mRNA Expression Levels Of TNF-α Research Articles

Effects of Continuous LPS Induction on Oxidative Stress and Liver Injury in Weaned Piglets

Due to imperfections in their immune and digestive systems, weaned piglets are susceptible to invasions of the external environment and diseases, especially bacterial infections, which lead to slow growth, tissue damage, and even the death of piglets. Here, a model of weaned piglets induced by Escherichia coli lipopolysaccharide (LPS) was established to explore the effects of continuous low-dose LPS induction on the mechanism of liver injury. A total of forty-eight healthy 28-day-old weaned piglets (weight = 6.65 ± 1.19 kg) were randomly divided into two groups: the CON group and LPS group. During the experimental period of thirteen days, the LPS group was injected intraperitoneally with LPS (100 μg/kg) once per day, and the CON group was treated with the same volume of 0.9% NaCl solution. On the 1st, 5th, 9th, and 13th days, the serum and liver of the piglets were collected for the determination of serum biochemical indexes, an antioxidant capacity evaluation, and histopathological examinations. In addition, the mRNA expression levels of the TLR4 pathway and inflammatory cytokines were detected. The results showed that the activities of aspartate aminotransferase (AST), alanine aminotransferase (ALT), and alkaline phosphatase (ALP) in the serum increased after LPS induction. The activities of total antioxidant capacity (T-AOC) and glutathione peroxidase (GSH-Px) in the serum and liver homogenate of the LPS group were lower than those of the CON group, while the malondialdehyde (MDA) content in the serum and the activities of catalase (CAT) and superoxide dismutase (SOD) in the liver of the LPS group were higher than those in the CON group. At the same time, morphological impairment of the livers occurred, including hepatocyte caryolysis, hepatocyte vacuolization, karyopycnosis, and inflammatory cell infiltration, and the mRNA expression levels of TLR4, MyD88, NF-κB, TNF-α, IL-6, and IL-10 were upregulated in the livers after LPS induction. The above results were more obvious on the 1st and 5th days of LPS induction, while the trend during the later period was not significant. It was concluded that the oxidative stress and liver injury occurred at the early stage of LPS induction, while the liver damage weakened at the later stage. The weaned piglets probably gradually developed tolerance to the endotoxin after the continuous low-dose induction of LPS.

Isolation, Characterization and Immunomodulatory Activity Evaluation of Chrysolaminarin from the Filamentous Microalga Tribonema aequale.

The aim of this study is to investigate the differences in the accumulation capacity of chrysolaminarin among six Tribonema species and to isolate this polysaccharide for immunomodulatory activity evaluation. The results showed that T. aequale was the most productive strain with the highest content and productivity of chrysolaminarin, which were 17.20% (% of dry weight) and 50.91 mg/L/d, respectively. Chrysolaminarin was then extracted and isolated from this alga, and its monosaccharide composition was mainly composed of a glucose (61.39%), linked by β-D-(1→3) (main chain) and β-D-(1→6) (branch chain) glycosidic bonds, with a molecular weight of less than 6 kDa. In vitro immunomodulatory assays showed that it could activate RAW264.7 cells at a certain concentration (1000 μg/mL), as evidenced by the increased phagocytic activity and upregulated mRNA expression levels of IL-1β, IL6, TNF-α and Nos2. Moreover, Western blot revealed that this polysaccharide stimulated the phosphorylation of p-65, p-38 and JNK in NF-κB and MAPK signaling pathways. Overall, these findings provide a reference for the further development and utilization of algae-based chrysolaminarin, while also offering an in-depth understanding of the immunoregulatory mechanism.

Sophora flavescens Aiton methanol extract exerts anti-inflammatory effects via reduction of Src kinase phosphorylation

Ethnopharmacological relevanceSophora flavescens Aiton (Family: Leguminosae), an herbal plant, has been used in East Asian home remedies for centuries for treating ulcers, skin burns, fevers, and inflammatory disorders. In addition, the dried root of S. flavescens was also applied for antipyretic, analgesic, antihelmintic, and stomachic uses. Aim of studyNonetheless, how this plant can show various pharmacological activities including anti-inflammatory responses was not fully elucidated. In this study, therefore, we aimed to investigate the curative effects of S. flavescens on inflammation and its molecular mechanism. Materials and methodsFor reaching this aim, various in vitro and in vivo experimental models with LPS-treated RAW264.7 cells, HCl/EtOH-induced gastric ulcer, and LPS-triggered lung injury conditions were employed and anti-inflammatory activity of S. flavescens methanol extract (Sf-ME) was also tested. Fingerprinting profile of Sf-ME was identified via LC-MS analysis. Its anti-inflammatory molecular mechanism was also examined by immunoblotting analysis. ResultsNitric oxide production and mRNA expression levels of iNOS, COX-2, IL-1β, and TNF-α were decreased. Additionally, phosphorylation of Src in the signaling cascade was decreased, and activities of the transcriptional factor NF-κB were reduced as determined by a luciferase reporter assay. Moreover, in vivo, gastritis and lung injury lesions were attenuated by Sf-ME. ConclusionTaken together, these findings suggest that Sf-ME could be a potential anti-inflammatory therapeutic agent via suppression of Src kinase activity and regulation of IL-1β secretion.

Correlations of Myeloperoxidase (MPO), Adenosine deaminase (ADA), C–C motif chemokine 22 (CCL22), Tumour necrosis factor alpha (TNFα) and Interleukin-6 (IL-6) mRNA expression in the nasopharyngeal specimens with the diagnosis and severity of SARS-CoV-2 infections

ABSTRACT Cytokine dynamics in patients with coronavirus disease 2019 (COVID-19) have been studied in blood but seldomly in respiratory specimens. We studied different cell markers and cytokines in fresh nasopharyngeal swab specimens for the diagnosis and for stratifying the severity of COVID-19. This was a retrospective case-control study comparing Myeloperoxidase (MPO), Adenosine deaminase (ADA), C–C motif chemokine ligand 22 (CCL22), Tumour necrosis factor alpha (TNFα) and Interleukin-6 (IL-6) mRNA expression in 490 (327 patients and 163 control) nasopharyngeal specimens from 317 (154 COVID-19 and 163 control) hospitalized patients. Of the 154 COVID-19 cases, 46 died. Both total and normalized MPO, ADA, CCL22, TNFα, and IL-6 mRNA expression levels were significantly higher in the nasopharyngeal specimens of infected patients when compared with controls, with ADA showing better performance (OR 5.703, 95% CI 3.424–9.500, p < 0.001). Receiver operating characteristics (ROC) curve showed that the cut-off value of normalized ADA mRNA level at 2.37 × 10–3 had a sensitivity of 81.8% and specificity of 83.4%. While patients with severe COVID-19 had more respiratory symptoms, and elevated lactate dehydrogenase, multivariate analysis showed that severe COVID-19 patients had lower CCL22 mRNA (OR 0.211, 95% CI 0.060–0.746, p = 0.016) in nasopharyngeal specimens, while lymphocyte count, C-reactive protein, and viral load in nasopharyngeal specimens did not correlate with disease severity. In summary, ADA appears to be a better biomarker to differentiate between infected and uninfected patients, while CCL22 has the potential in stratifying the severity of COVID-19.

TNF-α and IL-1β Promote Renal Podocyte Injury in T2DM Rats by Decreasing Glomerular VEGF/eNOS Expression Levels and Altering Hemodynamic Parameters.

Diabetic nephropathy (DN) is a serious microvascular complication in those with type 2 diabetes mellitus (T2DM). Evidence confirms that serum tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) in the T2DM stage are proposed as prognostic markers for DN development, but it is unclear how they affect renal podocyte-associated nephrin and WT-1 expression. In the presence of podocyte injury, glomerular vascular endothelial growth factor (VEGF), endothelial nitric oxide synthase (eNOS) and hemodynamic parameters are dysregulated. The current research aimed to clarify the relationship of TNF-α and IL-1β with podocyte injury by altering VEGF/eNOS expression and hemodynamic parameters. A high-fat diet/streptozotocin-induced DN rat model was established. Serum TNF-α and IL-1β levels were tracked in the pre-T2DM, T2DM and DN stages. In the DN stage, the mRNA and protein expression levels of renal TNF-α, IL-1β, VEGF, eNOS, nephrin and WT-1 were studied. Renal hemodynamic parameters, including peak systolic velocity, end-diastolic flow velocity and mean velocity were measured with a color Doppler ultrasound technique. Compared to those in the normal control (CTL) group, serum TNF-α and IL-1β levels increased significantly in the pre-T2DM stage (obesity, insulin resistance and hyperlipidemia), T2DM stage (hyperglycemia) and DN stage (abnormal renal functions) (all: P < 0.05) in the DN group. Serum TNF-α and IL-1β levels in the T2DM stage were significantly higher than those in the pre-T2DM stage (two: P < 0.05). Compared to the CTL group, renal nephrin, WT-1, TNF-α, IL-1β, eNOS and VEGF expression and hemodynamic parameters in the DN stage all showed significant differences separately (all: P < 0.05). Increased serum and renal TNF-α and IL-1β levels played important roles in reducing renal nephrin and WT-1 expression levels, which may be related to the fact that the former affected renal VEGF/eNOS expression and blood flow parameters in the DN rats.

Globular adiponectin-mediated vascular remodeling by affecting the secretion of adventitial-derived tumor necrosis factor-α induced by urotensin II.

In this study, we explored how adiponectin mediated urotensin II (UII)‍-induced tumor necrosis factor-‍α (TNF-‍α) and α‍-smooth muscle actin (α‍-SMA) expression and ensuing intracellular signaling pathways in adventitial fibroblasts (AFs). Growth-arrested AFs and rat tunica adventitia of vessels were incubated with UII and inhibitors of signal transduction pathways for 1‍‒‍24 h. The cells were then harvested for TNF-α receptor (TNF-‍α-R) messenger RNA (mRNA) and TNF-‍α protein expression determination by reverse transcription-polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA), respectively. Adiponectin and adiponectin receptor (adipoR) expression was measured by RT-PCR, quantitative real-time PCR (qPCR), immunohistochemical analysis, and cell counting kit-8 (CCK-8) cell proliferation experiments. We then quantified TNF-α and α-SMA mRNA and protein expression levels by qPCR and immunofluorescence (IF) staining. RNA interference (RNAi) was used to explore the function of the adipoR genes. To investigate the signaling pathway, we applied western blotting (WB) to examine phosphorylation of adenosine 5'-monophosphate (AMP)‍-activated protein kinase (AMPK). In vivo, an adiponectin (APN)‍-knockout (APN-KO) mouse model mimicking adventitial inflammation was generated to measure TNF-α and α‍-SMA expression by application of qPCR and IF, with the goal of gaining a comprehensive atlas of adiponectin in vascular remodeling. In both cells and tissues, UII promoted TNF-α protein and TNF-α-R secretion in a dose- and time-dependent manner via Rho/protein kinase C (PKC) pathway. We detected marked expression of adipoR1, T-cadherin, and calreticulin as well as a moderate presence of adipoR2 in AFs, while no adiponectin was observed. Globular adiponectin (gAd) fostered the growth of AFs, and acted in concert with UII to induce α-SMA and TNF-α through the adipoR1/T-cadherin/calreticulin/AMPK pathway. In AFs, gAd and UII synergistically induced AMPK phosphorylation. In the adventitial inflammation model, APN deficiency up-regulated the expression of α-SMA, UII receptor (UT), and UII while inhibiting TNF-‍α expression. From the results of our study, we can speculate that UII induces TNF‍-‍α protein and TNF-‍α‍-R secretion in AFs and rat tunica adventitia of vessels via the Rho and PKC signal transduction pathways. Thus, it is plausible that adiponectin is a major player in adventitial progression and could serve as a novel therapeutic target for cardiovascular disease administration.

Impact and mechanism of CHL1 in insulin resistant adipocytes and insulin resistant mouse model induced by high glucose and high fat

Objective: To investigate the role and mechanism of cell adhesion molecule L1 like (CHL1) in insulin resistant adipocytes and insulin resistant mouse model induced by high glucose and high fat. Methods: The 3T3-L1 preadipocytes were randomly divided into control group (transfected with empty vector) and CHL1 overexpression group (transfected with CHL1 vector), cells were then induced to mature adipocytes by insulin, and insulin resistance was then induced by high sugar and high fat. The glucose content was measured to determine the glucose consumption of cells from the two groups. Protein expression levels of CHL1 and glucose transporter 4 (GLUT4), serine/threonine protein kinase (AKT) phosphorylation levels were detected by Western blot (WB), the mRNA expression levels of TNF-α and IL-6 were detected by real-time quantitative PCR (RT-qPCR). 24 C57BL/6 adult male mouse were randomly divided into conventional diet group (regular group), high-fat diet group (high-fat group), empty vector overexpression+high-fat group and CHL1 overexpression+high-fat group (n=6 each group). CHL1 overexpression was induced by tail vein injection of lentivirus. Four months later, mice were sacrificed, body weight was determined, and the epididymal white adipose tissue was collect. Hematoxylin-eosin staining (HE) was used to observe the pathology of mouse epididymal white adipose tissue, the expression of CHL1 was evaluated by immunohistochemical staining(IHC), RT-qPCR was used to detect the mRNA expression levels of CHL1, TNF-α and IL-6 in mouse epididymal white adipose tissue. Results: In vitro, glucose consumption was significantly higher in the CHL1 overexpression group than in the control group (P<0.05), and the protein expressions of CHL1 and GLUT4 were higher in the CHL1 overexpression group than those in the control group (P<0.01), and the mRNA expressions levels of TNF-α and IL-6 were lower in the CHL1 overexpression group than those in the control group (P<0.01). In vivo, the body weight and epididymal white adipose tissue of mouse were higher in the high-fat group and the empty vector overexpression+high-fat group than those in the conventional group (P<0.01), which were lower in the CHL1 overexpression+high fat group than in the empty vector overexpression+high fat group (P<0.01). HE results showed that the volume of epididymal white adipocytes was larger in the high-fat group and the overexpression control+high-fat group than that in the conventional group, which was smaller in the CHL1 overexpression+high fat group than in the empty vector overexpression+high fat group (P<0.01). The mRNA expression levels of IL-6 and TNF-α in epididymal white adipose tissue of mice were higher in the high-fat group and the empty vector overexpression+high-fat group than those in the conventional group (P<0.01), which were lower in the CHL1 overexpression+high fat group than in the empty vector overexpression+high fat group (P<0.05). IHC results showed that protein expression of CHL1 in epididymal white adipose tissue was lower in the high-fat group and the empty vector overexpression+high-fat group than in regular group, which was upregulated in the CHL1 overexpression+high fat group than in the empty vector overexpression+high-fat group (P<0.01). RT-qPCR results showed that mRNA expression of CHL1 in epididymal white adipose tissue was lower in the high-fat group and the empty vector overexpression+high-fat group than in regular group (P<0.01), which was higher in the CHL1 overexpression+high fat group than in the empty vector overexpression+high fat group (P<0.01). Conclusion: Overexpression of CHL1 can improve insulin resistance in adipocytes and mouse insulin resistance model induced by high glucose and high fat, and the beneficial effects might be mediated by the inhibition of AKT activation and the reduction of related inflammatory responses.

Protective Role of the Toll-Like Receptor 5 Agonist KMRC011 against Murine Colitis Induced by Citrobacter rodentium and Dextran Sulfate Sodium.

This study aimed to identify the therapeutic ability of a novel toll-like receptor (TLR) 5 agonist, KMRC011, on ulcerative colitis induced by Citrobacter rodentium and dextran sulfate sodium in a C57BL/6N mouse model. Ulcerative colitis was induced in the mice by the oral administration of 1% dextran sulfate sodium in sterile drinking water for seven days ad libitum, followed by C. rodentium infection on the seventh day by intra-gastric administration (DSS-CT group). KMRC011 was administered intramuscularly at both 24 h and 15 min before (Treatment 1 group), and at both 15 min and 24 h after (Treatment 2 group) the C. rodentium infection. The length of the large intestine and histopathological counts were significantly greater and mucosal thickness was significantly thinner in the Treatment 1 group compared to the DSS-CT and Treatment 2 groups. Il-6 and Il-10 mRNA expression levels were upregulated, while Ifn-γ and Tnf-α mRNA expression levels were significantly downregulated in the Treatment 1 group, compared to the DSS-CT group. NF-κB p65 expression level was elevated due to ulcerative colitis in the DSS-CT group, but was significantly downregulated in the Treatment 1 group. Overall, KMRC011 showed protective effects against murine colitis by inhibiting NF-κB signaling.

Neuroprotective effects of etanercept on diabetic retinopathy via regulation of the TNF-α/NF-κB signaling pathway

Purpose: To study the influence of etanercept on diabetic retinopathy in rats via tumor necrosis factor alpha (TNF-α)/nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) signaling pathway. &#x0D; Methods: Thirty-six Sprague-Dawley (SD) rats were randomly divided into normal, model and etanercept groups. The expression of Caspase-3 was determined by immunohistochemistry, while the relative protein and mRNA expression levels of TNF-α and NF-κB were determined by Western blotting and quantitative polymerase chain reaction, respectively. Besides, the contents of TNF-α and interleukin-1 beta (IL-1β) were evaluated using enzyme-linked immunosorbent assay (ELISA), while cell apoptosis was assessed by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL). &#x0D; Results: Immunohistochemical studies showed that the mean optical density of tissues positive for caspase-3 in both the model and etanercept groups were significantly higher than in the normal group (p &lt; 0.05), while the mean optical density in the etanercept group was significantly lower than that in the model group (p &lt; 0.05). The protein expression levels of TNF-α and NF-κB in the etanercept group were significantly lower than those in the model group (p &lt; 0.05). Furthermore, mRNA expressions of TNF-α and NF-κB declined in the etanercept group (p &lt; 0.05); in addition, TNF-α, and IL-1β levels in the etanercept group were lower than in the model group (p &lt; 0.05). Cell apoptosis in the etanercept group was also lower than in the model group. &#x0D; Conclusion: Etanercept suppresses TNF-α/NF-κB signaling pathway thereby repressing inflammation and cell apoptosis in diabetic retinopathy rats. Therefore, etenercept’s neuroprotective effect may potentially be useful in developing a suitable therapy for diabetic neuropathy.

Beneficial Effect of Paeonol on Antibiotic-Associated Inflammatory Response in Mice with Diarrhea.

Diarrhea is a common adverse effect of antibiotics particularly that acts on anaerobes. Moutan Radicis Cortex (MRC) is an herbal medicine used for its anti-inflammatory and antibacterial actions. The purpose of this study was to analyze the active components of MRC to determine their effect on antibiotic-associated diarrhea (AAD) and anti-inflammatory effects. Of the various components of MRC, seven compounds (gallic acid, oxypaeoniflorin, paeoniflorin, ethyl gallate, benzoic acid, benzoylpaeoniflorin, paeonol) were identified and assessed for anti-inflammation effects. Paeonol was found to effectively reduce nitric oxide production and levels of IL-6 and TNF-α in a concentration-dependent manner. Paeonol also effectively reduced the mRNA expression level of IL-6, IL-1β, and TNF-α. Western blotting analysis confirmed the reduction of COX-2 and NF-κB levels; p-p38 MAPK levels increased in the presence of a low concentration (25 μM) of paeonol but decreased in the presence of a high concentration (50 μM). In the mouse model of lincomycin-induced AAD, all experimental groups treated with paeonol (25, 50, and 100 mg/kg concentrations) showed diminished diarrhea status scores. Finally, the expression levels of TNF-α and IL-4 were reduced compared with those in the control group. Therefore, paeonol may have active compounds of MRC to alleviate the diarrhea symptoms of AAD and reduce inflammatory mediators. Other components of the MRC extract could contribute to its known anti-inflammatory and antibacterial activity and should be tested for their possible activity.

Therapeutic Effect of Artesunate on Influenza A Viral Pneumonia

To investigate the therapeutic effect of artesunate (ART) on influenza A viral pneumonia. A total of 36 mice were evenly and randomly assigned to six groups, a normal control group (C group), a solvent control group (M group, 10% DMSO), a positive drug group (P group, oseltamivir, 1.25 mg/kg/day), ART high-dose group (ART-G group, 120 mg/kg/day), ART medium-dose group (ART-Z group, 60 mg/kg/day), and ART low-dose group (ART-D group, 30 mg/kg/day). Except for group C, which did not receive any influenza A virus intervention or intraperitoneal injection, mice in the five other groups were infected with influenza A virus through intranasal drip. Then, after 12 hours, mice in the five other groups received intraperitoneal injection of the assigned drugs and dosage once a day. The signs, body weight, and survival of the mice were observed over the course of treatment. After 7 days of treatment, the lung tissue of the mice was collected and weighed, and the lung index was calculated accordingly. HE staining was performed to observe the pathological changes in the lung tissue. The mRNA and protein expression levels of Toll-like receptor 4 (TLR4), nuclear factor kappa-B (NF-κB [p65]), tumor necrosis factor α (TNF-α), interleukin-6 (IL-6), and IL-1β were examined with RT-qPCR and Western blot, respectively. Compared with those in C group, mice in the M group had worse physical signs and lower body mass and survival, increased lung index, severe pathological changes in lung tissue, and increased levels of TLR4, NF-κB (p65), TNF-α, IL-6 and IL-1β mRNA and protein expression in their lung tissue ( P<0.05). Compared with those in M group, the mice in the ART groups had better physical signs, higher body mass and survival rate, decreased lung index, improvement of pathological changes in the lung tissue, and decreased levels of level of TLR4, NF-κB (p65), TNF-α, IL-6 and IL-1β mRNA and protein expression in the lung tissue ( P<0.05). Furthermore, the most prominent changes in these indexes were observed in the ART-G group. ART has therapeutic effects on influenza A viral pneumonia, and the mechanisms are related to the inhibition of TLR4/p65 signaling pathway activation and anti-inflammation.

Long-term low-dose microcystin-LR exposure induces renal injury in mice by activating PI3K/AKT signaling pathway

To investigate the toxic effect of long-term low-dose exposure to microcystin-LR (MC-LR) on kidney and its underlying molecular mechanism. Forty male C57BL/6 mice were randomized into 4 groups for exposure to 0, 1, 60, and 120 μg/L MC-LR (mixed in drinking water) for 12 months, and the body and kidney weight changes and renal pathologies of the mice were observed. The renal function indexes, the mRNA expression levels of IL-6, TNF-α and IL-10, and relative expression levels of PI3K/AKT pathway proteins in the kidney of the mice were detected. These parameters were also detected in HEK293 cells treated with MC- LR, LY294002, or both. The overall trend of body weight changes was consistent among the 4 groups of mice, and their kidney mass and kidney index underwent no significant changes. In mice exposed to 60 and 120 μg/L MC-LR, obvious renal structural damage and significant elevation of the BUN and SCr levels were observed (P < 0.05) with up-regulated levels of IL-6 and TNF-α mRNA and increased protein expressions of p-PI3K/PI3K and p-AKT/AKT in the renal tissues (P < 0.05). IL-10 mRNA expression was significantly decreased in all the exposure groups (P < 0.05). The levels of BUN and Cr increased significantly in MC-LR-treated HEK293 cells and decreased in cells treated with both MC-LR and LY294002 (P < 0.05). The mRNA expression levels of IL-6 and TNF-α increased and the level of IL-10 mRNA decreased obviously in MC-LR-treated cells, and the opposite changes were observed in the cells with the combined treatment (P < 0.05). The proteins levels of p-PI3K/PI3K and p-AKT/AKT were significantly up-regulated in MC-LR group and down-regulated in the combined treatment group (P < 0.05). MC- LR can activate inflammatory response and induce renal structural and functional damages in mice by activating the PI3K/AKT signaling pathway.

Protective Activities of Scutellarin against Alcohol-Induced Acute Kidney Injury.

Acute alcohol consumption has adverse effects in the kidney, resulting in kidney damage and disease, which are typically accompanied by oxidation and inflammation. Scutellarin (SCU) is the major effective ingredient of breviscapine and its anti-inflammation and antioxidant efficacy has been previously reported. The present study revealed the protective effective of SCU as therapeutic medicine against alcohol-induced inflammation and oxidative stress, leading to acute kidney injury (AKI). The AKI model was established by giving 50 % ethanol (12 mL/kg) via lavage. Kidney tissues were collected and used for histopathology analysis, biochemical assays and qRT-PCR analysis. The therapeutic effects of SCU were evaluated by observing pathological changes from HE-stained kidney tissues. Additionally, the anti-inflammation activity of SCU was evaluated by measuring the relative mRNA expression levels of Tnf-α, Il-1β, Il-6 and the activity of iNOS. The antioxidant capacity was assessed by measuring the lipid peroxidation marker 'MDA' and antioxidant enzymes activity of SOD, CAT and GSH-Px. The results showed that serious swelling and damage occurred in the renal tubular epithelium of alcohol intake group, accompanying with glomerular atrophy, necrosis and increase of inflammatory infiltration. SCU treatment significantly reduced the damage of diseased renal tubular epithelium and glomerular, and less inflammatory cell emerged. The inflammation cytokines expression levels were elevated and oxidative stress index decreased after alcohol intake compared to the control group. In conclusion, inflammation and oxidative stress occur in the kidney after acute and excessive alcohol intake, SCU exhibited protective roles via its anti-inflammation and antioxidant activity in AKI.

The effect of hydro alcoholic extract of Nigella sativa seeds on inflammatory mediators in C6 glioma cell line

Nigella sativa (N. sativa) is a highly valued nutritional plant, which has long been used in traditional medicine to treat a variety of human diseases. The multifaceted pharmacological impacts of N. sativa, such as attenuating oxidative stress and inflammation, make it a suitable therapeutic candidate against cardiovascular, hepatic, and neurological disorders as well as cancer. Therefore, the current study aimed to evaluate the effect of the hydroalcoholic extract of N.sativa seeds on several pro-inflammatory cytokines in the C6 glioma cell line and to compare it with the effect of the extract on the normal fibroblast cell line. C6 and fibroblast cell lines were treated with the extract of N.sativa seeds, and 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay was performed to determine the half-maximal inhibitory concentration (IC50) after 72h of treatment. Real-time polymerase chain reaction (RT-PCR) was carried out to assess the expression levels of interleukin (IL)-6, IL-10, tumor necrosis factor-alpha (TNF-α), and transforming growth factor- β1 (TGF-β1) at the mRNA level in both cell lines after 72h of treatment with non-toxic and IC50 concentrations obtained from C6 cell line. The IC50 values for the hydroalcoholic extract of N.sativa seeds were 260±20μg/mL in the C6 cell line and 398±27μg/mL in fibroblast cells. The real-time PCR results indicated that the treatment of C6 and fibroblast cells with the extract at the IC50 value of N.sativa in C6 for 72h could increase the mRNA expression levels of IL-10 and reduce the mRNA expression levels of IL-6, TNF-α, and TGF-β1 in C6 and fibroblast cells. The N.sativa extract showed a higher anti-inflammatory effect on C6 cells in comparison with fibroblast cells. Regarding the anti-inflammatory effect of Nigella sativa in C6 cell line, it may be considered a promising candidate to fortify antitumor actions in combination with other therapeutic options in the treatment of patients with GBM.

Anti-inflammatory potential of delta-9-tetrahydrocannabinol in hyperinsulinemia: an experimental study.

Hyperinsulinemia (HI) means that the amount of insulin in the blood is higher than normal and is often associated with type 2 diabetes. It is known that delta-9-tetrahydrocannabinol (THC) obtained from a medicinal plant, Cannabis sativa, has therapeutic effects on many diseases. This study aimed to investigate the effects of THC on inflammatory and oxidant status in rat pancreas with HI. Rats were divided into groups; Control, HI, THC and HI + THC. Each group consists of 8 animals. HI and HI + THC groups were given 10% fructose in the drinking waterfor 12weeks. In the last four weeks of the experiment, 1.5mgkg-1 THC was injected intraperitoneally daily into THC and HI + THC groups. The expression of tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), and nuclear factor-kappa B (NF-κB) were detected. JNK/SAPK and Grap2/p38 levels, total antioxidant and oxidant capacities (TAC and TOC) were analyzed in the pancreas. Levels of IL-6, NF-κβ, and TNF-α mRNA expression were higher in the pancreas with HI than in the control (p < 0.001 for all). THC treatment reduced the expression of IL-6, NF-κβ, and TNF-α mRNAs in the HI + THC group compared to the HI group (p < 0.001 for all). TOC increased in the HI group compared to the control group (p < 0.001). However, THC treatment reduced TOC levels in the HI + THC group compared to the HI group (p < 0.001). According to the results, the THC treatment may regulate inflammation and TOC in rats with hyperinsulinemia. Thus, we can say that THC may have anti-inflammatory and antioxidant potential in metabolic disorders.

Dimethyl Fumarate Attenuates Di-(2-Ethylhexyl) Phthalate-Induced Nephrotoxicity Through the Nrf2/HO-1 and NF-κB Signaling Pathways.

This study aimed to clarify the nephroprotective effect of dimethyl fumarate (DMF) against Di (2-ethylhexyl) phthalate (DEHP)-induced nephrotoxicity in both in vitro and in vivo models. The HEK-293 cells were exposed to different concentrations of DMF plus IC50 concentration of monoethylhexyl phthalate (MEHP) (the main metabolite of DEHP). Then, some of the oxidative stress parameters including ROS, MDA, and GSH, and cytotoxicity (MTT assay) were determined in treated cells. For in vivo evaluation, rats were divided into 7 groups (n = 6 per group). Corn oil group (gavage), DEHP group (200mg/kg dissolved in corn oil, gavage), DMF (15, 30, and 60mg/kg, gavage) plus DEHP (200mg/kg) groups, DMF (60mg/kg, gavage) alone, and vitamin E (20mg/kg, intraperitoneal (IP)) plus DEHP (200mg/kg) group. This treatment continued for 45days. Then, BUN and creatinine were evaluated by a commercial kit based on the urease enzymatic method and the Jaffe method, respectively. Mitochondrial oxidative stress and mitochondrial dysfunction parameters were evaluated using appropriate reagents, and gene expression of the p65 nuclear factor kappa B (NF-κB), tumor necrosis factor alpha (TNFα), nuclear factor E2-related factor 2 (Nrf2), and heme oxygenase-1 (HO-1) were evaluated by real-time PCR method. High concentrations of DMF significantly increased cell viability, and GSH content and significantly decreased ROS and MDA levels compared with the MEHP group in HEK-293 cells. DMF (60mg/kg) significantly decreased BUN and creatinine levels compared with the DEHP group. Mitochondrial function and mitochondrial swelling were significantly improved in DMF group (60mg/kg) compared with the DEHP group. DMF (30 and 60mg/kg) significantly improved MMP collapse compared with the DEHP group. DMF (30 and 60mg/kg) significantly decreased ROS levels compared with the DEHP group in isolated kidney mitochondria. DMF (60mg/kg) significantly decreased MDA levels and significantly increased GSH content compared with DEHP group in isolated kidney mitochondria. The mRNA expression levels of Nrf2 and HO-1 were significantly reduced in the DEHP group compared to the control group and were significantly increased in the DMF group compared to the DEHP group. p65NF-κB and TNFα mRNA expression levels were significantly increased in the DEHP group compared to the control group. However, DMF significantly decreased p65NF-κB and TNFα mRNA expression compared to the DEHP group. DMF can act as a nephroprotective agent against DEHP partly through modulation of oxidative stress, mitochondrial function, and inflammation.

Study on the Therapeutic Effect of Lenalidomide on Hemophilic Arthropathy

To explore the effect of lenalidomide on human fibroblast-like synovial cells (HFLS) and the therapeutic efficacy on hemophilic arthropathy in hemophilia A mice model. In vitro, to remodel the inflammatory environment of synovial tissue after hemorrhage, ferric citrate and recombinant TNF-α were added into the cell culture medium of HFLS. Cell Counting Kit-8 (CCK-8), Enzyme-linked immunosorbent assay (ELISA), Quantitative Real-time PCR (RT-qPCR) and flow cytometry were employed for detection of the effects of lenalidomide on the proliferation ability, pro-inflammatory cytokines release and apoptosis of HFLS cells. In vivo, hemophilia arthropathy was remodeled in hemophilia A mice by induction of hemarthrosis. A series of doses of lenalidomide (0.1, 0.3 and 1.0 g/kg) was administrated intra-articularly. Tissues of knee joints were collected on the 14th day after administration, and the protective effect of lenalidomide on arthritis in hemophilia A mice were evaluated by RT-qPCR and histological grading. In vitro, compared with the untreated control group, lenalidomide could significantly inhibit the proliferation of HFLS cells (P<0.05), and the effect was the most significant when the concentration was 0.01 μmol/L (P<0.001). Compared with the control group, lenalidomide could significantly inhibit the expression levels of TNF-α, IL-1β, IL-6 and IFN-γ in HFLS cells (P<0.05). The flow cytometry results showed that lenalidomide could enhance the apoptotis of HFLS cells (P<0.05). The results of RT-qPCR showed that lenalidomide could significantly reduce the mRNA expression levels of TNF-α, IL-1β, IL-6,MCP-1 and VEGF in the joint tissues (P<0.05). Histological results showed that compared with the injured group, lenalidomide could significantly reduce the pathological sequela after hemarthrosis induction, e.g. synovial thickening and neo-angiogenesis in the synovium. The protection displayed a dose-response pattern roughly. In vitro, lenalidomide can inhibit the proliferation of HFLS cells, promote their apoptosis, and inhibit the expression of pro-inflammatory cytokines. In vivo, lenalidomide can significantly decrease the expression of pro-inflammatory cytokines in the joints of mice, and prevent the development of inflammation and neo-angiogenesis. The results provide a theoretical and experimental basis for the clinical application of lenalidomide in the treatment of hemophilic arthropathy.

Ginsenoside Rb1 attenuates lipopolysaccharide-induced chronic neuroinflammation in mice by tuning glial cell polarization

To evaluate whether ginsenoside Rb1 (Rb1) can attenuate lipopolysaccharide (LPS)-induced chronic neuroinflammation in mice and to explore its relationship with glial cell polarization. Intraperitoneal injection with an escalating dose of LPS was used to establish a chronic neuroinflammation model in mice. Once LPS was initiated, 10 or 20 mg/kg Rb1, or sterile saline, was administered for 14 consecutive days. Open field test and beam walking test were used to monitor the changes in behavior. The concentration of cytokines in the serum and brain were used to monitor the systemic inflammation and neuroinflammation, respectively. Molecules specific to each glial cell phenotype were used to investigate glial cell polarization. Mice in the LPS group had reduced spontaneous activities and impaired beam walking performance. Rb1 obviously eased LPS-induced behavior disturbances. Regarding the levels of serum cytokines, both tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) were significantly increased, while interleukin-10 (IL-10) and transforming growth factor β (TGF-β) remarkably decreased after LPS treatment (all P < .001). Rb1 treatment significantly attenuated LPS-induced serum cytokine changes (all P < .05). The results of quantitative polymerase chain reaction and western blotting showed that the mRNA and protein expression levels of TNF-α and complement component 3 (C3) in the brain were significantly increased after LPS treatment (all P < .01). Rb1 treatment significantly inhibited LPS-induced inflammation in the brain (all P < .05). Glial cell polarization analysis showed that M1 and M2 microglia, and A1 astrocytes increased following LPS treatment, while A2 astrocytes decreased. Rb1 treatment reduced M1 and M2 microglia, and A1 astrocytes, and significantly increased A2 astrocytes. Rb1 can attenuate chronic neuroinflammation induced by LPS in mice, which may be partially attributable to its fine tuning of microglia and astrocyte polarization. Rb1 has potential value for treating neurodegenerative diseases.

Piperine Derived from Piper nigrum L. Inhibits LPS-Induced Inflammatory through the MAPK and NF-κB Signalling Pathways in RAW264.7 Cells.

Piperine, an important natural product, has a good anti-inflammatory effect. However, few researchers have studied its mechanism in these pathways. The objective of this research was to evaluate the molecular mechanism underlying the anti-inflammatory responses of piperine in lipopolysaccharide (LPS)-stimulated RAW264.7 cells. The purification and characterization of piperine from Piper nigrum L. were determined by HPLC, UPLC-Q-TOF-MS and 1H NMR. Then, the anti-inflammatory activity was evaluated by a reagent test kit, ELISA kits, RT-PCR and Western blot experiments. The results suggested that piperine (90.65 ± 0.46% purity) at a concentration of 10–20 mg/L attenuated the production of NO and ROS, downregulated the protein and mRNA expression levels of TNF-α, IL-1β and IL-6, and upregulated the protein and mRNA transcription levels of IL-10. Meanwhile, the Western blot results indicated that piperine could inhibit the phosphorylation levels of the ERK, JNK, p38 and p65 proteins. Our findings suggest that piperine is a potential anti-inflammatory substance, whose molecular mechanism may be to regulate the key factors of the NF-κB and MAPK signalling pathways.

Neospora caninum inhibits tumor development by activating the immune response and destroying tumor cells in a B16F10 melanoma model

BackgroundMelanoma is a malignant tumor with a high mortality rate. Some microorganisms have been shown to activate the immune system and limit cancer progression. The objective of this study is to evaluate the anti-melanoma effect of Neospora caninum, a livestock pathogen with no pathogenic activity in humans.MethodsNeospora caninum tachyzoites were inoculated into a C57BL/6 mouse melanoma model by intratumoral and distal subcutaneous injections. Tumor volumes were measured, and cell death areas were visualized by hematoxylin and eosin staining and quantified. Apoptosis in cell cultures and whole tumors was detected by propidium iodide (PI) and TUNEL staining, respectively. Cytokine and tumor-associated factor levels in tumors and spleens were detected by real-time quantitative polymerase chain reaction. Infiltration of macrophages and CD8+ T cells in the tumor microenvironment (TME) were detected by immunohistochemistry with anti-CD68 and anti-CD8 antibodies, respectively. Finally, 16S rRNA sequencing of mice cecal contents was performed to evaluate the effect of N. caninum on gut microbial diversity.ResultsIntratumoral and distal subcutaneous injections of N. caninum resulted in significant inhibition of tumor growth (P < 0.001), and more than 50% of tumor cells were dead without signs of apoptosis. Neospora caninum treatment significantly increased the mRNA expression levels of IL-12, IFN-γ, IL-2, IL-10, TNF-α, and PD-L1 in the TME, and IL-12 and IFN-γ in the spleen of tumor-bearing mice (P < 0.05). An increase in the infiltration of CD8+ T cells and macrophages in the TME was observed with these cytokine changes. Neospora caninum also restored the abundance of gut microbiota Lactobacillus, Lachnospiraceae, Adlercreutzia, and Prevotellaceae associated with tumor growth, but the changes were not significant.ConclusionNeospora caninum inhibits B16F10 melanoma by activating potent immune responses and directly destroying the cancer cells. The stable, non-toxic, and efficacious properties of N. caninum demonstrate the potential for its use as a cancer treatment.Graphical