High Expression Of Toll-like Receptor 4 Research Articles

Prognostic and clinicopathological significance of TLR4 expression in patients with breast cancer: a meta-analysis.

The prognostic value of Toll-like receptor 4 (TLR4) in breast cancer remains to be determined. Therefore, this paper aims to conduct a meta-analysis to assess the correlation between TLR4 and clinicopathological indicators as well as survival outcomes in breast cancer. Related literature retrieved from Embase, PubMed, Cochrane Library, Web of Science, China National Knowledge Infrastructure (CNKI) and China Wanfang. The search deadline is April 12, 2023. The outcome measures employed in the study comprised hazard ratio (HR), odds ratio (OR), and 95% confidence interval (CI) as effective indices. The data analysis was conducted using Stata 17.0 software. High TLR4 expression was associated with lymph node metastasis (OR=2.077, 95%CI=1.160-3.717, P= 0.014), tumor size (≥2 cm) (OR=2.194, 95%CI= 1.398-3.445, P= 0.001), PR expression (OR = 0.700, 95% CI = 0.505-0.971, P= 0.033), and clinical stage (OR = 3.578, 95%CI= 3.578-5.817, P<0.05), but not with histological grade (95%CI= 0.976-1.735, P= 0.072), ER expression (OR = 1.125, 95% CI = 0.492-2.571,P= 0.781), and HER-2 status (OR = 1.241, 95% CI = 0.733-2.101, P = 0.422). In addition, TLR4 overexpression was an independent prognostic indicator of DFS (HR= 1.480, 95%CI= 1.028- 2.130, p= 0.035) in breast cancer patients, but not related to OS(HR=1.730, 95%CI= 0.979-3.057, P= 0.059). From our main analysis results, high TLR4 expression is associated with lymph node metastasis, larger tumor size (≥2 cm), later clinical stage, negative PR expression and shorter DFS, suggesting poor prognosis in breast cancer patients.

Prognostic value of Toll-like receptor 4 on human monocyte subsets combined with computed tomography-adapted Leaman score assessing coronary artery disease.

Upregulation of Toll-like receptor 4 (TLR-4) is associated with coronary plaque vulnerability assessed by coronary computed tomography angiography (CCTA). Computed tomography-adapted Leaman score (CT-LeSc) is an independent long-term predictor of cardiac events. The relationship between the TLR-4 expression of CD14 ++ CD16 + monocytes and future cardiac events is unknown. We investigated this relationship using CT-LeSc in patients with coronary artery disease (CAD). We analyzed 61 patients with CAD who underwent CCTA. Three monocyte subsets (CD14 ++ CD16 - , CD14 ++ CD16 + , and CD14 + CD16 + ) and the expression of TLR-4 were measured by flow cytometry. We divided the patients into two groups according to the best cutoff value of the TLR-4 expression on CD14 + CD16 + which could predict future cardiac events. CT-LeSc was significantly greater in the high TLR-4 group than the low TLR-4 group [9.61 (6.70-13.67) vs. 6.34 (4.27-9.09), P < 0.01]. The expression of TLR-4 on CD14 ++ CD16 + monocytes was significantly correlated with CT-LeSc ( R2 = 0.13, P < 0.01). The expression of TLR-4 on CD14 ++ CD16 + monocytes was significantly higher in patients who had future cardiac events than in those who did not [6.8 (4.5-9.1) % vs. 4.2 (2.4-7.6) %, P = 0.04]. High TLR-4 expression on CD14 ++ CD16 + monocytes was an independent predictor for future cardiac events ( P = 0.01). An increase in the TLR-4 expression on CD14 ++ CD16 + monocytes is related to the development of future cardiac events.

A label-free SPR biosensor for specific detection of TLR4 expression; introducing of 10-HDA as an antagonist

Toll-like receptor 4 (TLR4) is actively involved in many health-related problems, including transplantation rejection and autoimmune diseases. Therefore, it is important to identify an antagonist to inhibit the TLR4-induced immune cell activation. In our previous study, 10-hydroxy-2-decanoic acid (10-HDA) was introduced as a potential antagonist for TLR4; however, possible interaction between 10-HDA and TLR4 needed to be detected. Due to the ability of surface plasmon resonance (SPR) biosensor to confirm the specific interactions between receptors and ligands, a new configuration of SPR biosensor proposed to detect the possible interaction between 10-HDA and TLR4. Hence, 10-HDA was immobilized using the (3-aminopropyl) triethoxysilane (APTES) polymer as a crosslinking agent on the Ag-MgF2 surface. Besides, genetically modified HEK293T cells with high TLR4 expression were used to study the possible interaction between 10-HDA and TLR4. Surprisingly, the SPR angle was significantly reduced in the presence of HEK cells expressing TLR4, while HEK cells without TLR4 did not affect the SPR angle. So, the proposed SPR biosensor successfully detected the interaction betweenTLR4 and 10-HDA. The sensitivity and detection limit of the biosensor were achieved at 0.05 and 0.5 million cells expressing TLR4, respectively, with a two-fold dynamic range.

Bench to bedside - new insights into the pathogenesis of necrotizing enterocolitis.

Necrotizing enterocolitis (NEC) is the leading cause of death and disability from gastrointestinal disease in premature infants. Recent discoveries have shed light on a unifying theorem to explain the pathogenesis of NEC, suggesting that specific treatments might finally be forthcoming. A variety of experiments have highlighted how the interaction between bacterial signalling receptors on the premature intestine and an abnormal gut microbiota incites a pro-inflammatory response in the intestinal mucosa and its underlying endothelium that leads to NEC. Central amongst the bacterial signalling receptors implicated in NEC development is the lipopolysaccharide receptor Toll-like receptor 4 (TLR4), which is expressed at higher levels in the premature gut than in the full-term gut. The high prenatal intestinal expression of TLR4 reflects the role of TLR4 in the regulation of normal gut development, and supports additional studies indicating that NEC develops in response to signalling events that occur in utero. This Review provides new evidence explaining the pathogenesis of NEC, explores new findings indicating that NEC development has origins before birth, and discusses future questions and opportunities for discovery in this field.

Protein overexpression of toll-like receptor 4 and myeloid differentiation factor 88 in oral squamous cell carcinoma and clinical significance.

Oral squamous cell carcinoma (OSCC) is the most common type of malignancy of the head and neck. In the present study, the expression of Toll-like receptor 4 (TLR4) and myeloid differentiation primary response gene 88 (MyD88) was evaluated in 55 OSCC tissues and their corresponding adjacent tissues using immunohistochemistry and reverse-transcription quantitative PCR. The results indicated that TLR4 and MyD88 were overexpressed in OSCC. Furthermore, high expression of MyD88 was negatively associated with a poor degree of differentiation, recurrence and metastasis of the tumor and was positively associated with underlying disease, including hypertension, heart disease and diabetes mellitus. Furthermore, high expression of TLR4 was positively associated with a long growth time of the tumor. In conclusion, the present study evaluated the expression levels of TLR4 and MyD88 in OSCC, as well as the association between them and clinicopathological factors, to provide markers for the prognosis and treatment of OSCC. These two genes may serve as biomarkers to optimize OSCC treatment, setting a new direction for stratifying patients and developing precise and personalized treatment regimens; the TLR4/MyD88 pathway may serve as a potential therapeutic target in the future.

Toll-Like Receptor 4 as a Favorable Prognostic Marker in Bladder Cancer: A Multi-Omics Analysis.

BackgroundThe toll-like receptor 4 (TLR4) agonist, Bacille Calmette-Guérin, has exhibited gratifying effects in treating bladder cancer. The study aims to explore the expression pattern, prognostic value, and potential mechanism of TLR4 in bladder cancer.MethodsThe transcriptome file from the GSE13507 dataset in the Gene Expression Omnibus database and the promoter methylation file from the bladder cancer dataset in The Cancer Genome Atlas database were downloaded for analysis. The prognostic value of the TLRs was assessed by univariate Cox regression. Immunohistochemistry was applied to verify the expression of TLR4 in bladder cancer. The drug response is estimated through the R package “pRRophetic.” The CIBERSORT algorithm was carried out to estimate the infiltrating immune cells of samples. Gene Set Enrichment Analysis (GSEA) was performed to identify the pathways involved under varied TLR4 expression levels.ResultsTLR4 is decreased in tumor tissues compared with surrounding tumor tissues or normal tissue, which is also positively correlated to the overall survival rate (hazard ratio [HR] = 0.38) and cancer-specific survival rate (HR = 0.15) of patients with bladder cancer. Low expression of TLR4 is observed in tumors with malignant performance (high pathological grade, higher tumor stage, and progression). Patients with low TLR4 levels are more sensitive to gemcitabine rather than cisplatin. The promoter methylation level of TLR4 is positively associated with TLR4 expression (P < 0.001). The cg14629571 methylation site largely contributes to the overall methylation level. The CIBERSORT analysis shows that high TLR4 expression is associated with lower levels of plasma cells, M0 macrophages, and M1 macrophages. The GSEA results indicate that the TGF-β pathway and apoptosis are activated in high TLR4 bladder cancer, while G2M checkpoint and E2F targets pathways are enriched in low TLR4 bladder cancer.ConclusionThis research discusses the abnormal expression and prognostic value of TLR4 in bladder cancer. The TLR4 expression can effectively predict oncological outcomes and drug sensitivity of bladder cancer patients. TLR4 is also associated with infiltrating immune cell variation and cancer pathway dysregulation. The results provide a novel prognostic marker and potential drug targets for bladder cancer.

Investigation of TLR4 and pSTAT3 expression on tumor and immune cells in the peripheral blood (PB) of patients with breast cancer (BC).

e15015 Background: Toll-like receptor 4 (TLR4) and phosphorylated signal transducer and activator of transcription protein 3 (pSTAT3) hold a key role in inflammatory response and tumor immune evasion. Their role in the peripheral immune response and in tumor immune interactions in circulation needs further investigation. In the current report we evaluated the incidence of TLR4 and pSTAT3 expression on tumor and immune compartments within the peripheral blood (PB) of patients with early (eBC) and metastatic breast cancer (mBC). Methods: PB was obtained from patients with eBC (n = 99) and mBC (n = 100), prior to adjuvant and first-line therapy, respectively. Triple immunofluorescence staining for cytokeratins (CK), TLR4 and pSTAT3 was performed on peripheral blood mononuclear cell (PBMC) cytospins. Single cell-level expression of TLR4 and pSTAT3 was assessed on CK+ circulating tumor cells (CTCs) and PBMCs using the Ariol microscopy system. Results: CK+ CTCs were detected in 6.1% and 19% of patients with eBC and mBC, respectively (p = 0.006) (number of CTCs: n = 34 and n = 28, respectively). Higher TLR4 expression rates on CTCs were observed in mBC as compared to eBC (53.6% vs 7.7% of CTCs, 13% vs 3% of patients; p = 0.016). pSTAT3+ CTCs also prevailed in mBC (60.7% vs 10.3% of CTCs; 14% vs 5.1% of patients; p = 0.032). In the metastatic patients, CTC detection was associated with shorter overall survival (median OS: 24.9 vs 36.5 months; p = 0.042; Kaplan Meier), whereas the detection of TLR4+ CTCs predicted for shorter progression free survival (median PFS: 11.4 vs 12.6 months; p = 0.036). At the PBMC level, TLR4 expression was more frequent among patients with mBC as compared to eBC (34% vs 20.2%, respectively; p = 0.029), whereas pSTAT3 expression prevailed in eBC (89.9% vs 77% of patients, respectively; p = 0.014). The detection of TLR4+ PBMCs was associated with reduced disease-free survival (median DFS: not reached; p = 0.007) and shorter OS (median OS: not reached; p = 0.028) in patients with eBC. Conclusions: TLR4 and pSTAT3 expression is demonstrated on CTCs and PBMCs and significantly varies among eBC and mBC, suggesting that the inflammatory signaling through TLR4 and pSTAT3 in PB could be associated with BC progression. Our results further imply that their assessment in PB may hold significant prognostic implications in BC.

Betaine inhibits Toll-like receptor 4 responses and restores intestinal microbiota in acute liver failure mice

Previous research has revealed that the gut microbiome has a marked impact on acute liver failure (ALF). Here, we evaluated the impact of betaine on the gut microbiota composition in an ALF animal model. The potential protective effect of betaine by regulating Toll-like receptor 4 (TLR4) responses was explored as well. Both mouse and cell experiments included normal, model, and betaine groups. The rat small intestinal cell line IEC-18 was used for in vitro experiments. Betaine ameliorated the small intestine tissue and IEC-18 cell damage in the model group by reducing the high expression of TLR4 and MyD88. Furthermore, the intestinal permeability in the model group was improved by enhancing the expression of the (ZO)-1 and occludin tight junction proteins. There were 509 operational taxonomic units (OTUs) that were identified in mouse fecal samples, including 156 core microbiome taxa. Betaine significantly improved the microbial communities, depleted the gut microbiota constituents Coriobacteriaceae, Lachnospiraceae, Enterorhabdus and Coriobacteriales and markedly enriched the taxa Bacteroidaceae, Bacteroides, Parabacteroides and Prevotella in the model group. Betaine effectively improved intestinal injury in ALF by inhibiting the TLR4/MyD88 signaling pathway, improving the intestinal mucosal barrier and maintaining the gut microbiota composition.

Mechanism of Toll-like Receptor 4 on Chronic Post-thoracotomy Pain Via Caspase-1 Activation.

Post-operative chronic post-thoracotomy pain (CPTP) has been linked to restrictions in mobility and daily activities. However, its potential causes and optimal therapy have not been well characterized. Here, the purpose of this study was to investigate the role of Toll-like receptor 4 (TLR4) in CPTP rats and its underlying mechanism. Initially, rat models of CPTP were established. Then, the mechanical withdrawal threshold (MWT) was measured after intrathecal injection of TLR4 antagonist (LPS-RS), TLR4 agonist (LPS-PG), or caspase-1 inhibitor (Ac-YVAD-CMK) in CPTP rats. Levels of TNF-α, IL-6 and IL-1β in the spinal dorsal horn (SDH) were measured by ELISA. TLR4 and caspase-1 were located by immunofluorescence double staining. TLR4 and caspase-1 levels were assessed by qRT-PCR and Western blot. TLR4 and caspase-1 were up-regulated in SDH of CPTP rats. Compared with Sham and non-CPTP groups, MWT was effectively decreased while TNF-α, IL-6 and IL-1β in SDH were increased in CPTP group. Moreover, intrathecal injection of TLR4 antagonist or caspase-1 inhibitor significantly elevated MWT expression and reduced levels of TNF-α, IL-6 and IL-1β in SDH. Additionally, high expression of TLR4 promoted mechanical hyperalgesia and inflammatory response, while intrathecal injection of a mixture of caspase-1 inhibitor and TLR4 agonist reversed the alleviation of caspase-1 inhibitor on the mechanical hyperalgesia and inflammatory response. TLR4 and caspase-1 were co-located in neurons. TLR4 aggravated CPTP in rats by mediating activation of caspase-1 in SDH.

Pain May Promote Tumor Progression via Substance P-Dependent Modulation of Toll-like Receptor-4.

In a previous study, persistent pain was suggested to be a risk factor for tumor patients. However, the mechanism underlying this phenomenon is still unclear. Substance P (SP), a pain-related neuropeptide secreted by the neural system and the immune system, plays an important role in the induction and maintenance of persistent pain. In this study, in order to explore whether SP participates in the influence of pain on tumor progression, the serum samples of lung cancer and breast cancer patients were collected and tested. An elevated expression of SP was found in patients with pain. Cell pharmacological experiments revealed that SP can upregulate the expression of Toll-like receptor-4 (TLR-4) in tumor cells and increase the proliferation, migration, and invasive activity of tumor cells. As high expression of TLR-4 has the ability to enhance the biological activity of tumor cells, TLR-4 is thought to be involved in SP-induced tumor proliferation, migration, and invasion. Treatment of tumor cells with Aprepitant, a specific blocker of the NK-1 receptor, could reduce the expression of TLR-4 and reduce the proliferation, invasion, and migration activities of tumor cells; further proof of the influence of SP on TLR-4 expression depends on the NK-1 receptor located in tumor cells. Based on the results above, we proposed a possible mechanism underlying pain affecting tumor progression: The presence of pain increases the content of SP in patients' blood, and elevated SP increases the expression of tumor TLR-4 by acting on the NK-1 receptor, which ultimately affects the biological activity of the tumor.

High TLR4 expression predicts a poor prognosis after esophagectomy for advanced thoracic esophageal squamous cell carcinoma.

Poor oral health is an independent risk factor for upper aerodigestive tract cancers, including esophageal squamous cell carcinoma (ESCC). The pattern recognition receptor Toll-like receptor 4 (TLR4) recognizes lipopolysaccharide in the cell walls of Gram-negative periodontal pathogens associated with the development and progression of ESCC. It is, therefore, plausible that TLR4 plays a crucial role in the pathogenesis of ESCC. We used an ESCC tissue microarray to confirm expression of TLR4 in patients with ESCC and to determine whether TLR4 expression status correlates with the clinicopathological features of these patients or their prognosis after esophagectomy. We also tested whether the combined expression statuses of TLR4 and TLR3 better correlate with prognosis in these patients than either parameter alone. Clinical ESCC samples from all 177 patients tested showed expression of TLR4. Moreover, high TLR4 expression (3 + and 2 +) correlated with poorer 5-year overall survival after esophagectomy than lower TLR4 expression (1 +) (p = 0.0491). Patients showing high TLR4 expression tended to have a poorer prognosis whether treated with surgery alone or with surgery and adjuvant chemotherapy. Univariate and multivariate analyses showed TLR4 expression status to be an independent prognostic factor affecting 5-year overall survival. Patients exhibiting high TLR4 expression with low TLR3 expression had a much poorer prognosis than other patients (p = < 0.0001). High TLR4 expression predicts a poor prognosis in advanced thoracic ESCC patients after esophagectomy.

Role of toll-like receptor 4 antagonist Lipopolysaccharide-Rhodobacter sphaeroides on acute stress-induced voluntary ethanol preference and drinking behaviour: In vivo Swiss Albino mouse model

The present study focused on investigating the effect of toll-like receptor 4 (TLR4) antagonist Lipopolysaccharide-Rhodobacter sphaeroides(LPS-RS) on acute, stress-induced voluntary ethanol preference and drinking behaviour, neuronal components activation, and gene expression associated with stress and addictive behaviour. This study involved the exposure of restraint stress and social isolation using Swiss Albino mice. Two-bottle choice ethanol preference analysis was used in the evaluation of voluntary ethanol seeking and drinking behaviour. Several behavioural assessments were carried out to assess fear and anxiety-like behaviour, neuromuscular ability, motor coordination and locomotion. Morphological and immunoreactivity analysis and gene expression analysis were done after the completion of behavioural assessments. TLR4 antagonist LPS-RS treated stressed-mice showed a significant decrease in ethanol drinking compared with stressed mice. Behavioural results showed that stress exposure induced fear and anxiety-like behaviour; however; no significant deficit was found on motor coordination, neuromuscular ability, locomotion and exploratory behaviour among groups. Morphological analysis showed no significant change in the prefrontal cortex and hippocampus among all groups, while immunoreactivity analysis showed higher expression of c-Fos in prefrontal cortex and hippocampus, higher TLR4 expression in the prefrontal cortex and glial fibrillary acidic protein (GFAP) in hippocampus among stressed-animals. Stressed-mice also showed significant increase in TLR4, Nuclear Factor-Kappa B (NF-kB), inducible nitric oxide synthase (iNOS), dopamine receptor D2 (DRD2), cyclic adenosine monophosphate (cAMP) response element binding protein-1 (CREB-1) and opioid receptor MU-1 (OPRM-1) genes expression compared with control and LPS-RS treated stressed-mice. As a conclusion, the antagonism of TLR4 could provide therapeutic value in the treatment of stress-induced addiction.

Vascular Cognitive Impairment Caused by Cerebral Small Vessel Disease Is Associated with the TLR4 in the Hippocampus.

Cerebral small vessel disease (CSVD) can lead to leukodystrophy and cognitive impairment. The inflammatory response mediated by Toll-like receptor 4 (TLR4) is involved in the pathological process of CSVD, but the roles of TLR4 in vascular cognitive impairment (VCI) following CSVD are not clear. To explore the roles and mechanisms of TLR4 in the development of VCI. Male spontaneous hypertension rats (SHR) and Wistar Kyoto rats (WKY) were monitored for blood pressure (BP). The spatial learning and memory were assessed every 6 weeks using Morris water maze (MWM). Blood samples from femoral artery were collected and serum was isolated. Cerebral white matter damage was evaluated using a 3.0T magnetic resonance imaging (MRI) every 12 weeks. After 35 weeks, all rats were decapitated, and the expression of TLR4 in the hippocampus was determined using western blot. The number of positive cells of TLR4, active astrocyte and microglia in hippocampus were measured using immunohistochemistry and immunofluorescence. Compared with WKY, the BP of SHR was maintained at a high level. Spatial learning and memory declined. IL-1β and TNF-α levels were elevated. Cranial coronal scanning with T2-weighted MRI showed high signal intensity in corpus callosum and external capsule of SHR. Furthermore, in SHR, the expression of TLR4, GFAP, and Iba1 in the hippocampus were increased. Hypertension can cause small vascular damage and partial white matter degeneration in the brain. SHR showed cognitive impairment with increasing age. High expression of TLR4 and glial cell response in hippocampus is one of the key mechanisms of this disease.

Effect of cerebral small vessel disease on cognitive function and TLR4 expression in hippocampus

The aim of this study was to explore the differences in cognitive and neurological functions between cerebral small vessel disease (CSVD) rats and normal rats, as well as to study the Toll- like receptor 4 (TLR4) expression in the hippocampus of CSVD rats to understand its role in this brain structure. Forty male, 12-week old, specific pathogen-free (SPF), spontaneously hypertensive rats (SHRs) were used as the observation group, while 20 12-week old, SPF, Wistar male rats were used as the control group. After each measurement, 10 rats in the observation group and 5 in the control group were decapitated to collect samples for determining the expression of TLR4 in the hippocampus. At T1, difference in the latency of rats in the observation group showed no statistically significance (p = 0.836), while at T2, T3 and T4, the observation group showed longer latency than did the control group (p < 0.001). In addition, the latency of SHRs at T1 was the shortest (p < 0.05) compared to the follow-ups (T2-T4) and were followed sequentially by T2, T3, and T4 (p < 0.05). The latency was shorter than those at T1 and T2, and the shortest latency was found at T4 (p < 0.05). CSVD rats exhibited poor cognitive and neurological functions compared to normal rats and high expression of TLR4 in the hippocampus. Based on these results, we suggest that TLR4 affects the cognitive function of CSVD rats, and, thus, may be a valuable target in the treatment of CSVD.

&lt;p&gt;TLR4 expression correlated with PD-L1 expression indicates a poor prognosis in patients with peripheral T-cell lymphomas&lt;/p&gt;

Background: Toll-like receptor 4 (TLR4), a member of the pattern recognition receptors, has been reported to be involved in carcinogenesis. However, the clinical impact of TLR4 in peripheral T-cell lymphomas (PTCL) remains unclear.Methods: The current study, using immunohistochemical staining, first examined TLR4 and programmed cell death-ligand 1 (PD-L1) expression in patients with PTCL, to correlate TLR4 and PD-L1 expression with clinicopathological parameters.Results: It was found that the rates of high expression of TLR4 and PD-L1 were 41.7% and 45.8%, respectively. TLR4 expression was closely associated with PD-L1 expression. The expression of TLR4 was closely related to primary extranodal site involvement, increased Ann Arbor stage, and low hemoglobin expression, while the expression of PD-L1 was closely related to a low platelet count and multiple extranodal organ involvements (>1). High expression of either TLR4 or PD-L1 indicated a poor survival rate for patients with PTCL. Multivariate analyses further confirmed that increased expression levels of TLR4 and PD-L1 are unfavorable prognostic factors for PTCL.Conclusion: This study demonstrates that the expressions of TLR4 and PD-L1 are independent predictors of survival time for patients with PTCL. Thus, TLR4 and PD-L1 may serve as potential therapeutic targets in PTCL patients.

Methyl 3,4-dihydroxybenzoate protects against d-galN/LPS-induced acute liver injury by inhibiting inflammation and apoptosis in mice.

Aimed to investigate the effect and mechanism of methyl 3,4-dihydroxybenzoate (MDHB) on d-galactosamine/lipopolysaccharide (d-galN/LPS)-induced acute liver failure (ALF). Confirmed the hepatoprotective effect and hepatotoxicity of MDHB by histopathological examination (HE) and examination of alanine aminotransferase (ALT) and aspartate aminotransferase (AST); the expression of serum tumour necrosis factor-alpha (TNF-α), interleukin-1 beta (IL-1β) and interleukin-6 (IL-6) was detected by ELISA; transcription levels of TNF-α, IL-1β, IL-6 and Toll-like receptor 4 (TLR4) were detected by qRT-PCR; and phosphorylation levels of p38 and p65 were analysed by Western blot. Histopathological examination and examination of ALT and AST confirmed that MDHB is a low toxicity drug that can resist d-galN/LPS-induced ALF; MDHB can effectively reduce high transcription and expression of TNF-α, IL-1β, IL-6 and TLR4 in d-galN/LPS-induced ALF; and Western blot showed that MDHB could down-regulate the expression of bax, up-regulate the expression of bcl-xl and bcl-2, and inhibit the phosphorylation of p38 and p65. Methyl 3,4-dihydroxybenzoate can effectively resist d-galN/LPS-induced acute liver failure, which is related to the inhibition of inflammation and apoptosis.

Toll-like receptor 4 and its associated proteins as prognostic factors for HCC treated by post-radiotherapy surgery.

Locally advanced hepatocellular carcinoma (HCC) treated by radiotherapy (RT) may be suited for further treatment with surgery. As a critical mediator of the post-RT immune response, Toll-like receptor 4 (TLR4) and its associated proteins may serve as prognostic factors for patients with HCC treated by post-RT surgery. In the present study, a total of 20 patients with HCC treated by post-RT surgery were enrolled. Resected tumor and peritumoral liver tissues were used to construct tissue microarrays that were assessed with immunohistochemical staining for the expression levels of TLR4, tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and vascular endothelial growth factor receptor 2 (VEGFR2). The overall (OS) and disease-free (DFS) survival outcomes for each patient were assessed, and the severity of radiation-induced liver diseases (RILDs) was detected. The patients with low TLR4 or TRAIL expression exhibited significantly better OS times than those with high TLR4 (P=0.003) or TRAIL (P=0.007) expression, whereas the median DFS times for patients with low VEGFR2 or TRAIL were significantly longer than those with high VEGFR2 (P=0.003) or TRAIL (P=0.008) expression. No significant differences in OS or DFS times were identified according to the expression of TLR4, VEGFR2 or TRAIL in peritumoral liver tissue, although more severe RILDs were identified in patients with the high expression of these factors in the peritumoral liver tissue post-RT (P<0.05). Therefore, the expression levels of TLR4 and its associated proteins in HCC tumors may be suitable as prognostic factors for patients with HCC treated by post-RT surgery. The inhibition of TLR4, VEGFR2 and TRAIL expression in HCC and non-tumor liver tissue may lessen the severity of RILDs and improve survival outcomes in the future.

Effects of combined administration of Radix Angelicae Sinensis and hydrocortisone on the therapeutic action in the murine asthma model

To investigate the effects of Radix Angelicae Sinensis (RASI) and hydrocortisone combination on the murine asthma model and the mechanism. BALB/c mice were randomly divided into control group, blood stasis model group, asthma model group, HSS group, RASI group and RASI+HSS group (n=12). Ovalbumin (OVA) was used to replicate mice asthma model and hydrocortisone sodium succinate (HSS) to copy blood stasis model. Effects of RASI, HSS and their combination on hemorheology, anti-asthma (asthmatic behaviors, lung function, lung index and water content in lung tissue) were observed. and anti-asthma mechanisms The expression of relative cytokines, high-mobility group box 1 (HMGB1), toll-like receptor 4 (TLR4) and nuclear factor-κB (NF-κB) was detected by ELISA and immunohistochemistry respectively. Eight g/kg RASI, 0.05 g/kg HSS and their combination could significantly relieve asthma behavioral indicators, improve lung function, reduce lung index and water content in lung tissue, decrease the levels of TNF-α, IL-1β and IL-6 in broncho-alveolar lavage fluid (BALF), and inhibit the high expression of HMGB1, TLR4 and NF-κB in lung tissue. The improvement of lung function and the decrease in level of relative cytokines (TNF-α、IL-1βIL-6) were better in RASI+HSS group than those in RASI group and HSS group, and the inhibition of protein expression of TLR4 and NF-κB was also too. Combined administration of RASI and hydrocortisone could decrease serum thromboxane B2 (TXB2) content and blood viscosity, which were increased induced by hydrocortisone. Combined administration of RASI and hydrocortisone have obvious anti-asthma effects and one of the mechanisms is to inhibit protein synthetization of HMGB1, TLR4 and NF-κB.The combined administration of RASI and hydrocortisone has stronger improvement of lung function than that of RASI and hydrocortisone alone, and it may be related to the inhibition of TLR4 and NF-κB synthetization. The combined administration of RASI can alleviate abnormal changes of hemorheology induced by hydrocortisone in treatment of asthma.

Paeoniflorin inhibits glioblastoma growth in vivo and in vitro: a role for the Triad3A-dependent ubiquitin proteasome pathway in TLR4 degradation.

BackgroundPaeoniflorin, a polyphenolic compound derived from Radix Paeoniae Alba (Paeonia lactiflora), has exhibited anticancer activity in various human cancers, including glioblastoma. However, the mechanisms underlying the effects of this compound have not been fully elucidated. Toll-like receptor 4 (TLR4) plays an important role in the regulation of cancer cell proliferation and progression, and high TLR4 expression in glioblastoma specimens is associated with a poor prognosis. The present study aimed to investigate whether paeoniflorin suppresses glioblastoma via inhibition of TLR4 expression.MethodsCCK-8 experiments and clone formation assay were performed to detect the cell proliferation. Western blotting was used to analyze protein expression levels. Detection of Triad3A binding with TLR4 was assessed by the immunoprecipitation. Orthotopic xenograft mouse model was used to evaluate the effect of paeoniflorin in vivo. MST was used to analyze the interaction between paeoniflorin and TLR4 protein.ResultsIn our study, we found that paeoniflorin effectively inhibited glioblastoma growth and suppressed TLR4 protein levels, as well its downstream effectors both in vivo and in vitro. Moreover, when overexpressed TLR4 in glioblastoma abolished the effects of paeoniflorin on cell proliferation, migration, and invasion. Furthermore, we found that paeoniflorin decreased TLR4 protein through ubiquitination proteasome pathway (UPP)-mediated degradation in glioblastoma cells. Mechanistically, paeoniflorin promoted Triad3A to conjugate with TLR4, resulting in degradation. In addition, Triad3A-shRNA abolished paeoniflorin-enhanced UPP-mediated TLR4 degradation. Finally, we found that paeoniflorin could directly bind with TLR4 protein as assessed by MST assay.ConclusionOur study is the first to identify a novel mechanism for the antitumor activity of paeoniflorin, specifically: it decreases tumor growth by directly targeting TLR4 and modulating the TLR4/Triad3A-dependent axis, leading to TLR4 protein degradation and inhibition of glioblastoma cell progression in vitro and in vivo. Our current findings indicate that paeoniflorin is a potential glioblastoma therapeutic agent due to its Triad3A-dependent ubiquitin degradation of TLR4.

Cardiac remodeling in obesity is associated with high gene expression of TLR‐4 and TNF‐α

AimEvaluate the correlation between the gene expression of inflammatory markers and cardiac remodeling in obese animals.Methods16 male Wistar rats were assigned to receive water plus standard diet (group C, n = 8) and Ob diet with water plus 25% sucrose (Ob group, n = 8). Weekly body weight (BW) of the animals was assessed and, after 30 weeks of experiment, adiposity index (AI). Cardiac structures measured: diastolic diameter (LVDD); diameter of the aorta (AO), left atrium (LA); The MVE index (LVMI) was calculated by normalizing to body weight estimated LV mass. The LV systolic function was assessed by cardiac output; The LV diastolic function was evaluated by the following indices: peak velocity of early diastolic filling (E wave); peak velocity of late diastolic filling (A wave); ratio between the E and A waves (E/A); deceleration time of E wave (DTE); The study was supplemented by evaluation by tissue Doppler early diastolic (E′) and late (A ′) of the mitral annulus (arithmetic average travel speeds of lateral and septal walls), and the ratio the waves (E/E′ and E′/A′). Left ventricular fragment of the animals was homogenized in TRIzol® for extraction of ribonucleic acid (RNA). After wards, the RNA was submitted to reverse transcription, conversion of RNA to complementary deoxyribonucleic acid (cDNA). cDNA was used in the polymerase chain reaction (PCR) using ready‐made assays containing TaqMan MGB (FAM) probe and primer specific for tumoral necrosis factor‐ alpha (TNF‐α) and toll‐like receptor 4 (TLR‐4).ResultsThe Ob group showed significantly higher weight (C = 493.008 ± 50.775, Ob = 562.95 ± 53.69; p &lt; 0.05) and higher adiposity index (C = 4.79 ± 0.73, Ob = 8.68 ± 1.76; p &lt; 0.05). Gene expression of TRL‐4 (C = 1.00 ± 0.337, Ob = 1.584 ± 0.559; p &lt; 0.05) and TNF‐α (C = 1.0 ± 0.485, Ob = 2.897 ± 2.596; p &lt; 0.05) presented significant increase in ob group. Results of cardiac evaluation are described in Table 1 followed by the graphs of correlation.ConclusionHigh gene expression of TLR‐4 and TNF‐α in heart is correlated with cardiac remodeling in obese animals.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.