Chemical constituents from the rhizome of Acorus tatarinowii Schott and their anti-inflammatory activities in LPS induced PC12 cells

Introduction: Acorus tatarinowii Schott is a traditional Chinese medicine that aromatizes dampness, opens the body and pacifies the mind, and is used medicinally as the dried rhizome of Acorus tatarinowii, family Tenaxaceae. Research has confirmed that it has a strong anti-inflammatory activity, however, its mechanism of anti-inflammatory is still unclear, and we continue to investigate its chemical composition and anti-inflammatory mechanism. Methods: The separation and purification of the EtOH extract was carried out by preparative thin layer chromatography and column chromatography. The structures of the isolated compounds were identified by NMR spectroscopy and comparing their spectral data with those previously reported in literature. CCK-8 method was used to evaluate the anti-inflammatory activity of all compounds. The MDA, NO, SOD, IL-10, IL-1β, IL-6 and TNF-α levels were analyzed using ELISA kits. TNF-α and Beclin-1 expression were examined by western blot and immunofluorescence. Results: Ten compounds, named cinnamic acid(1), sinapic acid(2), isoferulic acid(3), α-asarone(4), ferulic acid(5), arbutin (6), caffeic acid (7), 3-(3-hydroxyl phenyl)propanol(8), ethyl caffeate(9) and β-asarone(10) were isolated from the rhizome of Acorus tatarinowii Schott. Their structures were elucidated on the basis of extensive spectroscopic analyses, as well as by comparison with literature data. The anti-inflammatory effects of the all compounds were evaluated in lipopolysaccharide(LPS)-induced PC12 cell models in our study. The results of activity test showed that compounds 3 and 6 had good inhibitory effects on 5 μg·mL−1 LPS-induced PC12 cells. The IC50 of compounds 3 and 6 against PC12 cells were 27.41 ± 0.09 µmol·L−1 and 30.19 ± 0.10 µmol·L−1, respectively. Moreover, compounds 3 and 6 showed a significant protective effect on inflammation by increasing the IL-10 and SOD levels, and reducing the TNF-α, IL-6, IL-1β, MDA and NO levels. The most important point is that their anti-inflammatory effect is related to the autophagy factor Beclin-1. Conclusion: These findings imply that compounds 3 and 6 exerts anti-inflammatory effects by regulating TNF-α/Beclin-1 pathway, which provide a new basis for the anti-inflammatory activity of Acorus tatarinowii Schott

Cytoprotective mechanism of ferulic acid against high glucose-induced oxidative stress in cardiomyocytes and hepatocytes

Background: Ferulic acid (FA), a phenolic acid, is a potential therapy for diabetes mellitus. FA has been shown to protect against hepatic and myocardial injury and oxidative stress in obese rats with late-stage diabetes, but the mechanism of the antioxidative activity of FA is still unclear. Objective: The aim of this study was to elucidate whether FA can prevent damage to cardiomyocytes and hepatocytes caused by high glucose (HG)-induced oxidative stress and whether the protection effects of FA on these cells are related to the Keap1-Nrf2-ARE signaling pathways. Design: Cells were divided into four groups: a control group (cultured with normal medium), an HG group (medium containing 80 mmol/L glucose), an FA+HG group (medium containing 80 mmol/L glucose and 1, 5, or 10 µg/mL FA), and a dimethylbiguanide (DMBG)+HG group (medium containing 80 mmol/L glucose and 50 µg/mL DMBG). Results: FA treatment significantly increased cell viability and significantly decreased cell apoptosis compared with the HG-treated group. Moreover, FA down-regulated the expression of Keap1 protein and up-regulated the expression of Nrf2 protein and gene transcription of HO-1 and glutathione S-transferase (GST) in a dose-dependent manner. Conclusion: FA alleviated the HG-induced oxidative stress and decreased cell apoptosis in hepatocytes and cardiomyocytes. These effects were associated with the Keap1-Nrf2-ARE signaling pathway

Cyanidin-3-O-glucoside protects against 1,3-dichloro-2-propanol-induced reduction of progesterone by up-regulation of steroidogenic enzymes and cAMP level in Leydig cells

1,3-Dichloro-2-propanol (1,3-DCP) is a food processing contaminant and has been shown to perturb male reproductive function. Cyanidin-3-O-glucoside (C3G), an anthocyanin antioxidant, is reported to have protective effects on many organs. However, it remains unclear whether C3G protects against chemical-induced reproductive toxicity. The present study was therefore to investigate the intervention of C3G on 1,3-DCP-induced reproductive toxicity in R2C Leydig cells. Results demonstrated that C3G inhibited the 1,3-DCP-induced cytotoxicity and cell shape damage with the effective doses being ranging from 10-40 μmol/L. In addition, 1,3-DCP (2 mmol/L) exposure significantly increased the ROS level and mitochondrial membrane potential (MMP) damage ratio, leading to a decrease in progesterone production, while C3G intervention reduced the ROS level, and increased the progesterone production after 24 h treatment. Most importantly, C3G intervention could up-regulate the cyclic adenosine monophosphate (cAMP) level and protein expression of steroidogenic acute regulatory protein (StAR) and 3β-hydroxysteroid dehydrogenase (3β-HSD). It was concluded that C3G is effective in reducing 1,3-DCP-induced reproductive toxicity via activating steroidogenic enzymes and cAMP level