Punicalagin, a hydrolysable tannin of pomegranate juice, exhibits multiple biological effects, including inhibiting production of pro-inflammatory cytokines in macrophages. Autophagy, an intracellular self-digestion process, has been recently shown to regulate inflammatory responses. In this study, we investigated the anti-inflammatory potential of punicalagin in lipopolysaccharide (LPS) induced RAW264.7 macrophages and uncovered the underlying mechanisms. Punicalagin significantly attenuated, in a concentration-dependent manner, LPS-induced release of NO and decreased pro-inflammatory cytokines TNF-α and IL-6 release at the highest concentration. We found that punicalagin inhibited NF-κB and MAPK activation in LPS-induced RAW264.7 macrophages. Western blot analysis revealed that punicalagin pre-treatment enhanced LC3II, p62 expression, and decreased Beclin1 expression in LPS-induced macrophages. MDC assays were used to determine the autophagic process and the results worked in concert with Western blot analysis. In addition, our observations indicated that LPS-induced releases of NO, TNF-α, and IL-6 were attenuated by treatment with autophagy inhibitor chloroquine, suggesting that autophagy inhibition participated in anti-inflammatory effect. We also found that punicalagin downregulated FoxO3a expression, resulting in autophagy inhibition. Overall these results suggested that punicalagin played an important role in the attenuation of LPS-induced inflammatory responses in RAW264.7 macrophages and that the mechanisms involved downregulation of the FoxO3a/autophagy signaling pathway.

Inhibitory Effect of Delphinidin on Oxidative Stress Induced by H₂O₂ in HepG2 Cells: Chronic liver diseases (CLDs) are correlated with oxidative stress induced by the accumulation of intracellular reactive oxygen species (ROS). In this study, we employed HepG2, a human liver carcinoma cell line containing many antioxidant enzymes, to explore the function of delphinidin against oxidative stress induced by H₂O₂ and to provide scientific data of the molecular mechanism. Cells were pretreated with different concentrations of delphinidin (10μmol/L, 20μmol/L, and 40μmol/L) for 2h before treatment with 750μM H₂O₂ for 1h. The results showed that H₂O₂ decreased the survival rate of HepG2 cells and increased the level of ROS, but delphinidin pretreatment could possess the opposite result. At the same time, the expression of Nrf2 was enhanced by the delphinidin pretreatment. This was because delphinidin promoted Nrf2 nuclear translocation and inhibited its degradation, which led to the increase expression of antioxidant protein HO-1 (Nrf2-related phase II enzyme heme oxygenase-1). Besides, we found that delphinidin could significantly alleviate the reduction of Nrf2 protein levels and the accumulation of intracellular ROS levels in Nrf2 knockdown HepG2 cells. In conclusion, our study suggested that delphinidin, as an effective antioxidant, protected HepG2 cells from oxidative stress by regulating the expression of Nrf2/HO-1.

Gouty arthritis serves as an acute reaction initiated by the deposition of monosodium urate (MSU) crystals around the joints. In this study, the anti-inflammatory effects of phytochemical β-caryophyllene on MSU crystal-induced acute gouty arthritis in vivo and in silico were explored. Through bioinformatics methods and molecular docking, it screened the specific influence pathway of β-caryophyllene on gout. Certain methods including enzyme-linked immunosorbent assay, western blotting, and immunohistochemical staining were adopted to quantify. β-caryophyllene significantly reduced inflammation and function of ankle joints in MSU Crystals-induced gouty arthritis rats, while decreasing serum cytokine levels. Furthermore, it inhibited the expressions of NLRP3, Caspase-1, ASC, TLR4, MyD88, p65, and IL-1β in the synovial tissue so as to reduce inflammation and protect ankle joints’ function. A new research approach in which β-caryophyllene treatment to acute attacks of gout is provided through the research results.

Approximately 80% of global tuberculosis (TB) cases occur in low-resource settings, with little opportunity for TB control. We hypothesized that the rapid increase in smartphone users and advances in digital technology would render bracelet-based applications possible; specifically, that bracelet- and self-directed observational therapy (BSDOT) can be used by patients with TB to ensure adherence to TB medication regimens and by basic village physicians to monitor care. This will ultimately allow TB to be controlled in low-resource environments. This study will have three phases: development of a bracelet capable of storing pills and recording adherence to medication regimens; creation of a BSDOT smartphone application capable of supporting reminders to patients and health care interactions between patients and village physicians; and performance of a cluster randomized controlled trial in Hunan Province, China. Patients in the intervention group will receive free bracelets and smartphones, and their daily medication intake will be directed by the smartphones; the control group will receive no intervention. The primary outcome will be the TB treatment result as defined by the World Health Organization (WHO) as follows: Cured, Treatment completed, Treatment failed, Died, Lost to follow-up, Not evaluated, or Treatment success. The secondary outcome will be treatment adherence, defined as the percentage of patients receiving TB treatment who missed fewer than 5% of doses. We will also assess self-reported adherence using the Morisky, Green, and Levine Adherence Scale (MGLS) and evaluate respondents’ knowledge about TB and quality of life. A regression model will be used to explore whether the interventions improve drug adherence and other outcome measures. This will be a powerful means by which to strengthen TB control and prevent TB, especially multidrug-resistant epidemics of the disease. In addition, our novel smartphone-based tool can be readily adopted for use in low-resource remote environments with limited health care facilities and few economic assets. The protocol has been approved by the Ethics Committee of Xiangya School of Public Health, Central South University (reference number: XYGW-2016-14). Chinese Clinical Trial Registry, ID: ChiCTR-IOR-16008424 . Registered on 5 June 2016.

Exposure to silica nanoparticles (SNPs) causes lung fibrosis and threatens human health. However, it is unknown if low-dose radiation (LDR) exposure exacerbates SNP-induced lung dysfunction. Thus, the aim of our study was to determine the combined effect of SNPs and LDR on lung fibrosis and elucidate the potential mechanisms involved. We used SNP-induced A549 cells and mouse models and detected gut microbiota alteration by 16S rDNA amplicon sequencing. Additionally, lung fibrosis-related parameters were also detected using hematoxylin and eosin (H&E) staining, immunofluorescence (IF) staining, qRT-PCR, and western blot analysis. The histopathological and IF staining assays illustrated that co-exposure of mice to SNPs and LDR had a significant deleterious effect on both lung function and lung fibrosis, in comparison to exposure to SNPs or LDR alone. Furthermore, the abundance of Bacteroidetes significantly increased while that of Firmicutes significantly decreased following co-exposure to SNPs and LDR. Mechanistically, the Notch cascade was activated in chronically SNP-exposed mice with lung fibrosis and A549 cells. Additionally, the Notch pathway-associated proteins showed increased expression levels in the lungs following SNP exposure both in vivo and in vitro. Notably, SNP-induced dysbiosis of the gut microbiota promoted lung epithelial damage by triggering the Notch pathway, resulting in SNP-induced lung fibrosis. However, oral administration of probiotics protected the mice from SNP-induced lung injury. Our results strongly indicate that the activation of the gut microbiota-dependent Notch pathway in response to co-exposure to SNPs and LDR results in lung epithelial injury in vivo. Probiotics supplementation is a potential way to protect against SNP- and LDR-induced lung fibrosis.