mRNA Expression Levels Of TNF-α Research Articles

Anti-Inflammatory Effect of Tacrolimus/Hydroxypropyl-β-Cyclodextrin Eye Drops in an Endotoxin-Induced Uveitis Model.

Background: Uveitis is an infrequent disease which constitutes a major cause of ocular morbidity. Correct management is essential, being corticosteroids its cornerstone. In case of contraindication to corticosteroids or treatment failure, the use of topical tacrolimus (TAC) could be an alternative which has already demonstrated safety and effectiveness in other ocular pathologies. However, TAC eye drops are not marketed, thus their elaboration must be carried out in Hospital Pharmacy Departments (HPDs). Methods: 32 Sprague-Dawley rats were divided into 4 groups of 8 rats each: (a) untreated healthy rats (Healthy); (b) untreated Endotoxin-Induced Uveitis model-rats (EIU); (c) EIU-rats treated with standard treatment of dexamethasone ophthalmic drops (DXM) and (d) EIU-rats treated with TAC-hydroxypropyl-β-cyclodextrin eye drops previously developed by our group (TAC-HPβCD). The mRNA expression levels of IL-6, IL-8, MIP-1α and TNF-α, quantitative analysis of leucocytes in aqueous humor and histological evaluation were performed. Results: TAC-HPβCD eye drops demonstrated to reduce ocular inflammation, expression of IL-6, TNF-α, MIP-1α and leukocyte infiltration in aqueous humor. Conclusions: TAC-HPβCD eye drops showed beneficial effect in EIU model in rats, positioning as an alternative for uveitis treatment in case of corticosteroids resistance or intolerance.

Preventive Effect of Flavonoid Extract from the Peel of Gonggan (Citrus reticulata Blanco Var. Gonggan) on CCl4-Induced Acute Liver Injury in Mice.

ObjectiveCitrus peel, a waste product of citrus consumption and processing, is rich in flavonoids. This study aimed to study the protective effect of flavonoid extract from the peel of gonggan (Citrus reticulata Blanco var. gonggan) on acute chemical liver injury.Materials and MethodsWe established a chemical liver injury model induced by carbon tetrachloride (CCl4) in mice. The flavonoid composition in gonggan (Citrus reticulata Blanco var. gonggan) peel was detected by HPLC. The histopathological sections of liver, related biochemical indicators in serum and liver, and related genes were examined to evaluate the protective effect of gonggan peel flavonoid extract (GPFE).ResultsThe results showed that GPFE contained narirutin, hesperidin, nobiletin, tangeretin, and 5-demethylnobiletin. After 14 days of intragastric administration of GPFE, the result showed GPFE could reduce the increase in liver index, serum alanine aminotransferase (ALT), and aspartate transaminase (AST) levels caused by CCl4. At the same time, pathological sections of liver confirmed that GPFE alleviated the damage to liver tissue. Moreover, biochemical indicator results showed that GPFE increased the activities of superoxide dismutase (SOD) and catalase (CAT) in liver tissue and reduced the content of malondialdehyde (MDA). Also, it reduced the levels of inflammation factors: tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ), interleukin (IL)-1β, and IL-6. In addition, q-PCR results showed that GPFE upregulated mRNA expression levels of nuclear factor erythroid 2-related factor 2 (Nrf2), copper/zinc superoxide dismutase (SOD1), manganese superoxide dismutase (SOD2), glutathione peroxidase (GSH-Px), γ-glutamylcysteine synthetase (γ-GCS), CAT, and downregulated IL-6 and TNF-α mRNA expression levels. The mechanism of GPFE may be related to the inhibition of oxidative stress and inflammation.ConclusionThe experiment indicates GPFE has a good protective effect on acute chemical liver injury in mice induced by CCl4 via antioxidant and anti-inflammatory pathways.

Anti-Proliferative and Pro-Apoptotic Activities of Synthesized 3,4,5 Tri-Methoxy Ciprofloxacin Chalcone Hybrid, through p53 Up-Regulation in HepG2 and MCF7 Cell Lines.

Background:Cancer is a significant health problem around the world and one of the leading causes of human death. The need for novel, selective and non-toxic anti-cancer agents is still urging. Aim of the work: to investigate the anti-proliferative and pro-apoptotic effects of the synthesized ciprofloxacin 3,4,5 tri-methoxy chalcone hybrid (CCH) on the HepG2 hepatocellular carcinoma and MCF7 breast carcinoma cell lines. Materials and Methods:HepG2 and MCF7cell lines were treated with CCH. Cell viability and cell cycle analysis were performed. Protein and mRNA expression levels of P53, COX-2 and TNF-α were analyzed by western blotting and RT-PCR respectively. Results:CCH caused concentration and time-dependent reduction in the viability of human HepG2 and MCF7 cells, pre-G1 apoptosis and cell cycle arrest at G2/M stage, significantly higher P53 and TNF-α mRNA and protein expression levels but significantly lower COX2 mRNA and protein expression levels. Conclusion:CCH showed obvious anti-proliferative and apoptosis-inducing activities in both cell lines.

In Vivo and In Vitro Cardioprotective Effect of Gossypin Against Isoproterenol-Induced Myocardial Infarction Injury.

The aim of the study was to examine the protective effects and possible mechanism of gossypin against isoproterenol (ISO)-mediated myocardial damage in vivo and H9c2 cell damage in vitro. H9c2 cells were categorized into five groups. Viability was evaluated with MTT and LDH release in H9c2 cells. Apoptotic parameter analysis was performed with cytochrome c (Cyt-c), caspase-3 (CASP-3), and BCL2/Bax mRNA expression levels. In vivo, gossypin was administered orally to mice at doses of 5, 10, and 20mg/kg for 7days. ISO groups were injected with isoproterenol (150mg/kg) subcutaneously (on 8th and 9th) for 2days. Afterward, lactate dehydrogenase (LDH), creatine kinase-MB (CK-MB) levels and Troponin-I (Tn-I) amount from their serum, oxidative stress parameters superoxide dismutase (SOD) activity, glutathione (GSH) and malondialdehyde (MDA) levels, and tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), interleukin-1β (IL-1 β), and NF-kB mRNA expression levels with inflammatory markers from heart tissue were evaluated. In addition, IL-1B, BCL-2, and cas-3 immunohistochemical staining was performed from heart tissue and TNF-a level was measured by ELISA method. Administration of Gossypin protected the cells by dose-dependent, eliminating the reduced cell viability and increased LDH release of ISO in H9c2 cells. In mice serum analyses, increased LDH, CK-MB levels, and Tn-I levels were normalized by gossypin. ISO administration in heart tissue is regulated by gossypin with increased SOD activity, GSH amount, TNF-α, IL-6, IL-1β, and NF-kB mRNA expression levels and decreased MDA amount. Overall, the present results demonstrated that gossypin has a potential cardioprotective treatment for ischemic heart disease on in vivo and in vitro.

Phyllanthus reticulatus Prevents Ethanol-Induced Gastric Ulcer via Downregulation of IL-8 and TNF-α Levels.

The current study aimed to determine the protective effect of P. reticulatus on ethanol-induced gastric ulcer. For this purpose, thirty-six Sprague-Dawley rats were divided into six groups. The first group served as normal control, while, in other five groups, absolute ethanol was used to induce gastric ulcer. Group II served as a diseased group, while groups III, IV, and V were treated with methanol extract, ethyl acetate fraction, and n-hexane fraction, respectively, in a dose of 400 mg/kg bodyweight. Group VI was given omeprazole in a dose 20 mg/kg bodyweight. The stomachs were removed, ulcer score was evaluated, and histopathological examination of gastric lumen was conducted. Total acidity and pH values were determined in gastric juice. TNF-α and IL-8 mRNA expressions levels were determined using the reverse transcription real-time PCR method. The data indicated that P. reticulatus protected against gastric ulcer, which was evident by attenuation of ulcer score. The pretreatment with P. reticulatus raised the gastric pH and improved all evaluated histopathological parameters such as ulcer score, erosion score, hemorrhage score, fibrinoid necrosis score, inflammatory infiltrate score, and edema score. P. reticulatus significantly reduced mRNA expression levels of TNF-α and IL-8. In conclusion, P. reticulatus possess antiulcer property which might be attributed to downregulation of TNF-α and IL-8 expression levels.

CMPK2 accelerates liver ischemia/reperfusion injury via the NLRP3 signaling pathway.

Cytidine monophosphate kinase 2 (CMPK2) is a mitochondrial nucleotide monophosphate kinase which is important for the substrates of mitochondrial DNA synthesis and has been reported to participate in macrophage activation and the inflammatory response. The purpose of the present research was to determine the potential role of CMPK2 in hepatic ischemia/reperfusion (I/R) injury and to elucidate the underlying molecular mechanisms. The present study investigated the role of CMPK2 in regulating the NLRP3 pathway and liver dysfunction induced by hepatic I/R both in vivo and in vitro. It was revealed that hypoxia/reoxygenation (H/R) treatment enhanced the mRNA expression levels of CMPK2, NLRP3, IL-18, IL-1β and TNF-α in RAW 264.7 cells. The protein expression levels of IL-18, IL-1β and cleaved-caspase-1 were decreased following CMPK2 knockdown. Furthermore, the inhibition of AIM2 downregulated the expression level of IL-1β, IL-18 and cleaved-caspase-1 in the CMPK2 knockdown group followed by H/R treatment, while the inhibition of NLRP3 did not. CMPK2 deficiency also decreased alanine aminotransferase and aspartate aminotransferase expression in mice serum, as well as the pathological changes in the liver. Similarly, the release of IL-18 and IL-1β in mouse serum was also restrained with the decline of CMPK2. In conclusion, the results of the present study demonstrate that CMPK2 is indispensable for NLRP3 inflammasome activation, making CMPK2 an effective target to relieve the liver from I/R injury. In addition, the function of CMPK2 is closely associated with NLRP3 inflammasome activation, instead of AIM2.

Sophora subprostrate polysaccharide regulates histone acetylation to inhibit inflammation in PCV2-infected murine splenic lymphocytes in vitro and in vivo

Porcine circovirus type 2 (PCV2) has caused large economic losses in the swine industry worldwide; therefore, research on relevant therapeutic medicines is still urgently needed. To define the relationship between histone acetylation and inflammation induced by PCV2, we investigated whether traditional Chinese medicinal polysaccharides could alleviate viral infection by regulating histone acetylation. In this study, Sophora subprostrate polysaccharide (SSP)-treated PCV2-infected murine splenic lymphocytes in vitro and murine spleen in vivo were used to explore the regulatory effects of SSP on inflammation and histone acetylation caused by PCV2. SSP at different concentrations significantly reduced the secretion levels of the proinflammatory cytokines TNF-α and IL-6, the activity of COX-2, the mRNA expression levels of TNF-α, IL-6, iNOS and COX-2 and the protein expression levels of iNOS and COX-2 but promoted the secretion and mRNA expression levels of IL-10. Furthermore, the different concentrations of SSP significantly regulated the activity of histone acetylase (HAT) and the mRNA expression of HAT1, increased the activity of histone deacetylase (HDAC) and the mRNA expression of HDAC1 and reduced the protein expression levels of Ac-H3 and Ac-H4. Overall, SSP inhibited inflammation in PCV2-infected murine splenic lymphocytes by regulating histone acetylation in vitro and in vivo, thus playing an important role in PCV2 infection.

Smilax china L. flavonoid alleviates HFHS-induced inflammation by regulating the gut-liver axis in mice

BackgroundSmilax china L., a traditional Chinese herb, has been used to treat various inflammatory disorders, particularly pelvic inflammation. The anti-inflammatory activity of the plant extract has been reported in several in vivo experimental models. However, the underlying anti-inflammatory mechanisms and the role of gut microbiota in mice on Smilax china L. flavonoid (SCF) treatment are poorly understand. PurposeTo investigate the role of SCF in providing the anti-inflammatory response and the role of gut microbiota in high-fat/high-sucrose (HFHS)-induced obese mice for 12 weeks. Study design and methodsC57BL/6J mice were randomly divided into seven groups, normal chow (NC), HFHS, Orlistat, SCE, and low-, medium-, high- doses of SCF for 12 weeks. The body weight, liver weight, serum concentrations of lipopolysaccharide (LPS), and inflammatory cytokines in mice were assessed. The gene and protein expression levels of inflammation-related markers were measured by qRT-PCR and Western blot. Finally, the composition of gut microbiota was detected by analyzing 16S rDNA gene sequences. ResultsSCF supplement reduced body weight gain, adipose tissue and liver indexes, attenuated serum levels of tumor necrosis factor (TNF)-α, interleukin (IL)-6, LPS, and increased IL-10, and adiponectin. SCF significantly reduced the mRNA expression levels of TNF-α, IL-6, and increased the expression of AMPK, PPAR-γ, and IL-10 in mice's liver and adipose tissues. In addition, the TLR4, p-IκBα, NF-κB, and p65 protein expression levels were reduced after the SCF supplement. Moreover, SCF treatment ameliorated HFHS-induced gut dysbiosis, as revealed by an increased intestinal barrier protective species (Akkermansia spp). The relative abundance of Streptococcaceae, Faecalibaculum, and endotoxin-producing Desulfovibrionaceae were significantly decreased on SCF supplements. ConclusionThe results showed that SCF effectively inhibits HFHS-induced inflammation by suppressing the LPS-producing bacteria and pro-inflammatory bacteria group. Furthermore, the abundance of gut barrier protective species Akkermansia spp was increased to alleviate inflammatory response, inhibiting the LPS-TLR4/NF-κB signaling pathway. Thus, SCF may be a promising prophylactic for diet-induced inflammatory diseases through the gut-liver axis in mice.

Paeoniflorin Protects against ANIT-Induced Cholestatic Liver Injury in Rats via the Activation of SIRT1-FXR Signaling Pathway.

Paeoniflorin (PF), a water-soluble monoterpene glycoside, is initially isolated from the dried roots of Paeonia lactiflora Pall., which has effects on ameliorating cholestasis in our previous study. However, comprehensive approaches for understanding the protective effects and mechanisms underlying cholestatic liver injury from the regulating of bile acid metabolism have not been sufficiently elucidated. This study was aimed to explore the effectiveness as well as potential mechanism of PF on alpha-naphthylisothiocyanate (ANIT)-induced cholestatic liver injury. Rats with cholestasis induced by ANIT was used to evaluate the protective effects and mechanism of PF by regulating SIRT1/FXR and NF-κB/NLRP3 signaling pathway. Rats were intragastrically administrated with ANIT to establish cholestatic liver injury model. Serum levels of ALT, AST, TBA, TBIL, ALP, γ-GT and ALB in rats were detected. The histopathology of the liver of rats was analyzed in vivo. The relative mRNA expression and protein expression levels of IL-18, IL-1β, TNF-α, HO-1, Nrf2, TLR4, NLRP3, Caspase-1, ASC, NF-κB, FXR, and SIRT1 in liver of rats were investigated. The results showed that the serum indexes and the liver histopathology were significantly improved by PF. The overexpression of IL-18, IL-1β, TNF-α, NLRP3, ASC, and Caspase-1 in liver was markedly reduced by PF. Furthermore, PF dramatically increased the mRNA and protein expressions of SIRT1, FXR, HO-1, and Nrf2, but decreased NF-κB p65 and TLR4 levels in liver of rats. Taken together, the protective effects of PF on cholestatic liver injury were possibly related to the activation of the SIRT1/FXR and inhibition of NF-κB/NLRP3 inflammasome signaling pathway. These findings might provide a potential protection for cholestatic liver injury.

Epalrestat suppresses inflammatory response in lipopolysaccharide-stimulated RAW264.7 cells.

Lipopolysaccharide (LPS) is a potent inducer of inflammatory response. Inflammation is a major risk factor for many diseases. Regulation of inflammatory mediator and pro-inflammatory cytokine levels could be a potential therapeutic approach to treat inflammatory injury. The purpose of the present study was to determine whether epalrestat (EPS), which is used for the treatment of diabetic neuropathy, suppresses inflammatory response in LPS-stimulated RAW264.7 cells. The effects of EPS at near-plasma concentration on the levels of pro-inflammatory cytokines and inflammatory mediators was examined using by MTS assay, quantitative RT-PCR analysis, and western blotting in LPS-stimulated RAW264.7 cells. EPS suppressed mRNA and protein expression levels of pro-inflammatory cytokines, including IL-1β, IL-6, and TNFα, in RAW264.7 cells stimulated with LPS. EPS also affected inflammatory mediators such as iNOS and NF-κB in LPS-stimulated RAW264.7 cells. In this study, we demonstrated for the first time that EPS suppresses inflammatory response in LPS-stimulated RAW264.7 cells. From these results, we propose that targeting the regulation of pro-inflammatory cytokine levels and inflammatory mediators by EPS is a promising therapeutic approach to treat inflammatory injury. It is expected that EPS, whose safety and pharmacokinetics have been confirmed clinically, would be useful for the treatment of inflammatory diseases.

1-deoxynojirimycin alleviates liver fibrosis induced by type 2 diabetes in mice

To investigate the effect of 1-deoxynojirimycin (DNJ) for improving diabetic liver fibrosis and explore the underlying mechanism. Mouse models of type 2 diabetes were established in 10 Kunming mice by high-fat diet feeding for 8 weeks and intraperitoneal injection of STZ, with 5 mice receiving intraperitoneal injection of citrate buffer solution with normal feeding as the control group. The mouse models were randomized into two groups (n=5) for further highfat feeding (model group) and additional treatment with 10% DNJ in drinking water (200 mg · kg-1 per day; DNJ group) for 8 weeks. The mice were monitored for changes in body weight (BW), blood glucose, serum total cholesterol (TC), triglyceride (TG) and superoxide dismutase (SOD) levels. The pathological changes in the liver tissue were observed using HE and Sirius Red staining, and the solubility of collagens in the liver tissues was determined. The expression levels of MCP-1, TNF-α, IL-1β and TGF-β1 mRNA were detected with real-time PCR, and the protein expressions of α-SMA and collagen2 (ColA2) were determined with Western blotting. In the in vitro experiment, mouse fibroblasts L929 cells were pretreated with DNJ (10 μg/ mL) or PBS for 30 min followed by culture in high-glucose medium for 24 h, and the level of ROS production was measured using dihydroethidium (DHE) staining. In the mouse model of type 2 diabetes, DNJ treatment significantly lowered serum level of glucose, TC, and TG (P < 0.05) and increased serum SOD activity (P < 0.05). DNJ obviously attenuated liver fibrosis in the diabetic mice, as shown by alleviated cross-linking of collagens and reduced contents of pepsin-solubilized collagen (PSC) and total collagen (P < 0.05). DNJ treatment also significantly reduced the overexpression of the proinflammatory cytokines and fibrosis-related cytokines induced by diabetes (P < 0.05). In L929 cells exposed to high glucose, pretreatment with DNJ significantly lowered the intensity of red fluorescence in DHE staining. DNJ can attenuate type 2 diabetes-induced liver fibrosis in mice through its hypoglycemic, anti-inflammatory and anti-oxidative effects.

Naturally Occurring HMGB1 Inhibitor, Glycyrrhizin, Modulates Chronic Seizures-Induced Memory Dysfunction in Zebrafish Model.

Glycyrrhizin (GL) is a well-known pharmacological inhibitor of high mobility group box 1 (HMGB1) and is abundantly present in the licorice root (Glycyrrhiza radix). HMGB1 protein, a key mediator of neuroinflammation, has been implicated in several neurological disorders, including epilepsy. Epilepsy is a devastating neurological disorder with no effective disease-modifying treatment strategies yet, suggesting a pressing need for exploring novel therapeutic options. In the current investigation, using a second hit pentylenetetrazol (PTZ) induced chronic seizure model in adult zebrafish, regulated mRNA expression of HMGB1 was inhibited by pretreatment with GL (25, 50, and 100 mg/kg, ip). A molecular docking study suggests that GL establishes different binding interactions with the various amino acid chains of HMGB1 and Toll-like receptor-4 (TLR4). Our finding suggests that GL pretreatment reduces/suppresses second hit PTZ induced seizure, as shown by the reduction in the seizure score. GL also regulates the second hit PTZ induced behavioral impairment and rescued second hit PTZ related memory impairment as demonstrated by an increase in the inflection ratio (IR) at the 3 h and 24 h T-maze trial. GL inhibited seizure-induced neuronal activity as demonstrated by reduced C-fos mRNA expression. GL also modulated mRNA expression of BDNF, CREB-1, and NPY. The possible mechanism underlying the anticonvulsive effect of GL could be attributed to its anti-inflammatory activity, as demonstrated by the downregulated mRNA expression level of HMGB1, TLR4, NF-kB, and TNF-α. Overall, our finding suggests that GL exerts an anticonvulsive effect and ameliorates seizure-related memory disruption plausibly through regulating of the HMGB1-TLR4-NF-kB axis.

CCAAT/enhancer binding protein δ inhibits invasion and metastasis of liver cancer by regulating M1 type macrophages polarization

Objective: To explore the regulation of macrophage polarization and its effects on liver cancer invasion, metastasis and apoptosis by CCAAT/enhancer binding protein δ (CEBPD). Methods: THP-1 stable transfected cells with knockdown CEBPD (shCEBPD) and negative control shNC were constructed by lentviral transfection technique. THP-1 transfected cells were induced into macrophages, lipopolysaccharide (LPS) and interferon γ(IFNγ) by phorbol 12-tetradecanoate 13-acetate (PMA), and then the polarized macrophages were further induced to M1 type. The quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect M1 type macrophage related interleukin 1β (IL-1β) genes, IL-6, tumor necrosis factor α (TNFα), and inducible nitric oxide synthase (iNOS) mRNA expression level. Flow cytometry was used to detect M1 macrophage-specific surface marker CD80 expression levels. M1-induced macrophages were co-cultured with liver cancer MHCC97H cells using Transwell non-contact small sized co-culture dishes. MHCC97H cells invasion and metastasis were detected by Transwell and scratch assay under co-culture conditions, and the MHCC97H cells apoptosis was detected by flow cytometry. Results: The mRNA expression levels of M1 macrophage marker genes iNOS, TNFα, IL-6 and IL-1β in THP-1 derived macrophages were decreased after CEBPD knockdown. M1 macrophage-specific surface marker CD80 expression levels were decreased (23.7% ± 2.1% and 62.5% ± 2.0%, t = 9.58, P < 0.05). THP-1 were co-cultured with MHCC97H in shCEBPD and shNC group, respectively. Compared with shNC group, the invasion [(158.0 ± 3.5) and (75.0 ± 4.5), t = 39.87, P < 0.01] and metastatic ability (54.6% ± 1.5% and 24.3% ± 1.0%, P < 0.01) of MHCC97H cells co-cultured in shCEBPD group were stronger and the apoptosis rate was reduced [(9.4% ± 1.0%) vs. (23.7% ± 1.2%), t = 12.68, P < 0.01]. Conclusion: CEBPD can inhibit the invasion and metastasis and increase the apoptosis by amplifying M1 type macrophages polarization in liver cancer cells.

Molecular docking study and antireabsorptive activity of a semi-synthetic coumarin derivative from Platymiscium floribundum in the ligature-induced periodontitis in rats: the involvement of heme oxygenase-1.

This study aimed to evaluate the anti-resorptive activity of a semi-synthetic coumarin derivative from Platymiscium floribundum, named 6,7-dimethoxy-3-nitrocoumarin. Molecular docking studies were performed to test the binding performance of the derivative against targets associated with alveolar bone loss (TNF-α, IL-1β, and catalase) and a target considered an antioxidant defense (HO-1) during periodontitis. Periodontitis was induced by placing a nylon ligature around the second molars. The rats received for 11days 6,7-dimethoxy-3-nitrocoumarin (0.01, 0.1, or 1mg/kg) or vehicle. We investigated by RT-qPCR analysis (TNF-α, IL-1β, and HO-1 mRNA expression levels) and by colorimetric assay (catalase activity) the mechanism of action mediated by 6,7-dimethoxy-3-nitrocoumarin. The in vivo toxicity of 6,7-dimethoxy-3-nitrocoumarin was evaluated. 6,7-Dimethoxy-3-nitrocoumarin (0.1 or 1mg/kg) reduced alveolar bone loss (1.05 ± 0.24), when compared to vehicle-treated group (3.05 ± 0.30). The interactions of 6,7-dimethoxy-3-nitrocoumarin and the four targets (TNF-α, IL-1β, catalase, and HO-1) showed firm bonds above 6.0kcal/mol. 6,7-dimethoxy-3-nitrocoumarin (1mg/kg) lowered mRNA expression levels of TNF-α (2.33 ± 0.56) and IL-1β (19.87 ± 2.9), while it increased both the mRNA expression levels of HO-1 (43.40 ± 1.05) and the catalase activity (46.42 ± 4.59), when compared to vehicle-treated group (46.29 ± 8.43; 37.83 ± 4.38; 1.58 ± 0.11; 8.93 ± 1.86, respectively). The animals did not show any signs of toxicity. 6,7-Dimethoxy-3-nitrocoumarin decreased inflammatory bone loss in the ligature-induced periodontitis in rats, and the activation of the HO-1 pathway may contribute, at least partially, to its protective effects by reducing TNF-α and IL-1β mRNA levels and increasing catalase activity. 6,7-Dimethoxy-3-nitrocumarin could be used as an adjunct to subgingival instrumentation during active and supportive periodontal treatment.

Effects of dietary sodium butyrate on growth performance, antioxidant capacity, intestinal histomorphology and immune response in juvenile Pengze crucian carp (Carassius auratus Pengze)

The current study was done to assess the effects of dietary sodium butyrate (SB) on growth performance, antioxidative capacities, intestinal microvilli morphology and intestinal immune response in juvenile Pengze crucian carp (Carassius auratus Pengze). Three treated diets and a control containing 0, 1,000, 2,000, 4,000 mg/kg SB (CON, SB1, SB2, SB3) were fed for juvenile Pengze crucian carp (5.49 ± 0.41 g) for 8 weeks. The results revealed that fish in SB2 and SB3 groups showed significantly higher growth performance compared to the control (P < 0.05). The body lipid was significantly decreased in SB-containing groups compared with that of the control (P < 0.05). Trypsin and lipase activities of fish fed SB2 and SB3 diets were both significantly higher than those of fish fed CON and SB1 diets (P < 0.05). However, no significant difference was found in α-amylase activity among all groups (P > 0.05). The intestinal total antioxidant capacity, catalase and superoxide dismutase activities were significantly enhanced in SB groups compared to the control (P < 0.05). The activities of brush border enzymes such as alkaline phosphatase, γ-glutamyl transpeptidase and Na+/K+-ATPase of fish fed SB2 and SB3 diets were both significantly higher than those of fish fed CON and SB1 diets (P < 0.05). However, the malondialdehyde contents of intestines in fish fed SB diets were remarkably reduced (P < 0.05). In addition, the serum total antioxidant capacity, superoxide dismutase, lysozyme activities and complement C3 content of fish fed SB2 and SB3 diets were significantly increased than those of fish in the CON (P < 0.05). The analysis of scanning electron microscopy and transmission electron microscopy demonstrated that density and length of microvilli were increased in the SB2 and SB3 groups (P < 0.05). The mRNA expression levels of pro-inflammatory gene IL-1β was significantly down-regulated in the SB2 and SB3 groups (P < 0.05). Moreover, the mRNA expression levels of anti-inflammatory gene TGFβ and IL-10 were significantly up-regulated in SB groups (P < 0.05). The mRNA expression levels of TNFα were not significantly different among all the groups (P > 0.05). Conclusively, our results showed the positive effects of the SB in Pengze crucian carp diets to improve the growth, intestinal microvilli morphology and intestinal immune response.

The Effects of Increasing Aortic Occlusion Times at the Level of the Highest Renal Artery (Zone II) in the Normovolemic Rabbit Model

The purpose of this study was to evaluate the effects of increasing zone II resuscitative endovascular balloon occlusion of the aorta (REBOA) occlusion times on physiological, end-organ and inflammatory responses in rabbits to assess the safe aortic occlusion time in a normovolemic rabbit model. The zone ll aorta was occluded with a balloon in 32 rabbits (8 animals each for 15, 30, 60, and 90 min). 8 rabbits served as a control. ELISAs were used to examine the serum levels of ALT, AST, Cr, BUN, MDA, SOD, IL-8, IL-6, and TNF-α; HE staining was used to identify the morphological changes in the kidney; RT-PCR was used to detect the mRNA levels of IL-6, IL-8, TNF-α and NF-κB in the kidney and uterus; and Western blotting was used to measure the protein expression levels of IL-6, IL-8, TNF-α and NF-κB in the kidney and uterus. Plasma concentrations of liver markers, kidney markers, inflammatory factors and oxidative stress indicators were significantly increased at the end of reperfusion in the 30 min, 60 min and 90 min groups. Damage to the kidney occurred in the 30 min, 60 min and 90 min groups. The mRNA and protein expression levels of IL-6, IL-8, TNF-α and NF-κB in the kidney and uterus were significantly increased at the end of reperfusion in the 30 min group, and as the time of occlusion extended, these levels continued to increase. Activation of systemic inflammation and ischaemia-reperfusion injury of end-organs occurred when the occlusion time reached 30 min. Therefore, 15 min should be regarded as a safe period of REBOA in zone II.

Exposure to hexafluoropropylene oxide dimer acid (HFPO-DA) disturbs the gut barrier function and gut microbiota in mice

Hexafluoropropylene oxide dimer acid (HFPO-DA) is the substitute for perfluoro octanoic acid (PFOA), and recently it has been detected in environmental water samples worldwide and has multiple toxicities. However, whether it will affect the intestines and gut microbiota remains unclear. In this study, in order to evaluate the gut toxicity of HFPO-DA in mammals, male mice were orally exposed to 0, 2, 20, 200 μg/L HFPO-DA, respectively, for 6 weeks. Our results showed that HFPO-DA exposure caused colonic inflammation which was coupled with increased TNF-α levels in serum and increased mRNA expression levels of TNF-α, p65, TLR4, MCP-1 of the colon in mice after exposure to 200 μg/L HFPO-DA. We also found that HFPO-DA exposure induced the decreased mRNA expression levels and protein levels of MUC2 and ZO-1, which means the dysfunction of gut barrier in the colon. In the ileum, we found that HFPO-DA exposure induced the increased mRNA expression levels of various inflammatory factors, but no obvious changes was found to barrier function. Additionally, HFPO-DA exposure caused the imbalance of cecal gut microbiota and changes of cecal microbiota diversity. Taken together, all these results indicate the potential gut toxicity of HFPO-DA and is perceived as a major problem of health risk that affects the inflammation, gut barrier dysfunction, and gut microbiota disturbance in mammals.

Signaling Pathways Associated with Macrophage-Activating Polysaccharide Isolated from Korea Red Ginseng

Background and Objectives: Korean red ginseng (KRG) is known as an immune-enhancing health food and has been approved by the Korea Food and Drug Administration. We analyzed the immune-enhancing activity of KRG and its polysaccharide (KRG-P) using RAW264.7 murine macrophage cells. Materials and Methods: The protein and mRNA expression levels of IL-6 and TNF-α were measured using ELISA and qRT-PCR, respectively. Nitric oxide levels were measured using the Griess reagent. The phosphorylation and total protein levels of ERK, p38, JNK, p65, and GAPDH were determined by immunoblot assay. Results: The polysaccharide (KRG-P), but not KRG, produced nitric oxide, interleukin 6 (IL-6), and tumor necrosis factor-α (TNF-α) in RAW264.7 cells. KRG-P increased nitric oxide synthase 2 (NOS2), IL-6, and TNF-α expression in RAW264.7 cells. KRG-P also increased phosphorylation of MAPKs (mitogen-activate protein kinases) including ERK, p38, JNK, and NF-κB (nuclear factor kappa-light-chain-enhancer of activated B cells) in a concentration-dependent manner in RAW264.7 cells. Conclusions: The polysaccharide KRG-P is the active component responsible for the immune-enhancing activity of Korean red ginseng and may modulate the systemic immune system in vivo.

Morus alba L. (Sangzhi) alkaloids (SZ-A) exert anti-inflammatory effects via regulation of MAPK signaling in macrophages

Ethnopharmacological relevanceMorus alba L. (Sangzhi) alkaloids (SZ-A) tablets have been approved by the China National Medical Products Administration for T2DM treatment. Our previous study (Liu et al., 2021) revealed that SZ-A protected against diabetes and inflammation in KKAy mice. However, the mechanism and components in SZ-A exerting anti-inflammatory effects are unclear. Aim of the studyInvestigate the effects and molecular mechanisms of SZ-A on inflammation, and identify anti-inflammatory active components in SZ-A. Materials and methodsThe major ingredients in SZ-A were analyzed by HPLC and sulfuric acid - anthrone spectrophotometry. The inhibitory activities of SZ-A on lipopolysaccharide (LPS)-stimulated inflammation were determined in bone marrow-derived macrophage (BMDM) and RAW264.7 cells. The cytokine levels of IL-6 and TNF-α in cell culture supernatant were measured by enzyme-linked immunosorbent assay (ELISA). Gene expression levels of IL-6 and TNF-α were detected by qRT-PCR. The levels of protein phosphorylation of p38 MAPK, ERK, and JNK were analyzed by Western blot. ResultsThe main components in SZ-A were found to be 1-deoxynojirimycin (DNJ), 1,4-dideoxy-1,4-imino-D-arabinitol (DAB), fagomine (FAG), polysaccharide (APS), and arginine (ARG). SZ-A reduced the levels of IL-6 and TNF-α secreted by LPS-induced RAW264.7 and BMDM cells. Simultaneously, the mRNA expression levels of IL-6 and TNF-α were all significantly suppressed by SZ-A in a concentration-dependent manner. Furthermore, SZ-A inhibited the phosphorylation of p38 MAPK, ERK, and JNK in BMDM and the activation of ERK and JNK signaling in RAW264.7 cells. We also observed that DNJ, DAB, FAG, and ARG markedly downregulated IL-6 and TNF-α cytokine levels, while APS did not have an obvious effect. ConclusionsSZ-A attenuates inflammation at least partly by blocking the activation of p38 MAPK, ERK, and JNK signaling pathways. DNJ, FAG, DAB, and ARG are the main constituents in SZ-A that exert anti-inflammatory effects.

Expression and significance of let-7a and tumor necrosis factor-alpha in placenta of severe preeclampsia

Objective To investigate the expression and clinical significance of let-7a and tumor necrosis factor-alpha (TNF-α) in placental tissue of patients with severe preeclampsia (SPE). Methods From January 2018 to October 2019, 64 cases of puerperal delivery at the First People's Hospital of Lianyungang City, 44 cases of SPE (SPE group), and 20 cases of healthy pregnant women (NC group) were selected. QRT-PCR and Western-blot were used to detect the expression levels of let-7a, TNF-αmRNA, and protein in the two groups of placental tissues. The correlation and the clinical significance of the two were statistically analyzed. Results The relative expression of let-7a in the SPE group was significantly lower than that in the control group (0.06 ± 0.02 versus 0.25 ± 0.04, p < .05); the expression of TNF-αmRNA in the SPE group was significantly higher than that in the control group (0.48 ± 0.04 versus 0.17 ± 0.03, p < .05); TNF-α protein expression was significantly increased in the SPE group (1.09 ± 0.12 versus 0.56 ± 0.03, p < .05); let-7a and TNF-α were significantly negatively correlated (r = −0.41, p < .05); the blood pressure and birth weight of the pregnant women in the SPE group and the control group were significantly different [systolic blood pressure (164.27 ± 4.62) mmHg versus (125.01 ± 2.23)] mmHg, diastolic blood pressure (109.24 ± 2.97) mmHg versus (75.94 ± 2.74) mmHg, neonatal birth weight (2507.02 ± 161.46) g versus (3592.12 ± 153.05) g, p < .05]; the expression of TNF-α in placental tissue of SPE group was significantly positively correlated with systolic and diastolic blood pressure (r = 0.93 and 0.89, p < .05). Conclusion let-7a may participate in the occurrence and development of SPE by negatively regulating the expression of TNF-α.