mRNA Levels Of TNF-α Research Articles

Three water soluble polysaccharides with anti-inflammatory activities from Selaginella uncinata (Desv.) Spring

Three water soluble polysaccharides named SUSP-1, SUSP-2 and SUSP-3 from Selaginella uncinata (Desv.) Spring were purified, which contained different contents of galactose, arabinose, mannose, glucose and xylose, and SUSP-3 had large amount of galacturonic acid. Structural identification showed that the backbone structure of SUSP-1 was composed of (1 → 2)-α-D-Manp, (1 → 4)-α-D-Manp and (1 → 4)-β-D-Xylp. The main chains of SUSP-2 were (1 → 3)-α-D-Galp and (1 → 4)-α-D-Glcp, and SUSP-3 had two fragments and the main chains were (1 → 4)-α-D-GalpA and (1 → 4)-β-D-Xylp. Furthermore, their anti-inflammatory activities were evaluated. THP-1 monocytes were induced into macrophages by phorbol 12-myristate 13-acetat (PMA) and then stimulated by lipopolysaccharides (LPS). The data showed that compared with model groups, SUSP-1, SUSP-2 and SUSP-3 significantly inhibited ROS levels, promoted IL-10 expression, suppressed the mRNA levels of IL-6, TNF-α and IL-1β, and effectively blocked LPS binding to CD14 receptor to reduce inflammation. This study provided new data for the development of natural polysaccharides from S. uncinata with anti-inflammatory activities.

КОГНИТИВНЫЕ НАРУШЕНИЯ И СОСТОЯНИЕ ГЛИАЛЬНЫХ КЛЕТОК МОЗГА В ОТДАЛЕННЫЙ ПЕРИОД ПОСЛЕ ГАММА-ОБЛУЧЕНИЯ ГОЛОВЫ МЫШЕЙ

Purpose: To investigate the effect of fractionated whole-brain γ-irradiation at a cumulative dose of 20 Gy on cognitive functions, the state of brain glial cells and expression of multiple cytokines in mice 2 months after exposure. Material and methods: Male C57Bl/6 mice were exposed to fractionated head γ-irradiation with 5 doses of 4 Gy. Two months after irradiation the behavior and cognitive functions of animals were assessed. After isolation of cells from mice brains the content of resting and activated microglia, microglial cells with M1- and M2-phenotype, astrocytes, proliferating cells were evaluated, and the hippocampal mRNA levels of pro- and anti-inflammatory cytokines (TNFα, IL-1β, IL-6, IL-4, TGFβ) were determined. Results: It was shown that fractionated head γ-irradiation didn’t alter the locomotor activity and associative (context fear) memory, but reduced the episodic memory in novel object recognition test (discrimination index was 0.44 ± 0.08 и 0.02 ± 0.09 in control and irradiated groups, respectively) and spatial memory in Morris water maze (time in target quadrant was 46,8 ± 2,4 % and 37,4 ± 2,8 % in control and irradiated groups, respectively). Exposure of γ-radiation significantly reduced the brain contents of microglial cells (Iba1+) and astrocytes (GFAP+) with concurrent 2.5 times increase in proportion of activated microglia (from 2.0 ± 0.2 % in control to 4.9 ± 0.5 % in irradiated mice), changed the M1- / M2-microglia ratio and significantly decreased the number of proliferating cells (BrdU+) and proliferating microglial cells (BrdU+/Iba1+). An increase in mRNA level of pro-inflammatory cytokine TNFα, a decrease in mRNA level of anti-inflammatory cytokine TGFβ and concurrent increase in mRNA level of IL-4 were detected in hippocampus 2 months after irradiation. Conclusion: We show that fractionated head γ-irradiation at a cumulative dose of 20 Gy reduces the episodic and spatial memory in mice 2 months after exposure. Cognitive dysfunctions detected are associated with neuroinflammation characterized by increasing proportion of activated brain microglia and altered hippocampal pro- and anti-inflammatory cytokine profile.

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.

Dietary sodium butyrate administration alleviates high soybean meal-induced growth retardation and enteritis of orange-spotted groupers (Epinephelus coioides)

An 8-week feeding trial was conducted to investigate whether dietary sodium butyrate (SB) administration alleviates growth reduction and enteritis of orange-spotted grouper (Epinephelus coioides) caused by high soybean meal (SBM) feeding. The control diet (FM diet) was formulated to contain 48% protein and 11% fat. Soybean meal was used to replace 60% FM protein in FM diet to prepare a high SBM diet (HSBM diet). Sodium butyrate (SB) at 0.1%, 0.2%, and 0.3% were added to HSBM diets to prepare three diets. Triplicate groups of 30 groupers (initial weight: 33.0 ± 0.3 g) were fed one of the diets twice daily, to apparent satiety. HSBM diets had lowered growth rate and feed efficiency vs FM diets (P &amp;lt;0.05). Growth rate and feed efficiency were improved by dietary SB administration and were in a dose-dependent manner (P &amp;lt;0.05). A similar pattern to the growth rate was observed for plasma LDL-C and gut digestive activity of lipase, trypsin, and protease, but the opposite trend was observed for intestinal contents of D-lactic acid and endotoxin, in response to dietary SB inclusion levels (P &amp;gt;0.05). The muscular thickness in the middle and distal intestines in SB-treated diets were higher than that in HSBM diets (P &amp;lt;0.05). The mRNA levels of intestinal pro-inflammatory cytokines IL-8, IL-1β, IL-12 and TNF-α had a decreasing trend, and the mRNA level of intestinal anti-inflammatory cytokine TGF-β1 had the opposite trend, with increasing SB inclusion levels (P &amp;lt; 0.05). The above results indicate that dietary SB intervention could enhance growth and feed utilization of groupers with SBM-induced enteritis by promoting intestinal digestive enzyme activities, reducing mucosa permeability, maintaining the integrity of intestinal morphology and attenuating the intestinal inflammatory response.

Transcript levels of COX-2, TNF-α, IL-6 and IL-10 in chronic obstructive pulmonary disease: An association with smoking and severity

Chronic obstructive pulmonary disease (COPD) is characterized by inflammation. Tobacco smoking plays an important role in inflammation and subsequent disease progression. Therefore, inflammatory biomarkers (COX-2, TNF-α, IL-6 and IL-10 genes) expression levels were marked in patients with and without smoking habits and the severity of COPD. The severity of COPD, spirometry results and pack-years of smoking were assessed. The mRNA levels of COX-2, TNF-α, IL-6 and IL-10 in peripheral blood mononuclear cells were evaluated using reverse transcriptase-polymerase chain reaction. Relative fold expression and Pearson correlations were established among the group variables. Patients had a higher expression of COX-2 followed by TNF-α and IL-6 (p&lt;0.000). In contrast, IL-10 expression was decreased by half in patients compared to controls (p&lt;0.0001). COX-2 (10.37-fold), IL-6 (11.31-fold) and TNF-α (6.86-fold) were highly expressed in smokers (P &lt;0.0001). In contrast, IL-10 expression was dramatically lower in smokers than non-smokers (p&lt;0.0001). With increasing smoking pack-years, COX-2, TNF-α and IL-6 expression were significantly increased, but IL-10 gene expression was decreased (p&lt;0.05). COX-2, TNF-α and IL-6 gene expression progressively increased with severity grade, but IL-10 expression was dramatically reduced (p&lt;0.0001). COX-2 was positively correlated with TNF-α and IL-6 (r=0.39, p&lt;0.0001) whereas COX-2 was moderately correlated with IL-10 (r=-0.62 and 0.63, p&lt;0.0001). The higher expression of COX-2, TNF-α and IL-6 in patients with COPD suggests that these patients will likely benefit from COX-2, TNF-α and IL-6 blockers. Smoking cessation reduces the burden and severity of COPD.

An Encapsulated Organic Acid and Essential Oil Mixture Improves the Intestinal Health of Weaned Piglets by Altering Intestinal Inflammation and Antioxidative Capacity.

Simple SummaryWeaning stress causes retarded growth, gut disorder and dysfunction, severe diarrhea and higher mortality in weaned piglets. Antibiotic growth promoters (AGP) have been conventionally used to alleviate the negative effects of weaning stress. However, the long-term use of AGP leads to various adverse effects, such as antimicrobial resistance and food drug residues, and threatens public safety. Therefore, AGP should be replaced with residue-free, pollution-free and toxin-free alternatives because several countries and regions have banned the use of AGP in the feed industry. This study investigates the effects of an encapsulated organic acid and essential oil mixture (OAEO) on the growth performance, immuno-antioxidant capacity and intestinal health of weaned piglets. The results reveal that OAEO as an alternative to AGP improved the growth performance, immuno-antioxidant status and intestinal health of weaned piglets partly by activating the Nrf2 signaling pathway and suppressing the TLR4/NF-κB signaling pathway.This study investigates the effects of an encapsulated organic acid and essential oil mixture (OAEO) on the growth performance, immuno-antioxidant capacity and intestinal health of weaned piglets. In total, 120 weaned piglets (23 days of age; 6.96 ± 0.08 kg) were randomly allotted to four treatments (six replicates/group; five piglets/replicate): the control group (CON) was fed the basal diet (BD), the antibiotic growth promoters group (AGP) received the BD with 20 mg/kg colistin sulphate and 10 mg/kg bacitracin zinc, and OAEO1 and OAEO2 were fed the BD with 1000 mg/kg and 2000 mg/kg OAEO, respectively. The trial lasted 21 days and then one piglet per replicate was selected for sample collection. OAEO increased the average daily gain, spleen index, serum interleukin (IL)-10, immunoglobulin (Ig) G and IgA levels; serum superoxide dismutase and glutathione peroxidase (GPX) activities; and jejunal villus height (VH), VH/crypt depth, goblet cell number, and amylase and trypsin activities (p < 0.05) compared with CON but reduced the diarrhea rate, serum tumor necrosis factor (TNF)-α, malondialdehyde (MDA), and D-lactic acid contents and diamine oxidase (DAO) activity (p < 0.05). OAEO also increased the jejunal zonula occludens-1, occludin, claudin-1, mucin-2, nuclear factor erythroid 2-related factor 2 (Nrf2), GPX and IL-10 mRNA levels, GPX activity and IL-10 content (p < 0.05) compared with CON but reduced jejunal MDA, IL-1β and TNF-α contents and Toll-like receptor (TLR) 4, nuclear factor (NF)-κB and TNF-α mRNA levels (p < 0.05). In addition, AGP increased ADG, serum IgA level and GPX activity, jejunal trypsin activity and IL-10 content and mRNA level (p < 0.05) compared with CON but reduced the serum TNF-α content and DAO activity and jejunal NF-κB mRNA level (p < 0.05). Overall, OAEO as an alternative to AGP improved the growth performance, immuno-antioxidant status and gut health of weaned piglets partly via activating the Nrf2 signaling pathway and suppressing the TLR4/NF-κB signaling pathway.

Ponciri Fructus Immaturus ethanol extract attenuates septic shock through inhibition of the STAT1 signaling pathway.

Sepsis is a systemic inflammatory disease to infections and results in tissue damage and multiple organ failure. Ponciri Fructus Immaturus (PFI) is widely used in traditional medicine for allergic inflammation and gastrointestinal disorders. However, the effect of PFI on sepsis is still unknown. This study investigated the anti-inflammatory and antiseptic effects of PFI ethanol extract (PFIE) in LPS-stimulated J774 macrophages and mice with CLP- or LPS-induced sepsis, respectively. PFIE attenuates the LPS-induced production of the proinflammatory mediator NO by inhibiting the expression of iNOS in J774 cells. Real-time RT-PCR data and ELISA showed that the mRNA and protein levels of TNF-α, IL-1β, and IL-6 increased in LPS-stimulated J774 cells. However, this induction was significantly suppressed in PFIE pre-treated J774 cells. We also found that PFIE administration increased the survival rate of mice with LPS- and CLP-induced sepsis. Decreased serum levels of AST, ALT, and CK were observed after administration of PFIE, which was associated with reduced production of proinflammatory factors, such as NO, TNF-α, IL-1β, and IL-6. Moreover, PFIE suppressed the phosphorylation and nuclear translocation of STAT1 in LPS-stimulated J774 cells, suggesting that PFIE can inhibit LPS- and CLP-induced septic shock by suppressing the STAT1 activation. These findings provide the potential therapeutic relevance of PFIE in treating acute inflammatory disease.

α-Asarone attenuates chronic sciatica by inhibiting peripheral sensitization and promoting neural repair.

This study explored the therapeutic effect of α-asarone on chronic sciatica. Thirty-two Sprague-Dawley (SD) rats were divided into four groups: the sham group, chronic constriction injury (CCI) group, pregabalin group, and α-asarone group. Hot hyperalgesia was induced after the CCI operation, and α-asarone was found to relieve chronic neuralgia. Furthermore, α-asarone reduced IL1β, IL6, TNF-α, CRP, and LPS levels and increased IL10 levels in serum. α-Asarone decreased the protein levels of TRPA1, TRPM8, and TRPV1-4 and the mRNA levels of TRPA1, TRPM8, TRPV1-4, IL1β, and TNF-α in dorsal root ganglion neurons. In the sciatic nerve, α-asarone treatment reduced the number of inflammatory cells and promoted the proliferation of Schwann cells, favouring recovery of the nerve structure. In cellular experiments, LPS induced Schwann cell apoptosis via TLR4/p38MAPK signalling; α-asarone attenuated LPS-induced Schwann cell apoptosis by decreasing TLR4, p-p38MAPK, cleaved-caspase3, and cleaved-caspase7 levels and increasing Bcl-2 and Bcl-xl expression. Overall, these findings suggest that α-asarone relieves chronic sciatica by decreasing the levels of inflammatory factors, inhibiting peripheral sensitization, and favouring the repair of damaged nerves.

Ameliorative Effect of Pomegranate Peel Extract (PPE) on Hepatotoxicity Prompted by Iron Oxide Nanoparticles (Fe2O3-NPs) in Mice.

An evaluation of the ameliorative effect of pomegranate peel extract (PPE) in counteracting the toxicity of iron oxide nanoparticles (Fe2O3-NPs) that cause hepatic tissue damage is focused on herein. Forty male albino mice were haphazardly grouped into four groups as follows: the first control group was orally gavage daily with physiological saline; the second group received 100 mg/kg of PPE by the oral route day after day; the third group received 30 mg/kg Fe2O3-NPs orally; and the fourth group received both PPE and Fe2O3-NPs by the oral route, the same as the second and third sets. Later, after the completion of the experiment, we collected the liver, blood, and bone marrow of bone specimens that were obtained for further laboratory tests. For instance, exposure to Fe2O3-NPs significantly altered serum antioxidant biomarkers by decreasing the levels of total antioxidant capacity (TAC), catalase (CAT), and glutathione s-transferase (GST). Additionally, it caused changes in the morphology of hepatocytes, hepatic sinusoids, and inflammatory Kupffer cells. Furthermore, they significantly elevated the number of chromosomal aberrations including gaps, breaks, deletions, fragments, polyploidies, and ring chromosomes. Moreover, they caused a significant overexpression of TIMP-1, TNF-α, and BAX mRNA levels. Finally, the use of PPE alleviates the toxicity of Fe2O3-NPs that were induced in the hepatic tissues of mice. It is concluded that PPE extract has mitigative roles against the damage induced by Fe2O3-NPs, as it serves as an antioxidant and hepatoprotective agent. The use of PPE as a modulator of Fe2O3-NPs’ hepatotoxicity could be considered as a pioneering method in the use of phytochemicals against the toxicity of nanoparticles.

Connexin 43 (Cx43) regulates high-glucose-induced retinal endothelial cell angiogenesis and retinal neovascularization.

Diabetic retinopathy (DR) is an important microvascular complication of type 1 and type 2 diabetes mellitus (DM) and a major cause of blindness. Retinal neovascularization plays a critical role in the proliferative DR. In this study, high glucose-induced connexin 43 (Cx43) expression in human retinal endothelial cells (hRECs) in a dose-dependent manner. Compared with hRECs under normal culture conditions, high-glucose (HG)-stimulated hRECs showed promoted tubule formation, increased ROS release, and elevated levels of tumor necrosis factor-alpha (TNF-α), interleukin-1β (IL-1β), vascular endothelial growth factor A (VEGFA), and intercellular adhesion molecule 1 (ICAM-1) in the culture medium. HG-induced alterations were further magnified after Cx43 overexpression, whereas partially eliminated after Cx43 knockdown. Finally, in the DR mouse model, impaired retinal structure, increased CD31 expression, and elevated mRNA levels of TNF-α, IL-1β, VEGFA, and ICAM-1 were observed; in-vivo Cx43 knockdown partially reversed these phenomena. Conclusively, Cx43 knockdown could inhibit hREC angiogenesis, therefore improving DR in the mouse model.

Abstract 061: Renal A20 Protects Against The Cisplatin-induced Kidney Injury

We recently reported that the ubiquitin-editing protein A20 in myeloid cells can limit the severity of hypertension. However, whether A20 in the kidney regulates renal injury without impacting systemic immune responses remains unclear. To address this question, we bred A20 flox/flox mice with the Pax8-rtTA and Tet-On lines, generating inducible renal epithelial cell A20 knockout mice (A20 iKKO). Mice with all 3 transgenes were used as the A20 iKKO group, whereas mice lacking the Pax8-rtTA or Tet-On transgene were wild-type (WT) controls. Before the experiments, mice were given 2mg/ml of doxycycline in water containing 5% (5 of 100) sucrose for 2 weeks so that A20 in renal tubular cells was ablated. 3 days after a single dose of cisplatin (20mg/kg), A20 iKKO mice exhibited more severe kidney injury compared to WTs (blinded injury score, 3.2±0.41 vs. 2.1±0.35 au, p=0.039). A20 iKKOs also had upregulated renal mRNA levels for NGAL (6.0±1.91 vs. 1.0±0.18 au, p=0.018) and KIM-1(3.3±1.2 vs. 1.0±0.29 au, p=0.07). Injured kidneys from the A20 iKKOs contained greater absolute numbers of T cells (123.6±8.5 vs. 53.8±4.7, x10 4 cells per gram kidney, p&lt;0.001), macrophages (159.7±22.5 vs. 52.7±9.9, x10 4 cells per gram kidney, p&lt;0.001), and dendritic cells (66.7±10.6 vs. 32.6±6.1, x10 4 cells per gram kidney, p=0.015) than WTs by flow cytometric analysis. In turn, mRNA levels for TNF-α (9.1±2.62 vs. 1.0±0.18 au; p=0.006) and IL-1β (4.1±1.06 vs. 1.0±0.13 au; p=0.009) were increased in the injured A20 iKKO injured kidneys vs WTs. Thus, A20 in the kidney epithelium protects against cisplatin-induced kidney injury by constraining renal immune cell accumulation and inflammatory cytokine release.

Taxifolin attenuates inflammation via suppressing MAPK signal pathway in vitro and in silico analysis

ObjectiveTaxifolin is a natural flavonoid compound that can be isolated from onions, grapes, oranges and grapefruit. It also acts as a medicine food homology with extraordinary antioxidant and anti-inflammatory activity. This study aims to explain the protective effects and potential mechanisms of taxifolin against inflammatory reaction. MethodsLevels of interleukin (IL)-6, IL-1β and intracellular reactive oxygen species (ROS) were assessed in different time after the treatment of taxifolin in RAW264.7 cells induced by lipopolysaccharide (LPS). Subsequently, the mRNA and protein levels of inducible nitric oxide synthase (iNOS), vascular endothelial growth factor (VEGF), cyclooxygenase (COX)-2, tumor necrosis factor (TNF)-α and the phosphorylation expression levels of the MAPK signal pathway were also evaluated. A silico analysis was used to explain the binding situation for the investigation of taxifolin and MAPK signal pathway. And then MAPK inhibitors were used to reveal the expression level of iNOS, VEGF, COX-2 and TNF-α in RAW264.7 cells. ResultsIt was demonstrated that cell inflammatory damage induced by LPS was significantly alleviated after the treatment of taxifolin. Then, the mRNA and protein levels of iNOS, VEGF, COX-2 and TNF-α were reduced and the phosphorylation expression levels of the MAPK signal pathway were down-regulated remarkably as well. In silico analysis, taxifolin could form a relatively stable combination with MAPK signal pathway. MAPK inhibitors showed increasing or decreasing effect in the mRNA levels of iNOS, VEGF, COX-2 and TNF-α, which suggesting that taxifolin down-regulated iNOS, VEGF, COX-2 and TNF-α expressions were not entirely through the MAPK pathway. ConclusionThis finding demonstrated that taxifolin improved the inflammatory responses that partly involved in the phosphorylation expression level of MAPK signal pathway in RAW264.7 cells exposed to acute stress.

TNF-α and NF-κB signaling play a critical role in cigarette smoke-induced epithelial-mesenchymal transition of retinal pigment epithelial cells in proliferative vitreoretinopathy.

Proliferative vitreoretinopathy (PVR) is characterized by the growth and contraction of cellular membranes within the vitreous cavity and on both surfaces of the retina, resulting in recurrent retinal detachments and poor visual outcomes. Proinflammatory cytokines like tumor necrosis factor alpha (TNFα) have been associated with PVR and the epithelial-mesenchymal transition (EMT) of retinal pigment epithelial (RPE) cells. Cigarette smoke is the only known modifiable risk factor for PVR, but the mechanisms are unclear. The purpose of this study was to examine the impact of cigarette smoke on the proinflammatory TNFα/NF-κB/Snail pathway in RPE cells to better understand the mechanisms through which cigarette smoke increases the risk of PVR. Human ARPE-19 cells were exposed to cigarette smoke extract (CSE), for 4 to 24-hours and TNFα, Snail, IL-6, IL-8, and α-SMA levels were analyzed by qPCR and/or Western blot. The severity of PVR formation was assessed in a murine model of PVR after intravitreal injection of ARPE-19 cells pre-treated with CSE or not. Fundus imaging, OCT imaging, and histologic analysis 4 weeks after injection were used to examine PVR severity. ARPE-19 cells exposed to CSE expressed higher levels of TNFα, SNAIL, IL6 and IL8 mRNA as well as SNAIL, Vimentin and α-SMA protein. Inhibition of TNFα and NF-κB pathways blocked the effect of CSE. In vivo, intravitreal injection of ARPE-19 cells treated with CSE resulted in more severe PVR compared to mice injected with untreated RPE cells. These studies suggest that the TNFα pathway is involved in the mechanism whereby cigarette smoke increases PVR. Further investigation into the role of TNFα/NF-κB/Snail in driving PVR and pharmacological targeting of these pathways in disease are warranted.

Tanshinone IIA Ameliorates Nonalcoholic Steatohepatitis in Mice by Modulating Neutrophil Extracellular Traps and Hepatocyte Apoptosis.

Salvia miltiorrhiza Bunge, a traditional Chinese medicine, is widely used in the treatment of a variety of diseases and syndromes. Tanshinone IIA (TIIA), a phenanthrenequinone-class derivative extracted from S. miltiorrhiza, is one of its main active components and has anti-inflammatory effects on various tissues and cells. This study aimed to investigate the beneficial effects of TIIA on nonalcoholic steatohepatitis (NASH) induced in mice using a methionine choline deficiency (MCD) diet and the underlying mechanism of these. Our results reveal that TIIA remarkably ameliorated hepatic steatosis and inflammation and decreased the serum levels of liver dysfunction markers while increasing the levels of serum total cholesterol and triglycerides in MCD-fed mice. TIIA significantly reduced mRNA levels of the inflammatory factors TNF-α, IL-6, and TGF-β. Similarly, TIIA inhibited caspase-3 and Bax-mediated apoptosis in MCD-fed mice. Together, our data indicate that TIIA inhibits the formation of MPO and CitH3 in neutrophil extracellular traps and inhibits apoptosis mediated by caspase-3 and Bax in hepatocytes, thereby mitigating inflammatory progression in an MCD diet-induced NASH mouse model.

Abstract 003: Treatment With Ligilactobacillus Improves Gut Permeability And Hypertension In Spontaneously Hypertensive Rats

Emerging evidence provide that a microbial imbalance (dysbiosis) is linked to several diseases including cardiovascular diseases. One of the features in hypertension is increased gut permeability. Supporting this, plasma endotoxin level is increased in hypertension and our previous study indicated that the plasma level of exotoxin from Streptococcus was increased in spontaneously hypertensive rats (SHR). Thus, we hypothesized that improvement of gut permeability in hypertension might be expected to prevent development of hypertension. In this study, we performed a 16S metagenome analysis using a synthetic long read sequencing approach by LoopSeq in feces from 10- to 15-week old SHR and Wistar-Kyoto rat (WKY). The analysis revealed that two difference species of anti-inflammatory microbiome were strongly decreased in SHR compared with WKY (n=4). We focus on the decreased Ligilactobacillus sp. (LAB) in SHR and oral administration of LAB to SHR for 2 weeks. SHR (10-15-week-old) exhibited increased gut permeability in proximal colon (P&lt;0.05, n=4), which was improved by treatment with LAB for 2 weeks (P&lt;0.05, n=4). The expression level of tight junction protein Claudin4 was decreased in SHR proximal colon (SHR vs WKY, 1.0±0.1 vs 1.7±0.2-fold increase to SHR, P&lt;0.05, n=8), which was significantly increased by LAB treatment (SHR vs SHR+LAB, 1.0±0.1 vs 2.8±0.5-fold increase to SHR, P&lt;0.05, n=10). Proximal colon in SHR increased the expression level of TNF-α mRNA (1.0±0.2 vs 0.2±0.1, P&lt;0.05, n=8), which was reversed by LAB (1.0±0.2 vs 0.3±0.1, P&lt;0.05, n=10). The level of mucin 2 was lower in SHR proximal colon (1.0±0.2 vs 4.3±0.8, P&lt;0.05, n=8), which was not reversed by LAB (1.0±0.1 vs 0.8±0.3, n=10). We observed that high blood pressure in SHR was attenuated by treatment with LAB for 2 weeks (194.0±5.9 vs 149.7±4.8 mmHg, P&lt;0.05, n=10). LAB also improved decrease in acetylcholine-induced relaxation in SHR mesenteric artery (P&lt;0.05, n=5-8). We conclude that treatment of SHR with the decreased anti-inflammatory microbiota, LAB improves gut permeability in proximal colon possibly through inhibition of inflammation, resulting in lowering blood pressure. These results suggest that anti-inflammatory microbiome might be a new therapeutic target for hypertension.

Ferulic acid alleviates sciatica by inhibiting peripheral sensitization through the RhoA/p38MAPK signalling pathway

BackgroundNonsteroidal anti-inflammatory drugs are used to relieve sciatica, but their effects are not satisfactory. PurposeThis study aimed to explore the therapeutic effects of ferulic acid on sciatica. MethodsThirty-two SD rats were randomly divided into 4 groups, i.e., sham operation group, chronic constriction injury (CCI) group, mecobalamin group, and ferulic acid group. We conducted behavioural tests, ELISA, PCR, Western blots, and immunofluorescence analysis. Specific inhibitors were used in cell experiments to explore the related mechanisms. ResultsThermal hyperalgesia was induced after CCI operation, and ferulic acid relieved thermal hyperalgesia. In addition, ferulic acid decreased the IL1β, IL6, TNF-α, and CRP mRNA levels; the IBA-1, iNOS, IL1β, RhoA, RhoA-GTP, COX2, Rock1, TRPV1, TRPA1, and p-p38MAPK levels in dorsal root ganglion (DRG) neurons; and the LPS, CRP, substance P (SP), and prostaglandin E2 (PGE2) levels in serum, and these levels were higher in the CCI group. In the cell experiments, LPS induced M1 polarization of GMI-R1 cells via the RhoA/Rock pathway. Ferulic acid attenuated LPS-induced M1 polarization by decreasing the levels of M1 polarization markers, including IL1β, IL6, TNF-α, iNOS, and CD32, and increased M2 polarization by increasing the levels of M2 polarization markers, including CD206 and Arg-1. LPS treatment clearly increased the iNOS, IL1β, RhoA, Rock1, Rock2 and p-p38 MAPK levels and reduced Arg-1 expression, and ferulic acid reversed these changes. ConclusionFerulic acid can inhibit peripheral sensitization by reducing the levels of inflammatory factors, TRPA1 and TRPV1 through the RhoA/p38 MAPK pathway to alleviate sciatica.

Paeonol alleviates placental inflammation and apoptosis in preeclampsia by inhibiting the JAK2/STAT3 signaling pathway.

Preeclampsia (PE) is a multisystemic and placental inflammatory disease that causes maternal and infant health issues. As one of the active components in peony root extract, paeonol (Pae) exerts anti-apoptosis and anti-inflammatory effects. Nonetheless, the protective role of Pae in PE has not yet been characterized. A mouse model of PE was constructed through tail vein injection of 1mg/d phosphatidylserine/dioleoyl-phosphatidycholine suspension. The levels of inflammatory cytokines in the placenta were examined via enzyme-linked immunosorbent assay (ELISA). The mRNA levels of inflammatory cytokines (TNF-α, IL-6, IFN-γ, and IL-4) and apoptosis markers (Bax, Bcl-2, and caspase-3) were tested using quantitative reverse transcription-polymerase chain reaction (qRT-PCR). Western blot analysis was performed to detect the protein levels of apoptosis markers and Janus kinase 2/signal transducer and activator of transcription 3 (JAK2/STAT3) signaling pathway-related molecules. Here, Pae repressed the inflammatory response in the placenta of PE-like mouse models, as demonstrated by the decreased concentrations and mRNA levels of TNF-α, IL-6, and IFN-γ and the increased concentrations and mRNA levels of IL-4. Apoptosis in the placentas of PE-like mouse models was attenuated by Pae, as manifested by the downregulated mRNA and protein levels of Bax and cleaved-caspase-3 and the upregulated Bcl-2. Administration of Pae inhibited the phosphorylation of JAK2 and STAT3 in the placental tissues of PE mice. The JAK2/STAT3 pathway agonist (SC-39100) reversed Pae treatment-mediated suppression of placental inflammation and apoptosis in PE mice. Overall, Pae inhibits the JAK2/STAT3 signaling pathway to attenuate placental inflammation and apoptosis in PE mice.

Caspase-11/4 is involved in bacteria-mediated periodontitis by promoting the release of interleukin-1 β and tumor necrosis factor-α

ObjectiveThis study investigated the main mechanism and role of caspase-11/4 as a pattern recognition receptor (PRR) in periodontitis through caspase-11 inhibition. DesignClinical tissue samples were collected from patients with periodontitis and healthy volunteers and evaluated through hematoxylin–eosin (HE) staining, immunohistochemical (IHC) staining, and real-time quantitative PCR (RT-qPCR). In the rat periodontitis model, both these staining procedures, RT-qPCR, and western blotting were used to evaluate the histological, mRNA, and protein levels of caspase-11, interleukin-1β (IL-1β), and tumor necrosis factor-α (TNF-α). In vitro, the role of caspase-11, inhibited by siRNA, was investigated by analyzing the mRNA and protein levels of IL-1β and TNF-α in Porphylinomonas gingivalis (P. gingivalis) lipopolysaccharide (LPS)-stimulated Raw264.7 macrophages. ResultsHistological and molecular biological results of clinical and experimental animal periodontitis samples indicated that caspase-11/4 mRNA and protein levels significantly increased in inflammatory tissues. Caspase-11 is mainly distributed in leukocytes, which are labeled by CD45 in the submucosa. In vitro results further confirmed that the expression of caspase-11/4, IL-1β, and TNF-α significantly increased in LPS-stimulated macrophages, and these changes were significantly attenuated by inhibiting caspase-11/4 expression. ConclusionsThe function of caspase-11 in rat periodontitis models is similar to that of caspase-4 in human clinical periodontitis. IL-1β and TNF-α release in periodontitis depends on the recognition of P. gingivalis LPS by caspase-11/4.

Study of the mechanism by which MSCs combined with LITUS treatment improve cognitive dysfunction caused by traumatic brain injury

Traumatic brain injury (TBI) substantially affects the quality of life of patients, and an effective therapy is unavailable. Previous studies have shown that mesenchymal stem cells (MSCs) and low-intensity transcranial ultrasound (LITUS) are effective treatments for neurological damage, inflammation, edema and cognitive impairment caused by TBI. However, it is unclear whether the combination of the two treatments exerts an additive effect. In this study, a rat TBI model was established using the controlled cortical impact (CCI) method. Neurological function was assessed by determining the rat modified neurological score (mNSS), and cognitive function was assessed using the Y-maze. Pathological changes in the injured tissue were observed using hematoxylin-eosin (HE) staining and immunohistochemistry (IHC), and western blot was performed to detect the expression levels of Nestin, neuron-specific enolase (NSE), glial fibrillary acidic protein (GFAP), growth-associated protein-43 (GAP-43), postsynaptic density protein (PSD-95), brain-derived neurotrophic factor (BDNF), tumor necrosis factor-α (TNF-α), and aquaporin-4 (AQP-4). Real-time fluorescence quantitative polymerase chain reaction (RT–PCR) was performed to detect the expression levels of GAP-43, PSD-95, BDNF, TNF-α, and AQP-4 mRNA to investigate whether MSCs combined with LITUS exert an additive therapeutic effect of alleviating the cognitive dysfunction caused by TBI and the possible mechanisms involved. Rats exhibited cognitive dysfunction 28 days after TBI, and MSCs combined with LITUS treatment ameliorated the cognitive deficits caused by TBI via increasing Nestin, NSE, GAP-43, PSD-95, and BDNF expression and attenuating the inflammatory response and edema caused by TBI via reducing TNF-α and AQP-4 expression. According to these results, MSCs combined with LITUS is more effective than MSCs alone for the treatment of TBI, and the mechanism may be the promotion of neuronal proliferation and differentiation, and the attenuation of the inflammatory response and edema, which ameliorates the spatial learning memory impairment caused by TBI. MSCs combined with LITUS treatment represents a new approach for the clinical treatment of patients with TBI.

Propofol directly binds to and inhibits TLR7.

Sedatives/anesthetics are important medical tools to facilitate medical care and increase patients' comfort. Increasingly, there is recognition that sedatives/anesthetics can modulate immune functions. Toll-like receptors (TLRs) are major pattern recognition receptors involved in the recognition of microbial components. TLR7 recognizes single-strand RNA virus such as influenza and SARS-CoV2 viruses and initiates interferon (IFN) responses. IFN production triggered by TLR7 stimulation is a critical anti-viral response. For example, patients with TLR7 variants including loss-of- function variants were associated with severe COVID-19. Taken together, it is important to determine if sedatives/anesthetics mitigate TLR7 function. We have previously showed that TLR7-mediated activation was not affected by volatile anesthetics. However, we found that propofol attenuated TLR7 activation among intravenous sedatives in the reporter assay. TLR7 agonist R837 stimulation increased TNF-α, IL-1β, IL-6, IL-10, and IFN-β mRNA levels in bone marrow-derived dendritic cells, while these levels were attenuated by propofol. Our murine lung slice experiments showed that propofol attenuated IFN production. R837 increased IFN-β expression in the lungs, and propofol attenuated IFN-β expression in an in vivo model of R837 intranasal instillation. We also found that propofol directly bound to and hindered its association of TLR7 with MyD88. Our analysis using fropofol, propofol derivative showed that the hydroxyl group in propofol was important for propofol-TLR7 interaction.