Levels Of Inflammatory Proteins Research Articles

Exploring the Action Mechanism of the Active Ingredient of Quercetin in Ligustrum lucidum on the Mouse Mastitis Model Based on Network Pharmacology and Molecular Biology Validation.

Aim The aim of this study is to explore the mechanism of action of quercetin, the main active anti-inflammatory component of Ligustrum lucidum, in the prevention and treatment of mastitis. Methods Prediction of the main active ingredients and key anti-inflammatory targets of Ligustrum lucidum using a network pharmacology platform and molecular biology validation of the results. Observation of histopathological changes in the mouse mammary gland by hematoxylin-eosin staining(H&E) method, quantitative real-time PCR(qPCR), and Western blot (WB) to detect the expression levels of relevant inflammatory factors mRNA and protein. Results A total of 7 active ingredients and 42 key targets were obtained from the network pharmacological analysis of Ligustrum lucidum, with quercetin as the main core ingredient and tumor necrosis factor(TNF), serine threonine protein kinase1(AKT1), and interleukin6(IL6) as the core targets; H&E results showed that pathological changes were reduced to different degrees in the dose group compared to the model group. The qPCR results showed that the relative expression of TNF and IL6 mRNA in the high dose group on day 3 and the high and medium dose groups on day 7 were not significantly different compared with the blank group (P > 0.05), and the difference between the dose groups on day 5 was significant (P < 0.05). WB results showed that the difference in nuclear factor kappa-B(NF-κB) protein expression in the medium and low dose groups on day 7 was significant compared with the blank group (P < 0.05), the difference in 5 and 7 days, significant differences in AKT1 protein expression between the middle and low dose groups (P < 0.05), nonsignificant differences in the TNF protein expression between the high dose groups on day 7 (P > 0.05), and significant differences in the IL6 protein expression between the middle and low dose groups on days 3 and 7 (P < 0.05). Conclusion Quercetin, the main active ingredient of Ligustrum lucidum, may act in the prevention and treatment of mastitis by inhibiting the expression of inflammatory factors in phosphoinositol 3-kinase(PI3K)-AKT and NF-κB signaling pathways and showa a significant dose-dependent effect. This study provides theoretical basis and clues for the control of mastitis in dairy cows.

An Emerging Role of Proanthocyanidins on Psoriasis: Evidence from a Psoriasis-Like Mouse Model.

Background Psoriasis is an immune-mediated, chronic inflammatory disease, and genetic, immune, oxidative stress (OS), and environmental factors are all thought to contribute to its occurrence. Proanthocyanidins (PCs) are natural flavonoids consisting of catechins and epicatechins which have anti-inflammatory and anti-OS activities. PCs have been widely used to treat various diseases, but reports regarding psoriasis are rare. Objective To investigate the therapeutic effect and potential mechanisms of action of PCs in a psoriasis-like mouse model. Methods Thirty male BALB/c hairless mice were assigned to six groups (n = 5): normal, model, low-dose PCs, medium-dose PCs, high-dose PCs, and control groups. The final five groups were dorsally exposed to 5% imiquimod (IMQ) cream once a day for 6 consecutive days, while the normal group received no intervention. Following the first day of IMQ application, mice in the PC-treated group were dosed with different amounts of PCs daily by oral gavage for six days, whereas mice in the control group received normal saline in the same way. One week later, skin lesions were evaluated by the severity of scoring system based on psoriasis area and severity index (PASI), and pathological alterations were assessed by hematoxylin-eosin (HE) staining. Indicators of inflammation or OS, such as interleukin- (IL-) 17, IL-23, phosphorylated-phosphatidylinositol 3-kinase (p-PI3K), phosphorylated-signal transducer and activator of transcription 3 (p-STAT3), superoxide dismutase (SOD), glutathione (GSH), malondialdehyde (MDA), catalase (CAT), vascular endothelial growth factor (VEGF), inducible nitric oxide synthase (iNOS), reactive oxygen species (ROS), and heme oxygenase-1 (HO-1), were determined by ELISA, RT-PCR, western blot, and immunohistochemistry (IHC) analysis. Results IMQ administration induced the formation of large dark red plaques with thickly layered scales on the dorsal skin of mice; nevertheless, the lesions were substantially alleviated by PC administration. Histopathological alterations were observed in both model and control groups with epidermal hyperkeratosis, granulosa layer thinning, acanthosis, downward extension of rete ridges, dermal papillae expansion, capillary hyperplasia, and infiltration by inflammatory cells around blood vessels. These pathological changes, however, were restored by a range of doses of PCs, high-dose PCs in particular. Different doses of PCs significantly lowered the spleen index, levels of inflammatory or oxidative proteins (IL-17, IL-23, MDA, ROS, p-PI3K, and p-STAT3), and the mRNA expression of Il-17, Il-23, Vegf, and iNos. Protein and mRNA levels of anti-OS and anti-inflammatory biomarkers, including SOD, CAT, GSH, and HO-1, greatly increased after PC treatment, especially at the highest dose. Conclusions Our findings reveal that PCs ameliorate psoriasis-like symptoms, suppressing the inflammatory response and mitigating OS damage in an IMQ-induced psoriasis-like mouse model. These effects are probably related to the inactivation of STAT3 and PI3K and activation of HO-1 signaling.

Prognostic accuracy of inflammatory markers in predicting risk of ICU admission for COVID-19: application of time-dependent receiver operating characteristic curves.

ObjectiveIntensive care unit (ICU) admission occurs at different times during hospitalization among patients with COVID-19. We aimed to evaluate the time-dependent receive operating characteristic (ROC) curve and area under the ROC curve, AUC(t), and accuracy of baseline levels of inflammatory markers C-reactive protein (CRP) and neutrophil-to-lymphocyte ratio (NLR) in predicting time to an ICU admission event in patients with severe COVID-19 infection.MethodsIn this observational study, we evaluated 724 patients with confirmed severe COVID-19 referred to Ayatollah Rohani Hospital, affiliated with Babol University of Medical Sciences, Iran.ResultsThe AUC(t) of CRP and NLR reached 0.741 (95% confidence interval [CI]: 0.661–0.820) and 0.690 (95% CI: 0.607–0.772), respectively, in the first 3 days after hospital admission. The optimal cutoff values of CRP and NLR for stratification of ICU admission outcomes in patients with severe COVID-19 were 78 mg/L and 5.13, respectively. The risk of ICU admission was significantly greater for patients with these cutoff values (CRP hazard ratio = 2.98; 95% CI: 1.58–5.62; NLR hazard ratio = 2.90; 95% CI: 1.45–5.77).ConclusionsUsing time-dependent ROC curves, CRP and NLR values at hospital admission were important predictors of ICU admission. This approach is more efficient than using standard ROC curves.

Chickpea Protein Hydrolysate Ameliorates the Impact of Feeding a High-Fat-Diet-Induced Obesity in Mice by Modulating Inflammation

ObjectivesTo evaluate the efficacy of a chickpea (Cicer arietinum) protein hydrolysate (CPH) on ameliorating markers of obesity in a mouse model of metabolic dysfunction when fed a high fat diet. MethodsCPH was produced using an Alcalase® and Flavourzyme® dual-system. Fifty-two male C57BL/6J mice (6 weeks of age) were randomized into four groups (n = 13/group) and fed daily, for a period of 16 weeks, as follows: standard diet (SD), high-fat diet (HFD), HFD + 400 mg CPH/kg body weight (BW) (400 CPH), and HFD + 800 mg CPH/kg BW (800 CPH). BW was measured weekly and blood glucose biweekly via tail bleeding. Feces were collected during week 16. Tissue samples were collected at the end of the study. Protein expression was measured in blood plasma, and metabolites relative concentrations in feces. Gene expression analyses were performed on liver, adipose, and pancreas tissues. ResultsWeight gain for the 800 CPH group was significantly lower (19%, P < 0.01) than the weight gain in the HFD group. Both, adipose and liver tissue weights were significantly lower (12% and 26%, P < 0.05, respectively) in 800 CPH than in HFD. Group 800 CPH had significantly reduced (P < 0.05) plasma protein levels of interleukin (IL)-1 α (14%), IL-4 (35%), IL-9 (32%), lymphotactin (37%), MCP-1 (48%), MIP-1 γ (40%), and TNF- α (40%) as compared to the HFD group. In liver, the 400 CPH group had upregulation (fold change [FC] >2) of genes related to lipid metabolism (Acsl4) and adipokine signaling (Adipor2) and downregulation (FC 0.3–0.5) of genes related to insulin signaling (Pik3r1, Ikbkb, Map2k1) when compared to HFD. In adipose tissue, the 800 CPH group modulated (FC > 2) genes related to inflammation (IL1B and IL18R1), and carbohydrate metabolism (Hk2) compared to the HFD group. The 400 CPH group had significant (FC > 2) upregulation of Tlr4 in pancreas tissue as compared to HFD. More than two hundred metabolites were identified in feces; branched-chain amino acids leucine and isoleucine decreased their relative concentrations in 800 CPH in comparison to HFD. ConclusionsFindings indicated that mice fed CPH had lower BW, reduced levels of inflammatory proteins in plasma, downregulated genes related to insulin signaling, and upregulated genes related to lipid metabolism, and adipokine signaling. Funding SourcesUSDA Pulses Division.

Jelly Fig (Ficus awkeotsang Makino) Exhibits Antioxidative and Anti-Inflammatory Activities by Regulating Reactive Oxygen Species Production via NFκB Signaling Pathway.

Antioxidant and anti-inflammatory activities of Ficus awkeotsang Makino extract (FAE) on Hs68 fibroblasts and BALB/c nude-mouse models are evaluated in this study. FAE was found to be non-toxic and showed high levels of DPPH, H2O2, and hydroxyl radical scavenging abilities; a ferrous chelating capacity; as well as ferric-reducing antioxidant capability. The antioxidant activity of FAE was strongly associated with polyphenolic content (flavonoids at 10.3 mg QE g−1 and total phenol at 107.6 mg GAE g−1). The anti-inflammatory activity of FAE and the underlying molecular mechanisms were also investigated. The a* value of the mouse dorsal skin after treatment with FAE at 1.5 mg/mL in addition to chronic UVB exposure was found to decrease by 19.2% during a ten-week period. The anti-inflammatory effect of FAE was evidenced by the decreased accumulation of inflammatory cells and skin thickness. Expression levels of UVB-induced inflammatory proteins, including ROS, NF-κB, iNOS, COX-2, and IL-6, were significantly reduced upon FAE treatment in vitro and in vivo. Collectively, our results suggest that the inhibition of ROS and UVB-induced activation of the NF-κB downstream signaling pathway by FAE, indicating considerable potential as a versatile adjuvant against free radical damage in pharmaceutical applications.

Effect of Humanin G (HNG) on inflammation in age-related macular degeneration (AMD).

Inflammation plays a crucial role in the etiology and pathogenesis of AMD (Age-related Macular Degeneration). Humanin G (HNG) is a Mitochondrial Derived Peptide (MDP) that is cytoprotective in AMD and can protect against mitochondrial and cellular stress induced by damaged AMD mitochondria. The goal of this study was to test our hypothesis that inflammation-associated marker protein levels are increased in AMD and treatment with HNG leads to reduction in their protein levels. Humanin protein levels were measured in the plasma of AMD patients and normal subjects using ELISA assay. Humanin G was added to AMD and normal (control) cybrids which had identical nuclei from mitochondria-deficient ARPE-19 cells but differed in mitochondrial DNA (mtDNA) content derived from clinically characterized AMD patients and normal (control) subjects. Cell lysates were extracted from untreated and HNG-treated AMD and normal cybrids, and the Luminex XMAP multiplex assay was used to measure the levels of inflammatory proteins. AMD plasma showed reduced Humanin protein levels, but higher protein levels of inflammation markers compared to control plasma samples. In AMD RPE cybrid cells, Humanin G reduced the CD62E/ E-Selectin, CD62P/ P-Selectin, ICAM-1, TNF-α, MIP-1α, IFN–γ, IL-1β, IL-13, and IL-17A protein levels, thereby suggesting that Humanin G may rescue from mtDNA-mediated inflammation in AMD cybrids. In conclusion, we present novel findings that: A) show reduced Humanin protein levels in AMD plasma vs. normal plasma; B) suggest the role of inflammatory markers in AMD pathogenesis, and C) highlight the positive effects of Humanin G in reducing inflammation in AMD.

Effects of dezocine combined with dexmedetomidine on adverse reactions and inflammatory factors in patients undergoing HIPEC after intestinal surgery and its protective effect on the heart in the perioperative period.

The aim of this study was to explore the effects of dezocine combined with dexmedetomidine on adverse reactions and inflammatory factors in patients undergoing hyperthermic intraperitoneal chemotherapy (HIPEC) after intestinal surgery and its protective effect on the heart in the perioperative period. A total of 80 patients treated with HIPEC after intestinal surgery in our hospital from September 2018 to December 2019 were enrolled as research subjects. All patients were evenly divided into two groups using a random number table. As to analgesia and sedation during treatment, dezocine was injected intramuscularly at 30 min before treatment in the control group. Meanwhile, dezocine combined with dexmedetomidine was given in the same way in the observation group. Adverse reactions and changes in numeric rating scale (NRS) pain score during intervention were compared between the two groups. The changes in the levels of inflammatory and myocardial injury-related factors, and vascular endothelial function and regeneration ability among cardiovascular indicators at 12 h after intervention were compared as well. Additionally, the correlations of left ventricular mass index (LVMI) with the changes in the levels of inflammatory factor high-sensitivity C-reactive protein (hs-CRP), myocardial injury-related factor lactic dehydrogenase (LDH), vascular endothelial function indicator endothelin-1 (ET-1) and cardiovascular regeneration ability index vascular endothelial growth factor (VEGF) were analyzed. Compared with control group, the total prevalence rate of severe pain, respiratory depression, nausea and vomiting, diarrhea, and muscle rigidity during intervention was significantly reduced in the observation group (p<0.05). NRS pain score at 1, 4, 8 and 12 h after intervention decreased remarkably in the observation group compared with the control group (p<0.05). Meanwhile, the levels of inflammatory factors tumor necrosis factor-α (TNF-α) and hs-CRP, and myocardial injury-related factors LDH and creatine kinase MB (CKMB) as well as ET-1 at 12 h after intervention declined remarkably in observation group compared with control group (p<0.05). However, the levels of nitric oxide (NO), VEGF and basic fibroblast growth factor (bFGF) rose significantly in the observation group (p<0.05). Besides, LVMI was positively correlated with hs-CRP and LDH, whereas was negatively associated with ET-1 and VEGF (p<0.05). In HIPEC, dezocine combined with dexmedetomidine used for sedation and analgesia is able to effectively reduce adverse reactions and relieve inflammatory responses in vivo, exerting a cardio-protective effect.

Beneficial Effects of LOX‐1 Inhibition on Brain Endothelial Proinflammatory Mediators and Barrier Proteins Following OxLDL Plus Acute Ischemic Injury

Acute ischemic stroke (AIS) triggers endothelial activation and induces cerebrovascular inflammation which can result in vascular integrity loss leading to worsened stroke outcome. A clinically correlated risk factor for stroke shown to mediate vascular injury is elevated levels of oxidized low‐density lipoprotein (OxLDL). Via the lectin‐like OxLDL receptor 1 (LOX‐1), OxLDL contributes to mechanisms associated with vascular dysfunction and inflammation. The impact of OxLDL/LOX‐1 on cerebrovascular endothelial dysfunction and inflammation in the setting of AIS remains to be elucidated. The aim of this study is to investigate the effect of OxLDL via LOX‐1 on endothelial proinflammatory mediator and integral barrier protein levels during in vitro ischemic‐like conditions. We hypothesized that acute ischemic injury would increase endothelial barrier dysfunction as well as inflammation and that OxLDL would exacerbate these responses in a LOX‐1 dependent manner. Primary male human brain microvascular endothelial cells (HBMEC) were preconditioned with human OxLDL (50μg/dL; 12h) or vehicle using a serum dose reported in AIS patients. Next cells were treated with BI‐0115 (selective LOX‐1 inhibitor; 10μM) or vehicle (&lt;0.1% DMSO) for 0.5h prior to being exposed to normoxia (21% O2) or hypoxia plus glucose deprivation (HGD; 1% O2) for 6h in the continued presence of OxLDL or vehicle. Levels of HBMEC barrier protein (ZO‐1), adhesion molecule (VCAM‐1), and inflammatory proteins (iNOS, IL‐1β) were examined using FIJI mediated immunocytochemical analysis. In terms of barrier protein levels, we observed that ZO‐1 levels were decreased following HGD. Contrary to our hypothesis, OxLDL plus HGD resulted in increased levels of ZO‐1; and moreover, we observed a greater increase in ZO‐1 levels when treated with BI‐0115 alone or BI‐0115 plus OxLDL. During normoxic conditions OxLDL increased levels of ZO‐1 and this response was LOX‐1 independent. Concomitantly, HGD increased levels of the adhesion molecule, VCAM‐1. The presence of HGD plus OxLDL or OxLDL plus BI‐0115 had no effect on VCAM‐1 expression. During normoxic conditions, OxLDL increased VCAM‐1 levels in a LOX‐1 dependent manner. We next observed that HGD increased the levels of proinflammatory mediators, iNOS and IL‐1β. OxLDL under normoxic conditions significantly increased the levels of iNOS in a LOX‐1 dependent manner but had no effect on the levels of IL‐1β. HGD plus OxLDL had no effect on iNOS levels; however, treatment with BI‐0115 alone or BI‐0115 plus OxLDL significantly attenuated HGD‐mediated increases in iNOS levels. Moreover during HGD, OxLDL increased levels of IL‐1β in a LOX‐1 dependent manner suggesting that OxLDL acting through LOX‐1 leads to an increase in a central mediator of vascular inflammation and permeability. In conclusion, the beneficial effect of LOX‐1 inhibition on cerebrovascular endothelial permeability and inflammation suggests that this receptor may be a novel, viable therapeutic target in the treatment of AIS.

Distinct airway epithelial immune responses after infection with SARS-CoV-2 compared to H1N1

Children are less likely than adults to suffer severe symptoms when infected with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), while influenza A H1N1 severity is comparable across ages except for the very young or elderly. Airway epithelial cells play a vital role in the early defence against viruses via their barrier and immune functions. We investigated viral replication and immune responses in SARS-CoV-2-infected bronchial epithelial cells from healthy paediatric (n = 6; 2.5–5.6 years old) and adult (n = 4; 47–63 years old) subjects and compared cellular responses following infection with SARS-CoV-2 or Influenza A H1N1. While infection with either virus triggered robust transcriptional interferon responses, including induction of type I (IFNB1) and type III (IFNL1) interferons, markedly lower levels of interferons and inflammatory proteins (IL-6, IL-8) were released following SARS-CoV-2 compared to H1N1 infection. Only H1N1 infection caused disruption of the epithelial layer. Interestingly, H1N1 infection resulted in sustained upregulation of SARS-CoV-2 entry factors FURIN and NRP1. We did not find any differences in the epithelial response to SARS-CoV-2 infection between paediatric and adult cells. Overall, SARS-CoV-2 had diminished potential to replicate, affect morphology and evoke immune responses in bronchial epithelial cells compared to H1N1.

Therapeutic effects of the extract of Sancao Formula, a Chinese herbal compound, on imiquimod-induced psoriasis via cysteine-rich protein 61

ObjectivePsoriasis is a common chronic inflammatory skin disease that is prone to recurrence, and the proinflammatory factor, cysteine-rich protein 61 (Cyr61), is important in its pathophysiology. Long-term clinical practice has shown that Sancao Formula (SC), a Chinese herbal compound, is effective in the treatment of psoriasis, but the precise mechanism remains unknown. In this study, we investigate the mechanism by which SC extract alleviates imiquimod (IMQ)-induced psoriasis. MethodsThe expression of Cyr61 in psoriatic lesions and normal healthy skin was detected using immunohistochemical analysis to investigate the biological role of Cyr61 in models of psoriatic inflammation. A psoriatic mouse model was established by topical application of IMQ, and the effect of topical application of SC extract was evaluated using the psoriasis area and severity index (PASI) score, hematoxylin-eosin staining, and histopathological features of the skin. Next, a HaCaT cell inflammation model was established using interferon-γ (IFN-γ), and the effect of SC extract on the mRNA and protein levels of Cyr61 and intercellular cell adhesion molecule-1 (ICAM-1) was confirmed using Western blot and quantitative real-time polymerase chain reaction analyses. ResultsImmunohistochemical staining showed that the expression of Cyr61 in psoriatic lesions was higher than that in normal skin samples (78.26% vs 41.18%, P < 0.05), and the number of Cyr61-positive cells in psoriatic lesions was also significantly higher than in normal skin (18.66 ± 2.51 vs 4.33 ± 1.52, P < 0.05). Treatment in mice with IMQ-induced psoriasis showed that SC extract could significantly improve the inflammatory phenotype, PASI score (10.875 ± 0.744 vs 3.875 ± 0.582, P < 0.05), and pathological features compared with those in IMQ model group; SC treatment was also associated with decreased levels of Cyr61 and ICAM-1. In the IFN-γ-induced inflammatory cell model, the mRNA and protein levels of Cyr61 and ICAM-1 were upregulated, while the SC extract downregulated the levels of Cyr61 and ICAM-1. ConclusionThe results provide a theoretical basis for the involvement of Cyr61 in the pathogenesis of psoriasis, and suggest that SC should be used to target Cyr61 for the prevention of psoriasis recurrence.

Protein Energy Wasting in a Cohort of Maintenance Hemodialysis Patients in Dhaka, Bangladesh.

Malnutrition is associated with high rates of mortality among patients with end stage kidney disease (ESKD). There is a paucity of data from Bangladesh, where around 35,000–40,000 people reach ESKD annually. We assessed protein-energy wasting (PEW) amongst 133 patients at a single hemodialysis setting in Dhaka. Patients were 49% male, age 50 ± 13 years, 62% were on twice-weekly hemodialysis. Anthropometric, biochemical, and laboratory evaluations revealed: BMI 24.1 ± 5.2 kg/m2, mid-arm muscle circumference (MAMC) 21.6 ± 3.6 cm, and serum albumin 3.7 ± 0.6 g/dL. Based on published criteria, 18% patients had PEW and for these patients, BMI (19.8 ± 2.4 vs. 25.2 ± 5.2 kg/m2), MAMC (19.4 ± 2.4 vs. 22.2 ± 3.8 cm), serum albumin (3.5 ± 0.7 vs. 3.8 ± 0.5 g/dL), and total cholesterol (135 ± 34 vs. 159 ± 40 mg/dL), were significantly lower as compared to non-PEW patients, while hand grip strength was similar (19.5 ± 7.6 vs. 19.7 ± 7.3 kg). Inflammatory C-reactive protein levels tended to be higher in the PEW group (20.0 ± 34.8 vs. 10.0 ± 13.9 p = 0.065). Lipoprotein analyses revealed PEW patients had significantly lower low density lipoprotein cholesterol (71 ± 29 vs. 88 ± 31 mg/dL, p < 0.05) and plasma triglyceride (132 ± 51 vs. 189 ± 103 mg/dL, p < 0.05), while high density lipoprotein cholesterol was similar. Nutritional assessments using a single 24 h recall were possible from 115 of the patients, but only 66 of these were acceptable reporters. Amongst these, while no major differences were noted between PEW and non-PEW patients, the majority of patients did not meet dietary recommendations for energy, protein, fiber, and several micronutrients (in some cases intakes were 60–90% below recommendations). Malnutrition Inflammation Scores were significantly higher in PEW patients (7.6 ± 3.1 vs. 5.3 ± 2.7 p < 0.004). No discernible differences were apparent in measured parameters between patients on twice- vs. thrice-weekly dialysis. Data from a larger cohort are needed prior to establishing patient-management guidelines for PEW in this population.

Targeting Forkhead box O1-aquaporin 5 axis mitigates neuropathic pain in a CCI rat model through inhibiting astrocytic and microglial activation

ABSTRACT Forkhead box O1 (FoxO1) is a critical molecule in modulating cell growth, differentiation and metabolism, acting as a vital transcription factor. This study explored the role of FoxO1 in chronic constriction injury (CCI)-induced neuropathic pain (NP). Microglial and astrocyte activation was achieved with lipopolysaccharide (LPS, 100 ng/mL) to establish an in-vitro NP model. Morphological alterations in LPS-induced microglia and astrocytes were assayed by light microscopy. The levels of inflammatory cytokines and proteins in microglia and astrocytes were gauged by enzyme-linked immunosorbent assay (ELISA), and Western blot (WB). The CCI-induced NP rat model was constructed for investigating the FoxO1-AQP5 axis in NP. LPS markedly expanded the expression of inflammatory factors and boosted the expression of FoxO1 and AQP5 in microglia and astrocytes. Inhibition of FoxO1 or AQP5 dramatically decreased the LPS-induced inflammation in microglia and astrocytes. In vivo, CCI exacerbated the inflammatory response and NP symptoms and substantially raised the contents of FoxO1 and AQP5 in rats’ spinal cord tissues. Intrathecal administration of the Sirt1 agonist Resveratrol abated CCI-induced activation of FoxO1 and AQP5, abrogated CCI-induced mechanical hyperalgesia and thermal hyperalgesia, depressed microglial and astrocyte activation, and declined the generation of pro-inflammatory mediators in spinal cord tissues. Mechanistically, blocking the FoxO1-AQP5 pathway inactivated the ERK and p38 MAPK pathways. Suppressing the FoxO1-AQP5 axis alleviated CCI-induced NP and inflammatory responses by modulating the ERK and p38 MAPK signaling pathways.

The Impact of Aquatic Exercise Programs on the Intima-Media thickness of the Carotid Arteries, Hemodynamic Parameters, Lipid Profile and Chemokines of Community-Dwelling Older Persons: A Randomized Controlled Trial.

Scientific evidence has shown that physical exercise is an effective way of improving several cardiovascular disease markers. However, few studies have tested its effectiveness when performed in aquatic environments. The purpose of this study was to test the impact of different aquatic exercise programs on the intima-media thickness of carotid arteries (IMT) and hemodynamic and biochemical markers of cardiovascular diseases in community-dwelling older persons. A total of 102 participants were randomly allocated into four groups: an aerobic exercise group (AerG) (n = 25, 71.44 ± 4.84 years); an aerobic interval group (IntG) (n = 28, 72.64 ± 5.22 years); a combined group (ComG) (n = 29, 71.90 ± 5.67 years); and a control group (CG) (n = 20, 73.60 ± 5.25 years). The AerG, IntG, and ComG participants took part in three different aquatic exercise programs for 28 weeks. The CG participants maintained their usual routines. All participants were evaluated for IMT, blood pressure, lipid profile, and MCP-1 and MIP-1α chemokines, pre- and post-intervention. Significant differences were found in the AerG for diastolic diameter (DD), in the IntG for peak systolic velocity (PSV), and in the ComG for DD and end-diastolic velocity (EDV). Regarding blood pressure, significant differences were found in AerG for systolic blood pressure (SBP) and diastolic blood pressure (DBP); in IntG for DBP; and in ComG for SBP, DBP, and heart rate (HR). Significant differences were found in the AerG and IntG for glucose (GLU). Lower plasma levels of monocyte chemoattractant protein-1 (MCP-1) and macrophage inflammatory protein (MIP-1α) were found in the AerG and in the ComG for MCP-1 after the intervention. Aquatic physical exercise appears to improve cardiovascular health, regardless of the type of the program adopted. Aerobic programs (combined and continuous aerobic exercises) seemed to have a more beneficial effect in reducing important cardiovascular risk markers.

183 Predicting Vasospasm and Mortality in Early Severe TBI: A Model Using Serum Cytokines and Clinical Data

INTRODUCTION: It is difficult to predict outcomes in patients with severe traumatic brain injury (sTBI) using current clinical tools. METHODS: Adult civilian patients were prospectively enrolled in the sTBI arm of the Surgical Critical Care Initiative (SC2i). Patient clinical and serum inflammatory and neuronal protein data were combined and evaluated using the machine learning methods of LASSO and CART to construct parsimonious models for predicting development of post-traumatic vasospasm and mortality. Cross validations were performed to assess the robustness of conclusions. Missing data were imputed with random forest techniques; only variables with less than 10% missingness were included. RESULTS: There were 53 patients, of whom 36 (67.9%) developed vasospasm and 10 (18.9%) died. The mean age was 39.2; 22.6% were female. There were an equal number of white (25) and black (25) patients. For vasospasm, LASSO identified Eotaxin and Marshall classification of traumatic brain injury as predictors (XV AUC = 0.77). CART identified S100B as a predictor (full data AUC = 0.75). For mortality, LASSO identified S100B and sRAGE (XV AUC = 0.97), while CART identified S100B (full data AUC = 0.912). CONCLUSION: Inflammatory and glial-specific protein levels following sTBI may have predictive value that exceeds conventional clinical variables for certain outcomes. Eotaxin, S100B and radiographic findings highly predict development of post-traumatic vasospasm. S100B and sRAGE highly predict mortality. These results warrant validation in a prospective cohort.

LKB1 alleviates high glucose‑ and high fat‑induced inflammation and the expression of GnRH and sexual precocity‑related genes, in mouse hypothalamic cells by activating the AMPK/FOXO1 signaling pathway.

Precocious puberty (PP) is a developmental disorder. Hypothalamic cells can produce gonadotropin-releasing hormone (GnRH), the final output of neuroendocrine regulation that occurs during puberty. The aim of the present study was to investigate the role of live kinase B1 (LKB1), also known as serine/threonine kinase, in the progression of PP and identify the underlying mechanisms. First, the levels of LKB1 in peripheral blood and peripheral blood mononuclear cells of children with PP were detected by reverse transcription-quantitative (RT-q) PCR or western blotting. After the GT1-7 mouse hypothalamus cell line was treated with high glucose (HG) and high fat (HF), the expression of LKB1 and GnRH was tested. LKB1 was overexpressed by transfection with a pcDNA3.1 plasmid and the levels of inflammatory factors, GnRH, PP-related factors and proteins in the AMP-activated protein kinase (AMPK)/forkhead box protein O1 (FOXO1) pathway were determined using RT-qPCR or western blot analysis. Subsequently, Compound C, an inhibitor of AMPK/FOXO1 signaling, was used to clarify whether the effects of LKB1 on PP were mediated by the regulation of this pathway. Results indicated that children with PP exhibited a lower LKB1 expression. In addition, HG and HF culture resulted in an enhanced GnRH expression and a reduced LKB1 expression in GT1-7 cells. LKB1 overexpression inhibited the contents of TNF-α, IL-6 and GnRH in in GT1-7 cells exposed to HG and HF and reduced the expression of PP-related proteins, including estrogen receptor-β, cluster of differentiation 36 and G-protein-coupled receptor. In addition, the expression of phosphorylated (p)-AMPK and p-FOXO1 was markedly downregulated following LKBI overexpression. Furthermore, compound C intervention partially diminished the inhibitory effects of LKB1-mediated upregulation on the levels of inflammation and PP-related factors. In conclusion, these results demonstrated that LKB1 alleviated HG- and HF-induced inflammation, as well as the expression of GnRH and sexual precocity-related genes, in GT1-7 cells by activating the AMPK/FOXO1 signaling pathway.

MIP-1a and MIP-1b in serum as potential markers of the severe course COVID-19

PurposeStudying the pathogenesis of COVID-19 is necessary to developing of perspective predictors of severe course of disease and unfavorable prognosis. The macrophage activation syndrome observed in severe form of COVID-19 can potentially be used as a marker of poor prognosis, which makes it relevant to measure the levels of macrophage inflammatory proteins MIP-1a and MIP-1b.Methods & MaterialsStudy included 80 patients (43 men and 37 women) aged 24 -90 years (mean = 58.3 years) with laboratory confirmed COVID-19 admitted Infectious Diseases Hospital in Moscow during April - August 2020. Patients were divided into 2 groups: group 1 included patients with a moderate form (N = 30), group 2 (N = 50) included patients with a severe form of COVID-19. Serum levels of MIP-1a and MIP-1b were assessed by ELISA.ResultsAn increase of the MIP-1a level was observed in 3 patients in group 1 (10%) and in 42 patients in group 2 (84%). At the same time, the average concentration of MIP-1a was 3.71 pg/ml and 156.79 pg/ml in groups 1 and 2, respectively (p < 0.01).MIP-1b level above baseline was detected in 11 patients in group 1 (36.7%) and in 48 patients in group 2 (96%). The mean MIP-1b concentrations were 7.53 pg/ml and 152.62 pg/ml in groups 1 and 2, respectively. Similarly with MIP-1a, the difference in mean MIP-1b concentrations between the two groups was statistically significant (p < 0.01).A statistically significant correlation between the concentrations of MIP-1a and MIP-1b was observed for whole study population, the Pearson's correlation coefficient (r) is 0.756 (p < 0.01). At the same time, there were no statistically significant differences related to gender and age. Taken together, these data suggest the potential of serum concentrations of MIP-1a and MIP-1b as markers of the disease severity.ConclusionCOVID-19 is accompanied by an increase in the level of macrophage inflammatory proteins. The severe disease in most cases was associated with significant increase in the concentrations of MIP-1a and MIP-1b in the blood serum, which makes it possible to consider these proteins as potential markers of the severe COVID-19.

\u03b1-Synuclein-containing erythrocytic extracellular vesicles: essential contributors to hyperactivation of monocytes in Parkinson\u2019s disease

BackgroundImmune system dysfunction, including higher levels of peripheral monocytes and inflammatory cytokines, is an important feature of Parkinson’s disease (PD) pathogenesis, although the mechanism underlying the process remains to be investigated. In the central nervous system, it is well-known that α-synuclein (α-syn), a key protein involved in PD, activates microglia potently, and it is also reported that α-syn exists in the peripheral system, especially in erythrocytes or red blood cells (RBC) at exceedingly high concentration. The current study focused on the possibility that RBC-derived α-syn mediates the sensitization of peripheral monocytes in PD patients.MethodsThe hyperactivation of monocytes was assessed quantitatively by measuring mRNA levels of typical inflammatory cytokines (including IL-1β, IL-6 and TNF-α) and protein levels of secreted inflammatory cytokines (including pro-inflammatory cytokines: IL-1β, IL-6, TNF-α, IL-8, IFN-γ, IL-2, and IL-12p70 and anti-inflammatory cytokines: IL-4, IL-10, and IL-13). Western blot, nanoparticle tracking analysis and electron microscopy were used to characterize RBC-derived extracellular vesicles (RBC-EVs). Inhibitors of endocytosis and leucine-rich repeat kinase 2 (LRRK2), another key protein involved in PD, were used to investigate how these two factors mediated the process of monocyte sensitization by RBC-EVs.ResultsIncreased inflammatory sensitization of monocytes was observed in PD patients and PD model mice. We found that α-syn-containing RBC-EVs isolated from PD model mice or free form oligomeric α-syn induced the inflammatory sensitization of THP-1 cells, and demonstrated that endocytosis was a requirement for this pathophysiological pathway. Furthermore, the hyperactivation of THP-1 cells induced by RBC-EVs was associated with increased LRRK2 production and kinase activity. The phenomenon of inflammatory sensitization of human monocytes and increased LRRK2 were also observed by the treatment of RBC-EVs isolated from PD patients.ConclusionsOur data provided new insight into how hyperactivation of monocytes occurs in PD patients, and identified the central role played by α-syn-containing RBC-EVs in this process.

Varenicline improves cognitive impairment in a mouse model of mPFC ischemia: The possible roles of inflammation, apoptosis, and synaptic factors

Ischemia in the medial prefrontal cortex (mPFC) causes cognitive impairment in stroke cases. This study aimed to examine the effects of varenicline as α7 and α4β2 nicotine acetylcholine receptors (nAChRs) agonist, on cognitive impairment, inflammation, apoptosis, and synaptic dysfunction in mPFC ischemia. Mice were divided to three groups of control, sham, or photothrombotic mPFC ischemia model. The control and sham groups received 2 ml/kg of normal saline for a 14-day period. As well, the animals in the ischemia groups received normal saline (2 ml/kg) or varenicline at 0.1, 1, and 3 mg/kg doses for a 14-day period. Anxiety-like behaviors were then assessed by open field (OFT) and elevated plus-maze (EPM) tests. Memory was also evaluated using Morris water maze (MWM) and novel object recognition (NOR) tests. The levels of inflammatory (IL-1β, TNF-α), apoptotic (Bax, caspase3, BCL-2), and synaptic (SYP, PSD-95, and GAP-43) proteins were examined using the western blot method. In addition, the histological evaluation was performed to assess tissue damage. The administration of Varenicline at the dose of 3 mg/kg reduced the IL-1β, TNF-α, Bax, and caspase3 levels. Moreover, it increased BCL-2, SYP, PSD-95, and GAP-43 levels at the same dose and ameliorated memory impairment and anxiety-like behaviors in mPFC ischemic mice. Varenicline improved cognitive impairment by blocking inflammation and apoptosis, improving synaptic factors, and diminishing tissue damage in the mPFC ischemic mice.

Serum proteins and electrophoretic profile in horses undergoing crotalid venom hyperimmunization.

ABSTRACTIndisputably, the use of antivenoms for the treatment of snakebite envenoming is beneficial for the victims. However, there are few studies addressing the effect of long-term hyperimmunization in inoculated horses. It is known that the injection of snake venoms and adjuvants leads to local and systemic reactions in horses, but little is known about the response of inflammatory proteins. The aim of this study was to evaluate serum proteins and the electrophoretic profile of horses undergoing crotalid venom hyperimmunization. Twenty horses were divided into two groups: an inoculated group, comprising ten horses that were already being used for production of a Crotalus sp. antivenom, and a control group, comprising ten animals that had never been used for hyperimmunization. All animals were clinically healthy and without laboratory abnormalities. Total protein and albumin concentrations were measured in serum. Serum globulins were obtained by calculation. Plasma fibrinogen estimates were determined by the heat precipitation method. Serum proteinograms were obtained using agarose gel electrophoresis. The results revealed a significant increase in the concentrations of total serum proteins, globulins, and β-globulins in the inoculated group, exceeding the reference values. There were slight increases in the α-1- and α-2-globulin subfractions in serum-producing horses, with no statistical significance. We also observed that horses used to produce hyperimmune plasma developed hypoalbuminemia, although the decrease in albumin production was not statistically significant. Our findings suggest that the continuous use of horses to produce crotalid antivenom may lead to a chronic inflammatory stimulus, with changes in plasma levels of inflammatory proteins.

Effect of Sosiho-tang on a Thioacetamide-induced Liver Fibrosis Mouse Model

Objectives: The aim of the current study was to investigate the effect of Sosiho-tang on thioacetamide (TAA)-induced liver fibrosis in mice and to elucidate its underlying mechanisms.Methods: The mice were divided into 4 groups: Normal mice (Normal), TAA-induced control mice (Control), TAA-induced and silymarin-treated (50 mg/kg) mice (Silymarin), and TAA-induced and Sosiho-tang treated (200 mg/kg) mice (SSHT). Liver fibrosis was induced via intraperitoneal injection of TAA three times a week for 8 weeks. Silymarin and Sosiho-tang were concomitantly administered for 8 weeks. Serum and liver tissues were then collected and the anti-oxidant and inflammatory protein levels in the liver tissues were evaluated using western blotting.Results: SSHT administration significantly reduced the levels of AST, ALT, ammonia, and MPO in the serum. SSHT also significantly down-regulated liver NADPH oxidase and regulated the Nrf2/Keap1 signaling pathway. SSHT treatment downregulated the liver NF-κB levels and suppressed inflammatory cytokines. SSHT treatment also decreased bile acid-related factors, such as CYP7A1 and NTCP, and fibrosis-related factors, such as α-SMA and Collagen Ⅰ.Conclusions: Taken together, these data suggest that SSHT administration suppressed the progression of liver fibrosis by activating the Nrf2/Keap1 pathway and inhibiting NF-κB.