TNF-α In Mice Research Articles

CB2R agonist GW405833 alleviates acute liver failure in mice via inhibiting HIF-1α-mediated reprogramming of glycometabolism and macrophage proliferation.

The inflammatory responses involving infiltration and activation of liver macrophages play a vital role in acute liver failure (ALF). In the liver of ALF mice, cannabinoid receptor 2 (CB2R) is significantly upregulated on macrophages, while CB2R agonist GW405833 (GW) could protect against cell death in acute liver damage. In this study, we investigated the molecular mechanisms underlying the protective effects of GW against ALF in vivo and in vitro from a perspective of macrophage glycometabolism. Mice were pretreated with GW (10 mg/kg, i.p.), then were injected with D-GalN (750 mg/kg, i.p.) and LPS (10 mg/kg, i.p.) to induce ALF. We verified the protective effects of GW pretreatment in ALF mice. Furthermore, GW pretreatment significantly reduced liver macrophage infiltration and M1 polarization, and inhibited the release of inflammatory factors TNF-α and IL-1β in ALF mice. These protective effects were eliminated by CB2R antagonist SR144528 or in CB2R-/- ALF mice. We used LPS-stimulated RAW264.7 cells as an in vitro M1 macrophage-centered model of inflammatory response, and demonstrated that pretreatment with GW (10 μM) significantly reduced glucose metabolism by inhibiting glycolysis, which inhibited LPS-induced macrophage proliferation and inflammatory cytokines release. We verified these results in a stable CB2R-/- RAW264.7 cell line. Moreover, we found that GW significantly inhibited the expression of hypoxia inducible factor 1α (HIF-1α). Using a stable HIF-1α-/- RAW264.7 cell line, we confirmed that GW reduced the release of inflammatory cytokines from macrophages and inhibited glycolysis by downregulating HIF-1α expression. In conclusion, activation of CB2Rs inhibits the proliferation of hepatic macrophages and release of inflammatory factors in ALF mice through downregulating HIF-1α to inhibit glycolysis.

Structural characterization of a polysaccharide from Lyophyllum decastes with MAPK-mediated immune regulation ability in mice

Lyophyllum decastes is a common macrofungi for both medicinal and economic value, and its polysaccharide has excellent development potential as a new immune regulator. In the present study, the bioactive component polysaccharide of L decastes was extracted, and its chain structure was characterized in detail by Fourier transform infrared (FT-IR) spectroscopy, high-performance gel-permeation chromatography (HP-GPC), and gas chromatography–mass spectrometry (GC–MS). LDP-W had a molecular weight of 2.12 × 104 Da, highly branched β type pyranose. At the same time, it exhibited potential immunomodulatory activity, which can modulate the numbers of CD3+, CD4+, CD8+ and CD19+ cells in the spleen; optimize the CD4+/CD8+ ratio; promoted T and B lymphocyte proliferation and it can elevate the levels of IL-2, IL-6, IFN-γ and TNF-α in immunosuppressed mice by influencing the phosphorylation of MAPK signaling-related proteins. These studies fill the gap in the immune regulation mechanism of L decastes polysaccharide, providing a theoretical basis for the potential application of LDP-W as immunomodulatory drugs or functional foods.

Baitouweng decoction alleviates dextran sulfate sodium-induced ulcerative colitis by suppressing leucine-related mTORC1 signaling and reducing oxidative stress

Ethnopharmacological relevanceBaitouweng decoction (BTW) has been used for hundreds of years to treat ulcerative colitis (UC) in China and has produced remarkable clinical results. However, the knowledge in protective mechanism of BTW against UC is still unclear. Aim of the studyThe present study was designed to investigate the anti-UC effects of BTW and the underlying mechanisms involved. Methods3.5% dextran sulfate sodium (DSS)-induced experimental colitis was used to simulate human UC and the mice were treated with BTW (6.83 g/kg), leucine (200 mg/kg, Leu) or rapamycin (2 mg/kg, RAPA) as a positive control for 7 days. The clinical symptoms, serum myeloperoxidase (MPO) and malondialdehyde (MDA) levels were evaluated. Biological samples were collected to detect the effects of BTW on mechanistic target of rapamycin complex 1 (mTORC1) pathway and Leu metabolism. ResultsIn our study, BTW notably improved the clinical symptoms and histopathological tissue damage and reduced the release of proinflammatory cytokines, including IL-6, IL-1β and TNF-α in UC mice. BTW also alleviated oxidative stress by decreasing serum MPO and MDA levels. Additionally, BTW significantly suppressed mTORC1 activity in the colon tissues of UC mice. Serum metabolomics analysis revealed that the mice receiving BTW had lower Leu levels, which was in line with the decreased expression of branched-chain α-keto acid dehydrogenase kinase (BCKDK) in the colon tissues. Furthermore, oral administration of Leu aggravated DSS-induced acute colitis and enhanced mTORC1 activity in the colon. ConclusionThese data strongly demonstrated that BTW could ameliorate DSS-induced UC by regulating the Leu-related mTORC1 pathway and reducing oxidative stress.

Protective effect of rosmarinic acid-rich extract from Trichodesma khasianum Clarke against microbiota dysbiosis in high-fat diet-fed obese mice

Hypertrophy of adipose tissues and dysbiosis are hallmarks of obesity. Although drugs are applied for obesity treatment, side effects limit their use. The anti-obesity capacity of rosmarinic acid (RA) has been documented. Trichodesma khasianum Clarke is an edible RA-rich plant grown in Taiwan. Our previous study found that an 80 % ethanol extract of T. khasianum Clarke leaves (80EETC) ameliorates gastric mucosal damage through its anti-inflammatory, antioxidant, and microbiota modulation abilities. However, the anti-obesity effect of 80EETC remains unclear. Therefore, the objective of this study was to explore the protective effects of low-dose 80EETC (125 mg/kg b.w., 80EETCL) or high-dose 80EETC (250 mg/kg b.w., 80EETCH) on obesity development through gut microbiota modulation in high-fat diet (HFD)-induced C57BL/6 mice. The results showed a high RA content (89.2 ± 7.4 mg/g) in 80EETC. 80EETC administration significantly decreased body weight, body fat ratio, serum lipid levels (TC, TG, and LDL-C), adipose tissue accumulation, malondialdehyde (MDA), and tumor necrosis factor-α (TNF-α) in HFD-fed mice. Furthermore, supplementation with 80EETC reduced the Firmicutes/Bacteroidetes ratio and enhanced the relative abundance of gut microbiota (p_Bacteroidetes, f_Lactobacillus, f_Muribaculaceae, f_Prevotellaceae, g_Lactobacillus, g_Prevotellaceae_NK3B31_group, g_Ruminococcaceae_UCG–013, and g_Ruminococcaceae_UCG–014), which negatively correlated with obesity-related factors such as body weight, energy intake, fat accumulation in adipose tissue, TC, TG, LDL, and MDA. In conclusion, RA-rich 80EETC had a protective effect against obesity development and it has potential in healthy food applications.

Inhibition of the TLR/NF-κB Signaling Pathway and Improvement of Autophagy Mediates Neuroprotective Effects of Plumbagin in Parkinson's Disease.

A naphthoquinone molecule known as plumbagin (PL), which has a wide range of pharmacological properties including antitumor, antioxidation, anti-inflammation, and neuroprotective effects, is extracted from the roots of the medicinal herb Plumbago zeylanica L. Plumbagin has been studied for its potential to treat Parkinson's disease (PD). However, its effectiveness and mechanism are still unknown. This study intends to evaluate plumbagin's effectiveness against PD in vitro and in vivo. Plumbagin partially repaired the loss of dopaminergic neurons in the nigral substantia nigra and the resulting behavioural impairment caused by MPTP or MPTP/probenecid in mice. Furthermore, plumbagin treatment significantly inhibited the TLR/NF-κB pathways. It reduced the TNF-α, IL-6, and IL-1β mRNA expression in PD mice induced by MPTP or MPTP/probenecid, which was consistent with the findings in the inflammatory model of BV2 cells induced by MPP+ or LPS. In addition, plumbagin treatment enhanced the microtubule-associated protein 1 light chain 3 beta (LC3) LC3-II/LC3-I levels while decreasing the p-mTOR and p62 protein accumulation in PD mice induced by MPTP or MPTP/probenecid, which was similar to the results obtained from the experiments in SH-SY5Y and PC12 cells induced by MPP+. Consequently, our results support the hypothesis that plumbagin, by promoting autophagy and inhibiting the activation of the TLR/NF-κB signaling pathway, is a promising treatment agent for treating Parkinson's disease (PD). However, to confirm plumbagin's anti-PD action more thoroughly, other animal and cell PD models must be used in future studies.

Huangshan Floral Mushroom Polysaccharide Ameliorates Dextran Sulfate Sodium-Induced Colitis in Mice by Modulating Th17/Treg Balance in a Gut Microbiota-Dependent Manner.

Ulcerative colitis (UC) is a common chronic recurrent inflammatory bowel disease. This study attempts to reveal the improvement mechanism of floral mushroom polysaccharide (FMPS) on UC from the perspective of coordinated interaction between intestinal microbes and intestinal helper T cell 17 (Th17)/regulatory T cell (Treg) balance. Dextran sulfate sodium (DSS)-induced colitis mice model is used for the experiment. The results suggest that FMPS up-regulated the expression of occludin, ZO-1, and MUC2, and down-regulated the secretion of TNF-α, IL-1β, and IL-6 in colitis mice. Importantly, FMPS restores intestinal Th17/Treg balance. Meanwhile, FMPS can regulate intestinal microorganisms and improve the level of short-chain fatty acids (SCFAs) in colitis mice. Intestinal microbial depletion and fecal microbiota transplantation (FMT) experiments reveal that FMPS ameliorated UC is mediated by intestinal microbiome. Flow cytometry further proves that FMPS restores intestinal Th17/Treg balance in a microbial-dependent manner. These results indicate that FMPS has the potential to improve UC, and its mechanism depends on the restoration of Th17/Treg balance mediated by intestinal microorganisms. Therefore, it is suggested that FMPS dietary supplement can be potentially used to intervene UC.

Effects of early electroacupuncture on the expression of Iba-1 and TNF-α in Parkinson's disease mice

To observe the effect of early electroacupuncture(EA) intervention on ionized calcium binding adapter molecule 1 (Iba-1), tyrosine hydroxylase (TH) and tumor necrosis factor-α (TNF-α) in Parkinson's disease (PD) mice, so as to explore its neuroinflammation mechanism in treating PD. A total of 24 male C57BL/6J mice (9 weeks old) were randomly divided into control, model and EA groups, with 8 mice in each group. The PD model was established by long-term low dose subcutaneous injection of rotenone. Started at the same time with modeling, EA (2 Hz/100 Hz, 1 mA) was applied to "Shenting"(GV24), bilateral "Tianshu"(LI11), "Quchi"(ST25), and "Shangjuxu"(ST37) for 15 min, once a day for 8 weeks. The motor function was assessed by rotorod test and step length test. The expression levels of Iba-1 and TH proteins in substantia nigra pars compacta (SNpc) was detected by Western blot and immunohistochemistry. The expression level of TNF-α protein in colon tissue was examined by Western blot and immunofluorescence staining. Compared with the control group, the fall latency shortened at 4, 6, and 8 weeks after modeling (P<0.01) and the step length of hind limbs shortened at 5 and 7 weeks after modeling (P<0.01), the expression levels of Iba-1 in SNpc and TNF-α in colon tissue were increased (P<0.01), and the expression level of TH in SNpc was decreased (P<0.01) in the model group. Compared with the model group, the fall latency prolonged at 6 and 8 weeks after modeling (P<0.01) and the step length of hind limbs prolonged at 5 and 7 weeks after modeling (P<0.01), the expression levels of Iba-1 in SNpc and TNF-α in colon tissue were decreased (P<0.01, P<0.05), and the expression level of TH in SNpc was increased (P<0.05, P<0.01) in the EA group. Early EA intervention can delay the occurring time of motor disfunction, alleviated the loss of substantia nigra dopaminergic neurons, and exerted a good neuroprotective effect on the degenerative changes in rotenone-induced PD mice, which may be related to its effects in alleviating the intestinal inflammation, inhibiting the activation of microglia, thereby reducing the neuroinflammation.

Endothelin receptor B enhances liver injury and pro-inflammatory responses by increasing G-protein-coupled receptor kinase-2 expression in primary biliary cholangitis

Severe diseases like cirrhosis and liver failure can be developed from primary biliary cholangitis (PBC). Endothelin-2 (EDN2) and endothelin receptor B (EDNRB) are related to the pathogenesis of PBC. However, the roles of EDN2 and EDNRB in PBC-related liver injury and inflammation along with molecular mechanisms are poorly defined. In this study, histopathologic alterations of liver tissues were assessed through hematoxylin–eosin staining. Alanine transaminase (ALT), alkaline phosphatase (ALP), aspartate transaminase (AST), and γ-Glutamyltranspetidase (GGT) (4 liver function indexes) serum levels were detected with corresponding activity assay kits. Also, we determined the levels of M2 subtype anti-mitochondrial antibody (AMA-M2), interferon-gamma (IFN-γ), and tumor-necrosis factor alpha (TNFα) in serum with ELISA assay. Later, RT-qPCR assay was used to measure the expression of genes at mRNA levels, while western blotting and immunohistochemical techniques were used to detect protein levels of genes. Our results showed that the liver tissues of PBC patients and mice presented with severe hepatocyte injury and inflammatory cell infiltration as well as destruction of intrahepatic small bile ducts. ALP, AST, ALT, GGT, AMA-M2, IFN-γ, and TNF-α serum levels were higher in PBC patients and mice. Besides, EDN2 and EDNRB were highly expressed in serums and livers of PBC patients and mice. EDNRB potentiated PBC-related liver injury and pro-inflammatory responses, as evidenced by observation of serious liver pathologic injury and increased serum levels of ALP, AST, ALT, AMA-M2, IFN-γ, and TNF-α in PBC mice following EDNRB overexpression. EDNRB overexpression or activation via its agonist IRL-1620 TFA triggered liver injury and pro-inflammatory responses, increased GRK2 expression and induced NF-κB expression and activation in wild-type mice. EDNRB knockdown or inhibition by Bosentan alleviated liver damage and inflammation, reduced GRK2 expression, and inhibited NF-κB in PBC mice. These findings suggested EDNRB loss or inhibition weakened liver injury and pro-inflammatory responses by down-regulating GRK2 and inhibiting the NF-κB pathway in PBC mice.

Discovery of a Series of 5-Amide-1H-pyrazole-3-carboxyl Derivatives as Potent P2Y14R Antagonists with Anti-Inflammatory Characters.

UDPG/P2Y14R signaling pathway has been considered as a potential therapeutic target for innate immune system diseases. Based on the scaffold hopping strategy, a series of pyrazole analogues were designed and synthesized as novel P2Y14R antagonists with improved physicochemical properties, together with potential anti-inflammatory activities. Additionally, we designed and synthesized a fluorescent probe based on highly selective and potent PPTN to study the affinity of synthesized compounds. The optimized compound 16 (1-(4-fluorobenzyl)-5-(4-methylbenzamido)-1H-pyrazole-3-carboxylic acid, P2Y14R IC50 = 1.93 nM) showed strong binding ability to P2Y14R, high selectivity, notably improved solubility, and more favorable pharmacokinetic profiles. Moreover, compound 16 possessed extremely low cytotoxicity and anti-inflammatory effect in vitro. In an acute peritonitis model, compound 16 could effectively reduce the levels of inflammatory factor IL-6, IL-1β, and TNF-α of mice induced by LPS. Compound 16, with potent in vitro and in vivo efficacy and favorable druggability, can be a promising candidate for further research.

Tetrandrine alleviates inflammation and neuron apoptosis in experimental traumatic brain injury by regulating the IRE1α/JNK/CHOP signal pathway.

The aim of this study was to investigate the therapeutic roles of Tetrandrine (TET) on traumatic brain injury (TBI) and the underlying mechanism. Traumatic injury model of hippocampal neurons and TBI mouse model were established to evaluate the therapeutic effects. The expression of neuron-specific enolase (NSE), Caspase 3, and Caspase 12 was detected by immunofluorescence. The expression of TNF-α, NF-κB, TRAF1, ERS markers (GADD34 and p-PERK), IRE1α, CHOP, JNK, and p-JNK were evaluated by western blot. Flow cytometry was used to determine the apoptosis of neurons. Brain injury was assessed by Garcia score, cerebral water content, and Evan blue extravasation test. Hematoxylin and eosin staining was used to determine the morphological changes of hippocampal tissue. Apoptosis was assessed by TUNEL staining. In traumatic injury model of hippocampal neurons, TET downregulated NSE, TNF-α, NF-κB, TRAF1, GADD34, p-PERK, IRE1α, CHOP, and p-JNK expression. TET reduced Caspase 3 and Caspase 12 cleavage. Apoptosis rate was inhibited by the introduction of TET. TET improved the Garcia neural score, decreased the cerebral water content and Evans blue extravasation, and reduced NSE, TNF-α, NF-κB, TRAF1, IRE1α, CHOP, and p-JNK expression in mice with TBI, which was significantly reversed by Anisomycin, a JNK selective activator. TET alleviated inflammation and neuron apoptosis in experimental TBI by regulating the IRE1α/JNK/CHOP signal pathway.

Abstract 11626: A Hepato-Cardiac FGF21 Axis Activates a Cardiac FGF21 Autocrine Mechanism That Causes Hypertrophy With Pressure Overload

FGF21 is expressed mainly in the liver and other organs, including skeletal muscle and heart. It exerts its actions by binding to its receptors (FGFR) and the co-receptor β-klotho. Despite evidence about protective action of FGF21 in different animal disease models, elevated plasma FGF21 levels have been reported in patients with heart failure. Thus, it remains unknown whether elevation in human heart failure is part of the disease’s pathophysiology or a protective response. Staining of heart sections from end-stage heart failure patients showed increased FGF21 that correlated with higher plasma FGF21 levels. To assess the relative contribution of cardiac and extracardiac FGF21 in heart failure, we generated 3 animal models with cardiomyocyte- (CM-FGF21-KO), hepatocyte- (Alb-FGF21-KO) or skeletal muscle-specific FGF21 (MLC-FGF21-KO) deletion and subjected them to transverse aortic constriction (TAC) for 3 days, 2, 4, and 8 weeks. Hepatic expression and plasma levels of FGF21 were increased 3 days post-TAC. Cardiac FGF21 expression was also increased after 2 weeks of TAC. Surprisingly, the pro-hypertrophic effect of TAC was abolished in both Alb-FGF21-KO and CM-FGF21-KO mice. Interestingly, cardiac FGF21 expression was not increased in CM-FGF21-KO mice despite the high plasma FGF21 levels. The MLC-FGF21-KO mice were not protected. Thus, cardiomyocyte FGF21 upregulation mediates cardiac hypertrophy in response to pressure overload. To investigate if cardiomyocyte FGF21 expression is controlled by FGF21 that is released from the liver, we treated human AC16 cells with recombinant FGF21, which increased endogenous FGF21 mRNA and protein, pointing to an autoregulatory FGF21-based induction of cardiac FGF21 expression. This auto-regulatory loop was potentiated by increased expression of cardiac FGFR1 in wild-type mice subjected to TAC that was attenuated in CM-FGF21-KO mice. Accordingly, we observed increased pro-hypertrophic cardiac signaling (pAMPK, pERK,Bnp, Tnfα) in wild-type mice with TAC, but not in CM-FGF21 KO mice. Conclusively, a hepato-cardiac FGF21-based signaling axis governs cardiomyocyte FGF21 expression, which accounts for cardiac hypertrophy. Inhibition of either hepatic or cardiomyocyte FGF21 is cardioprotective.

Protective effect of genkwanin against lipopolysaccharide-induced acute lung injury in mice with p38 mitogen-activated protein kinase and nuclear factor-κB pathway inhibition

• Genkwanin inhibited the lung histopathological changes in LPS-induced ALI mice. • Genkwanin inhibited lung oedema, alveolar–capillary barrier, and leukocytes infiltration dysfunction in LPS-induced ALI mice. • Genkwanin reduced the secretion of IL-1β, IL-6, and TNF-α in LPS-induced ALI mice. • Genkwanin reduced IκB degradation and p65 phosphorylation in LPS-induced ALI mice. • Genkwanin reduced p38 MAPK phosphorylation in LPS-induced ALI mice. Acute lung injury (ALI) is a life-threatening clinical syndrome. Genkwanin is a nonglycosylated flavonoid that has several biopharmacological effects, such as anti-inflammatory, antioxidative, and immunomodulator effects. This study investigated the protective effect and mechanism of genkwanin in a lipopolysaccharide (LPS)-induced mouse model. Histopathological analysis indicated that genkwanin had a potential ameliorative effect on LPS-induced ALI. Genkwanin also inhibited several pathogenic features induced by LPS, including lung oedema, alveolar-capillary barrier dysfunction, and leukocyte and neutrophil infiltration. In addition, it inhibited NF-κB pathway activation, including phosphorylation of p65 and degradation of IκB, induced by LPS. Genkwanin also inhibited the phosphorylation of the p38 MAPK pathway induced by LPS, although it did not inhibit the phosphorylation of extracellular signal-regulated kinases or c-Jun N -terminal kinases induced by LPS. In conclusion, our results indicate that genkwanin has a protective effect against LPS-induced ALI that it exerts by inhibiting NF-κB pathway activation and p38 MAPK phosphorylation.

Semen Sojae Praeparatum improves anxiety in mice by inhibiting HPA axis hyperactivity and modulating gut microbiota

• SSP increased the desire of anxious mice to explore in elevated plus maze and the open field. • SSP exerts improved anxiety effects in mice by inhibiting HPA axis hyperactivity. • SSP decreased the levels of IL-1β, IL-6 and TNF-α in anxious mice. • SSP modulated the unbalanced gut microbiota in anxious mice. Semen Sojae Praeparatum (SSP) is a functional food made from fermented black soybeans used in folk medicine to treat psychiatric diseases. This study aimed to evaluate whether SSP affects anxiety and explain the improved anxiety mechanism of SSP. We found that SSP increased the exploratory desire of anxiety anxious mice in behavioral experiments, reduced cytokine (IL-1β, IL-6, and TNF-α) release in serum, dose-dependently decreased the protein and mRNA levels of neuronal nitric oxide synthase (nNOS), corticotropin-releasing factor (CRF), and decreased corticosterone concentrations downstream of the HPA axis. Additionally, SSP ameliorated intestinal disorders in mice by reducing the Firmicutes/Bacteroidetes ratio and increasing the abundance of Bacteroidota , Muribaculaceae , and Bacillus . These results demonstrate that SSP improved anxiety effects linked to regulation of HPA axis hyperactivity and gut microbiota composition.

Knockdown of Bcl-3 alleviates psoriasis and dyslipidemia comorbidity by regulating Akt pathway.

Psoriasis is considered as an inflammatory skin disease accompanied by dyslipidemia comorbidity. B-cell leukemia-3 (Bcl-3) belongs to IκB (inhibitor of nuclear factor kappa B [NF-κB]) family, and regulates inflammatory response through associating with NF-κB. The role of Bcl-3 in psoriasis was investigated in this study. Apolipoprotein E (ApoE)-deficient mice were treated with imiquimod to induce psoriasis and dyslipidemia. Mice were injected intradermally in the back with lentiviral particles encoding Bcl-3 small hairpin RNA (shRNA). Hematoxylin and eosin were used to detect pathological characteristics. The blood lipid levels were determined by automatic biochemical analyzer, and inflammation was assessed by enzyme-linked-immunosorbent serologic assay and real-time quantitative reverse transcription polymerase chain reaction. Bcl-3 was elevated in imiquimod-induced ApoE-deficient mice. Injection with lentiviral particles encoding Bcl-3 shRNA reduced Psoriasis area and severity index (PASI) score in ApoE-deficient psoriatic mice. Knockdown of Bcl-3 also ameliorated imiquimod-induced psoriasiform skin lesions in ApoE-deficient mice. Moreover, loss of Bcl-3 enhanced expression of loricrin, an epidermal barrier protein, reduced expression of proliferating cell nuclear antigen (PCNA) and lectin-like oxidized LDL (oxLDL) receptor-1 (LOX-1) in imiquimod-induced ApoE-deficient mice. The enhanced levels of blood lipid in ApoE-deficient mice were attenuated by silencing of Bcl-3 with increase of high-density lipoprotein, and reduction of total cholesterol, triglycerides, and low-density lipoprotein cholesterol. Knockdown of Bcl-3 attenuated imiquimod-induced decrease of transforming growth factor beta (TGF-β), and increase of Interleukin (IL)-17A, IL-23, IL-6, and tumor necrosis factor-α (TNF-α) in ApoE-deficient mice. Protein expression of phospho-Akt (p-Akt) and p-GSK3β in ApoE-deficient psoriatic mice was decreased by silencing of Bcl-3. Loss of Bcl-3 exerted anti-inflammatory effect on psoriasis and dyslipidemia comorbidity through inactivation of Akt/GSK3β pathway.

CTRP6 suppresses neutrophil extracellular traps formation to ameliorate sepsis-induced lung injury through inactivation of ERK pathway

Septic lung injury is associated with excessive neutrophil activation, while neutrophil extracellular traps formation contributes to inflammatory lung injury in sepsis. C1q/tumor necrosis factor-related protein-6 (CTRP6) is a paralog of adiponectin and exerts anti- inflammatory and antioxidant properties. The role of CTRP6 in sepsis-associated inflammatory lung injury was investigated in this study. Mice were injected with lipopolysaccharides (LPS) intraperitoneally to establish the mouse sepsis model. They were first tail-vein injected with adenovirus-mediated overexpression CTRP6 (Ad-CTRP6) and then subjected to the LPS injection. Pathological changes in lungs were detected by hematoxylin and eosin staining. Inflammation cytokine levels in bronchoalveolar lavage fluid were assessed by qRT-PCR and ELISA. Flow cytometry was used to detect the number of neutrophils in bronchoalveolar lavage fluid, and immunofluorescence was performed to assess neutrophil extracellular traps. Lipopolysaccharides induced pulmonary congestion, interstitial edema, and alveolar wall thickening in the lungs, as well as upregulated lung histology score and wet/dry weight ratio. CTRP6 was reduced in lung tissues of septic mice. Injection with Ad-CTRP6 ameliorated extensive histopathological changes in LPS-induced mice and decreased lung histology score and wet/dry weight ratio. Overexpression of CTRP6 reduced the levels of TNF-α, IL-6, and IL-1β in septic mice. Injection with Ad-CTRP6 also decreased the number of neutrophils and downregulated Cit-H3 and myeloperoxidase polymers in septic mice. Protein expression of p-ERK in septic mice was reduced by overexpression of CTRP6. CTRP6 attenuated septic lung injury, exerted anti-inflammatory effect, and suppressed neutrophil extracellular traps formation against sepsis through inactivation of extracellular signal-regulated kinase signaling.

The improved effect and its mechanism of phytic acid on DSS-induced UC mice

Ulcerative colitis (UC) is an inflammatory bowel disease (IBD) with an unknown etiology that is currently difficult to treat effectively. Phytic acid, a natural compound found in high-fiber foods, such as grains, has been shown to have many pharmacological functions, such as anti-inflammatory and blood-milk barrier maintenance. Our study investigated the effects of phytic acid on UC as well as the underlying mechanisms. We found that phytic acid ameliorated weight loss, shortened colon length, increased clinical scores, and decreased the release of pro-inflammatory factors (Il-1β, Il-6, and TNF-α) in mice with DSS-induced UC. Phytic acid was also found to protect intestinal barrier integrity in mice with UC by maintaining the expression of tight junction proteins (occludin, claudin-3 and ZO-1) and mucin-2. In LPS-stimulated Caco-2 cells, phytic acid significantly inhibited the upregulation of pro-inflammatory factors and the downregulation of tight junction proteins. Further experimental results confirmed that phytic acid inhibits the activation of the AKT/NF-κB signaling pathway in both mice with UC and Caco-2 cells. To sum up, this study proved that phytic acid has a beneficial ameliorative effect on mice with DSS-induced UC, suggesting that it may be used to develop functional foods for the treatment or prevention of UC.

Study the Ability of Salmonella typhimiurm to Induce Cytokine TNFα in Mice Treated with Olive Leaves Extract and Ciprofloxacin.

TNF-α is a type of cytokine that is produced by inflammatory cells. These inflammatory cells produce cytokines such as TNF-α, IFN- γ, IL-1, etc. Ciprofloxacin is the last drug of choice to clear the infection. Ciprofloxacin, a grace inhibitor, kills bacteria by inducing chromosome fragmentation and reactive oxygen species (ROS) in the bacterial cell. This study was designed to investigate the ability of Salmonella typhimiurm to induce TNF-α in cells. 30 NMRI mice from both genders were divided randomly into 3 groups (n=10) and treated as follows: The first group was intraperitoneally (I.P.) injected with a 0.1 ml/4×108 CFU/ ml bacterial suspension of S. typhimiurm as a positive control. The second group was injected (I.P.) with S. typhimiurm 0.1 ml (4×108 CFU/ml and then injected with 30 mg/kg/0.3 ml of CiprofloxacinCiprofloxacin intramuscular (I.M.). The olive leaf alcoholic extract was given to the third group through a gavage tube for two weeks before giving I/P 0.1 ml/4×108 CFU/ml S. typhimiurm. Afterward, they were post-treated with 30 mg/kg/0.3 ml of olive leaf extract orally by gavage tube until the end of the experiment (30 days). The recorded data from group 1 showed heavy deposition of TNF-α marker in the spleen and liver, and examination showed dark brown cellular components. Also, in the second group, the spleen and liver tissue sections showed heavy deposition of TNF-α marker; other sections showed moderated deposition of TNF-α marker. In the third group, TNF marker was found in small amounts or not in tissue samples from the spleen and liver. Histopathological examination of infected 1st group liver and spleen tissue sections shows lobular hepatic necrosis with mononuclear cells. Aggregation manifests as granuloma lesions, particularly lymphocytes in the portal area around the bile duct and blood vessels, with the proliferation of macrophages known as kupffer cells and depletion of white pulp in spleen tissue when compared to the second and third groups, which demonstrated moderated lesions in infected and treated mice. The results showed that olive leaf extract reduces the infection of Salmonella typhimiurm in the pre and post-treated groups better than CiprofloxacinCiprofloxacin.

Quality control, preparation process optimizing and anti-inflammatory effects of Premna Puberula Pamp. Pectin

BackgroundPremna Puberula Pamp. Pectin (PP) was a Wudang functional food in China. It has the effect of dispelling fire, clearing heat and detoxification in folk medicine. However, little studies have been reported for their preparation, quality control, effects and toxicity. MethodsThe P. Puberula leaves were collected from different pharms and seasons. The compounds in PP were identified using UPLC-Q-TOF-MS/MS. UV-VIS spectrophotometry with phenol-sulfuric acid and sodium nitrite aluminum nitrate were conducted for analyzing the water-soluble sugars and total flavonoids, respectively. L9(34) orthogonal experimental method was used to optimize the preparation process of PP. For the pharmacological effects of PP, the swelling right hind paw of ICR mice was modeled using subcutaneous injection of carrageenan gum solution, and the local tissue inflammatory reactions of the model mice were investigated using vernier calipers and HE staining. The serum inflammatory factor expression was detected using ELISA. The acute toxicity experiments were carried out for safety assessment of PP in ICR mice. ResultsFifty-three compounds were initially identified in PP among which flavonoids were dominant (19 out of 53). The average values of water-soluble sugar content and total flavonoid content of PP were 13.366 and 4.970 mg/g, respectively. The best preparation process of PP was powder-liquid ratio 1: 20, temperature 90 °C, and stirring time 3 min. Data showed that PP reduced paw edema and decrease the serum level of IL-6, TNF-α and IL-1β in the model mice. There was no toxic effect of PP on mice at a total dose of 6000 mg/kg/24h. ConclusionIn summary, by optimizing the preparation process, PP with stable quality can be obtained. PP has anti-inflammatory effects without toxicity.

METTL3 Attenuates Inflammation in Fusarium solani-Induced Keratitis via the PI3K/AKT Signaling Pathway.

PurposeOur previous investigations revealed a significant role of methyltransferase-like 3 (METTL3)-mediated N6-methyladenosine (m6A) modification in the development of corneal inflammation in Fusarium infection, but the exact mechanism is unknown. Therefore, this research aimed to explore how METTL3 affects the inflammatory process of fungal keratitis (FK) in mice.MethodsWe established in vitro and in vivo models by inoculating mice and primary corneal stromal cells with F. solani. METTL3 expression was confirmed by real-time quantitative polymerase chain reaction, immunofluorescence, and western blotting. After that, siRNAMETTL3 and AAV-sh-METTL3 were transfected into cells and mice to explore the role of METTL3 in the PI3K/AKT signaling pathway and inflammation. PI3K, p-PI3K, AKT, and p-AKT expression was analyzed by western blotting. Viability of corneal stromal cells was measured using a Cell Counting Kit-8 (CCK-8). Additionally, we detected interleukin (IL)-6, IL-1β, and tumor necrosis factor alpha (TNF-α) levels in corneal tissues and analyzed the role of METTL3 in inflammation in FK using slit-lamp biomicroscopy and hematoxylin and eosin staining.ResultsHere, our results show that METTL3 increased in mouse FK, and the expression of p-PI3K and p-AKT decreased when METTL3 was downregulated. We also found that knockdown of METTL3 expression attenuated the inflammatory response and decreased TNF-α, IL-1β, and IL-6 expression in corneal-infected mice. Furthermore, inhibition of the PI3K/AKT pathway attenuated the inflammatory response of FK, decreased the expression of the above inflammatory factors, and enhanced the viability of corneal stromal cells.ConclusionsBased on the study results, METTL3 downregulation attenuates Fusarium-induced corneal inflammation via the PI3K/AKT signaling pathway.

Ivermectin contributes to attenuating the severity of acute lung injury in mice

Ivermectin has been proposed as a potential anti-inflammatory drug in addition to its antiparasitic activity. Here we investigated the potential role of ivermectin in the pathogenesis of acute lung injury (ALI) using the lipopolysaccharide (LPS)- or bleomycin (BLM)-induced mice models. Male C57BL/6 mice were given ivermectin orally every day for the remainder of the experiment at doses of 1 or 2 mg/kg after 24 h of LPS or BLM treatment. Ivermectin reversed severe lung injury caused by LPS or BLM challenge, including mortality, changes in diffuse ground-glass and consolidation shadows on lung CT imaging, lung histopathological scores, lung wet/dry ratio, and protein content in the bronchoalveolar lavage fluid (BALF). Furthermore, ivermectin also reduced total lung BALF inflammatory cells, infiltrating neutrophils, myeloperoxidase activity, and plasma TNF-α and IL-6 levels in mice treated with LPS or BLM. Finally, the mechanism study showed that LPS or BLM administration increased JNK, Erk1/2, and p38 MAPK phosphorylation while decreasing IκBα expression, an inhibitor of NF-κB. However, ivermectin increased IκBα expression but blocked elevated phosphorylated JNK and p38 MAPK, not Erk1/2, in both ALI mice. These findings suggested that ivermectin may alleviate ALI caused by LPS or BLM in mice, partly via lowering the inflammatory response, which is mediated at least by the inhibition of MAPK and NF-κB signaling. Collectively, ivermectin might be used to treat acute lung injury/acute respiratory distress syndrome.