Molecular & Cellular Toxicology最新文献

Background

The BBB plays a crucial role in the development of numerous neurological diseases and is especially vulnerable to oxidative stress. Human brain microvascular endothelial cells (HBMECs), the principal constituents of the BBB, significantly contribute to its formation and preservation. Increasing evidence indicates a potential inverse correlation between the consumption of dietary flavonoids and cardiovascular risk, which could be attributed to their antioxidative properties.

Objective

To explore the impact of four prevalent and abundant flavonoids on HBMECs within a microenvironment characterized by oxidative stress.

Results

Quercetin, apigenin, and genistein notably mitigated the adverse effects of H₂O₂-induced dysfunctions observed in various HBMEC events, including capillary network differentiation, growth, and survival. Moreover, these compounds reversed the oxidative stress provoked by H₂O₂, alongside reducing oxidative damage to lipids and DNA. Conversely, myricetin failed to reverse the H₂O₂-induced oxidative stress and did not exhibit any protective effects on HBMEC. Intriguingly, quercetin and apigenin elevated NRF2 and NQO1 levels in HBMEC, while genistein did not have the same effect.

Conclusions

Our research offers preclinical evidence indicating that certain flavonoids exhibit antioxidant effects, effectively reducing the dysfunction induced by oxidative stress in brain endothelial cells. This underscores the beneficial impact of flavonoids on the blood–brain barrier (BBB). Additionally, our findings propose potential strategies utilizing flavonoids for the treatment of neurological diseases.

Background

The kidney is one of the most vulnerable organs during the pathogenesis of sepsis. Regulating podocyte injury may be helpful for the treatment of acute kidney injury (AKI) after sepsis. Liquiritin is a flavonoid isolated from the medicinal plant Glycyrrhizae Radix et Rhizoma (Gan-cao) and might have nephroprotective properties. The study aimed to explore the functions and mechanism of liquiritin in a cell model of septic AKI.

Methods

The cell model of septic AKI was established by stimulating podocytes with lipopolysaccharide (LPS). The concentration of proinflammatory factors (TNF-α, IL-1β and IL-6) was evaluated by enzyme-linked immunosorbent assay (ELISA). Podocyte viability and apoptosis were determined by cell counting kit-8 (CCK-8) and TdT-mediated dUTP nick-end labeling (TUNEL) assays. Western blotting was performed to measure the protein levels of apoptosis-related markers, nuclear factor E2-related factor 2 (Nrf2), cytoplasmic Nrf2, and nuclear Nrf2. RT-qPCR was required to assess the mRNA levels of Nrf2 and proinflammatory cytokines.

Results

LPS treatment induced podocyte injury by suppressing cell viability and accelerating cell apoptosis, and the trend was reversed by liquiritin. Moreover, liquiritin prevented LPS-evoked high levels of proinflammatory cytokines in podocytes. LPS caused the inactivation of the Nrf2 signaling by reducing cytoplasmic Nrf2 level and increasing nuclear Nrf2 level. Liquiritin activated the Nrf2 signaling in the context of LPS by controlling Nrf2 nuclear transition. Inhibition of Nrf2 signaling using ML385 suppressed the protective effect of liquiritin on podocyte dysfunction.

Conclusion

Liquiritin mitigates LPS-induced podocyte apoptosis and inflammation by activating Nrf2 signaling.

Background

Tourette syndrome (TS) is a chronic neuropsychiatric disorder, also known as multiple tic syndrome. TS is associated with the serotonin (5-HT) system, of which 5-HT2A is a subtype. Xiaoyao San (XYS) comes from “Prescriptions of the Bureau of Taiping People’s Welfare”. It has the effect of harmonizing liver and spleen, relieving depression, nourishing blood, and invigorating spleen. However, the therapeutic mechanism of XYS for TS has not been evaluated. This study aimed to investigate the therapeutic effect of XYS on the TS mouse model and its possible mechanism.

Methods

A mouse model of TS induced by DOI (selective 5-HT (2A/2C) receptor agonist, 10 ml/kg) was established. The mice were randomly divided into normal control group, DOI group, DOI + Tiapride Hydrochloride (Tia at 25 mg /kg) group, DOI + XYS low-dose (1.44 g/kg/d), medium-dose (2.88 g/kg/d) and high-dose (5.76 g/kg/d) groups, and the mice were given the corresponding drug intragastric administration for four consecutive weeks, and the stereotypic behavior was recorded weekly. The morphological characteristics of the cells were observed by hematoxylin–eosin staining. The content of 5-HT was determined by enzyme-linked immunosorbent assay. Western blot and quantitative polymerase chain reaction were used to detect 5-HT2A.

Results

XYS could significantly improve the stereotypic behavior of TS mice, and the improvement effect increased with the increase of dose, the improvement effect of XYS high dose was similar to that of Tia. After Tia or XYS high-dose treatment, the prefrontal cortex cells and neuronal structure of TS mice gradually normal, cells arranged neatly. The prefrontal cortex cells recovered generally after XYS low-dose and medium-dose treatments. 5-HT in the serum of mice in all treatment groups was higher than that in the DOI group, and 5-HT in serum of XYS high-dose group and Tia group was the highest. 5-HT2A receptor protein and mRNA in the prefrontal cortex of mice in all treatment groups showed a downward trend, and the decrease degree was the largest in the XYS high-dose group and Tia group.

Conclusion

XYS has an improvement effect on TS in mice, and its mechanism is related to reducing 5-HT2A receptor expression, increasing 5-HT content, and enhancing 5-HT system activity. These results suggest that XYS is a therapeutic strategy for TS treatment.

Objective

The present study was aimed to analyze the serum levels of malondialdehyde (MDA) as well as superoxide dismutase (SOD) among patients with non-traumatic (NT) osteonecrosis of the femoral head (ONFH).

Methods

Patients with NT-ONFH as well as healthy individuals were selected for this study. MDA and SOD levels in the serum were measured utilizing an enzyme-linked immunosorbent assay (ELISA). Besides, the severity of ONFH was evaluated radiographically utilizing the Ficat classification system. In addition, clinical severity was assessed based on the Harris hip score (HSS)/visual analogue scale (VAS).The clinical methodology for diagnostic use of MDA and SOD with regard to NT-ONFH was determined by decision tree method through R language.

Results

A total of 124 patients with NT-ONFH and 118 healthy individuals were included. NT-ONFH patients exhibited higher MDA levels but lower SOD levels in the serum when compared to healthy controls (HCs) with statistical significance. Among the Ficat stages, stage IV had notably higher MDA levels and lower SOD levels in the serum in contrast with stage III. Additionally, stage III showed increased serum MDA levels and decreased SOD levels in contrast with stage II. MDA levels in the serum in a positive relationship with the Ficat classification, while SOD levels were negatively related to it. Moreover, MDA levels in the serum were in a positive relationship with clinical severity, whereas SOD levels were negatively associated with it. Further analysis using ROC curves indicated that elevated serum MDA levels and reduced SOD levels may serve as favorable markers for assessing Ficat stage.Serum MDA > 0.57 nmol/mL along with SOD ≤ 91.15 U/mL seems to be decent for diagnosis use of NT-ONFH.

Conclusions

Patients diagnosed with NT-ONFH displayed elevated MDA levels and reduced SOD levels in the serum, indicating the involvement of increased oxidative stress and declining antioxidant activity in the progression of this condition.

Background

Cucurbitacin-I is a natural cell-permeable triterpenoid compound isolated from Cucurbitaceae and Cruciferae. The pharmacological activity of cucurbitacin-I in human ovarian cancer has been rarely reported.

Objective

The purpose of this study was to investigate the anticancer effect of cucurbitacin-I in SKOV3 human ovarian cancer cells.

Methods

SKOV3 cells were treated with different concentrations of cucurbitacin-I. Cell viability was then measured by CCK-8 assay. Apoptosis, mitochondrial membrane potential, and reactive oxygen species changed by cucurbitacin-I treatment in SKOV3 cells were measured using a Muse Cell Analyzer. Expression levels of various proteins were detected by Western blot analysis.

Results

A decrease in cell viability was observed in SKOV3 cells with cucurbitacin-I treatment. In addition, in SKOV3 cells treated with cucurbitacin-I, a significant increase in apoptosis, increased activity of various caspases, destruction of mitochondrial membrane potential, and generation of reactive oxygen species were observed compared to the control group. At the protein level, an increase in cleaved caspases and Bax/Bcl-2 ratio was induced by cucurbitacin-I treatment in SKOV3 cells. Finally, the levels of phosphorylated human epidermal growth factor receptor 2 (HER2), phosphoinositide 3-kinase (PI3K), AKT serine/threonine kinase 1 (AKT) and forkhead box O3a (FOXO3a) proteins were significantly reduced in cucurbitacin-I treated SKOV3 cells.

Conclusion

Our data demonstrate that cucurbitacin-I induces SKOV-3 cell death by inducing apoptosis via inhibition of HER2 phosphorylation and its downstream signaling molecules including PI3K, AKT and FOXO3a. This suggests a potential therapeutic role of cucurbitacin-I against ovarian cancer.

Background

Natural herb mixture extracts have been used in traditional medicine in many countries for several years and are currently widely used as alternative medicines, even in Western countries. These extracts have long been shown to be effective in alleviating and treating various diseases owing to the various components of herbal medicines. However, it is difficult to verify the efficacy of natural herb mixture extracts in clinical trials, which limits the development of new drugs.

Objective

This study was aimed at investigating the efficacy and safety of a natural herb mixture aqueous extract comprising Polyporus umbellatus (PUAE), Polygala tenuifolia (PTAE), and Epimedium koreanum (EKAE). Each of the herbs in this natural herb mixture extract is known to be cytotoxic and have mild anti-inflammatory activity when used singly. We compared the cytotoxicity of single of PUAE, PTAE and EKAE, and their mixture in RAW264.7 cells and then evaluated their possible anti-inflammatory effects.

Results

The single extracts decreased the viability of RAW264.7 cells, whereas their mixture did not affect the viability even at the highest concentration (100 ug/mL). In addition, all the tested extracts inhibited NO production and the expression of the inflammation-related proteins (iNOS and COX-2) increased by LPS stimulation. However, the effect was remarkable in cells treated with the mixture compared with cells treated with the single extracts.

Conclusion

The natural herb mixture extract comprising PUAE, PTAE and EKAE was confirmed to have a cytotoxicity-reducing effect and a synergistic anti-inflammatory effect compared with that of the individual herbal medicines that made up the natural herb mixture extract.

Background

Pancreatic adenocarcinoma (PAAD) is a malignant tumor with a very poor prognosis and lacks effective biomarkers. Testin (TES) has an altered expression in a variety of cancers, but its expression and role in PAAD remains elusive.

Objectives

To explore the prognosis and biological role of TES in PAAD.

Methods

In this study, the expression of TES in tumor tissues and the adjacent normal pancreatic tissues of PAAD, and the relationship between TES expression and PAAD overall survival (OS), were determined using gene expression profiling interactive analysis (GEPIA), human protein atlas (HPA) and Ualcan database. The correlation between TES expression and immune infiltration was assessed using tumor immunity estimation resource (TIMER) database. Protein–protein interaction (PPI) network of TES was constructed using the Search Tool for the Retrieval of Interacting Genes (STRING) database. Tissue and blood samples from PAAD or healthy subjects were collected, and the expression of signature genes was analyzed using qRT–PCR.

Results

Our results showed that TES overexpression occurred in PAAD, and predicted worse prognosis. TES expression was positively correlated with the levels of infiltrating B cells, CD8+ T cells, macrophages, neutrophils and dendritic cells. The top six hub genes HSPB1, VASP, RAB2A, ENAH, ZPR1 and THADA that interact with TES were identified, and they all were overexpression in PAAD, but only VASP expression was negatively correlated with PAAD prognosis. In our samples, compared with the adjacent normal tissues, a higher expression of TES and VASP in PAAD tumor tissues was validated.

Conclusions

The present study suggested that TES could function as a supporter and prognostic marker of PAAD, and it might work via inducing immune infiltration.

Background

Obesity refers to a range of disorders associated with fatty liver, which occur in abnormal and accumulated excessive fat as well as impaired intestinal epithelial barrier function, contributing to host systemic inflammation and metabolic dysfunction.

Objective

In this study, we examined the effects of Oncheong-eum (OCE) on lipid metabolism and barrier protection in obese mice fed a high-fat diet of OCE. C57BL/6 J male mice were fed a normal diet (10% Kcal%fat) and a high-fat diet (45% Kcal%fat) for 14 weeks to induce obesity. Serum, hepatic, and intracellular lipid contents were assessed. Histopathological staining was used to evaluate the extent of hepatic lipid accumulation and intestinal barrier integrity. Real-time polymerase chain reaction (PCR) and Western blotting were conducted to examine the expression of factors associated with lipid metabolism and tight junction protection.

Results

The body weight in the high-fat diet group significantly increased compared with that in the normal diet group, confirming that obesity was induced. On the contrary, body weight in the OCE-treated group significantly decreased compared with that in the high-fat diet control group. Liver and adipose tissue weight in the OCE-treated group was also significantly reduced compared with that in the high-fat diet control group. The serum triglyceride, total cholesterol, and liver triglyceride concentrations of the OCE-treated group were significantly decreased compared with those of the high-fat diet control group, but the serum HDL-cholesterol concentration was significantly increased. In addition, the length of the colon increased, which had been reduced because of a high-fat diet, and the intestinal wall was protected by regulating tight junction genes.

Conclusion

OCE reduces body weight, increases the weight of organs when fed a high-fat diet, and improves the lipid composition of the serum and liver. Therefore, OCE has shown great application potential as a material for obesity-related medicines and health-functional food.

Background

Baicalin has been proven to have the potential to reduce apoptosis and diabetic cardiomyopathy (DCM). However, the mechanism behind this effect still needs to be fully understood.

Objectives

To explore the potential therapeutic properties of Baicalin in managing DCM and controlling glycemic levels.

Results

In this study, Baicalin (at doses of 20, 60, or 120 mg/kg/d) were used to treat diabetic rats. At the end of treatment, the heart function of the rats was assessed. Furthermore, their serum levels of TG, TC, and LDL were measured using the ELISA method. Cell viability was evaluated using the CCK8 assay and apoptosis was assessed using flow cytometry or TUNEL assay. Primary cardiomyocytes were infected with NRF2 siRNA and then treated with Baicalin while incubating with high glucose (25 mmol/L). Protein and mRNA variations were analyzed using Western blot and qRT-PCR, respectively. The study found that when given Baicalin, diabetic rats demonstrated improved heart function. Without treatment, the hearts of diabetic rats displayed elevated levels of apoptotic cell death and cardiomyocyte autophagy, as well as decreased expressions of NRF2, HO-1, and KEAP1. However, Baicalin was able to reverse all of these diabetes-induced biochemical changes. Treatment enhanced NRF2 nuclear transfer, reduced hyperglycemia-induced apoptosis and autophagy in primary cardiomyocytes, and improved cellular viability in in vitro experiments. It must be noted that the protective effects of Baicalin were only observed when the Nrf2 gene expression was present in primary cardiomyocytes.

Conclusion

Baicalin may reduce the effects of DCM by activating NRF2 through KEAP1 suppression and regulating autophagy activation.

Background

Glioblastoma (GBM) is the most aggressive form of glioma. However, the treatment of GBM remains largely ineffective. LTBP2 is a secreted extracellular matrix protein involved in the progression of multiple tumors. However, the role and mechanism of LTBP2 in glioma remain unknown.

Objective

To uncover the possible effects of LTBP2 on the progression of gliomas and explore its underlying mechanism.

Results

LTBP2 was highly expressed in glioma. LTBP2 knockdown inhibited the growth of glioma cells. Downregulation of LTBP2 suppressed glioma cell motility. LTBP2 knockdown restrained the PI3K/AKT/mTOR pathway. LTBP2 knockdown suppressed glioma growth and migration and tumor growth in mice through the PI3K/AKT/mTOR axis.

Conclusion

LTBP2 could serve as a potential therapeutic target for the treatment of gliomas.