to act as an antioxidant enzyme whose main function is H(2)O(2) reduction in cells. We aimed to study the expression patterns of PRDX2 in mouse ovaries and explore the function of this protein in apoptosis of granulosa cells (GCs). We found that the expression of the PRDX2 protein in atretic follicle GCs was markedly higher than in healthy follicle GCs. In vitro, the transfection of siRNA targeting the Prdx2 gene inhibited the proliferation and induced the apoptosis of primary cultured GCs. Furthermore, suppression of PRDX2 resulted in the augmentation of endogenous H(2)O(2), and the ability to eliminate the exogenous H(2)O(2) was attenuated. The expression of PRDX2 and nuclear factor kappa-light-chain-enhancer of activated B cells (NFKB), whose activity was inhibited by binding to IKB, increased in GCs treated with various concentrations of H(2)O(2) for 30 min. However, no significant change in cytoplasmic IKB expression was observed. At 2 h after treatment with H(2)O(2), nuclear NFKB expression level was reduced, cytoplasmic IKB expression was increased, and PRDX2 expression was unchanged. Silencing of the Prdx2 gene caused early changes in NFKB and IKB expression in the primary cultured GCs compared to that in control cells. Taken together, these data suggest that PRDX2 plays an important role in inhibiting apoptosis in GCs and that PRDX2 actions may be related to the expression of NFKB and IKB.