Lignin from different biomasses possess biological antioxidation and antimicrobial activities, which depend on the number of functional groups and the molecular weight of lignin. In this work, organosolv fractionation was carried out to prepare the lignin fraction with a suitable structure to tailor excellent biological activities. Gel permeation chromatography (GPC) analysis showed that decreased molecular weight lignin fractions were obtained by sequentially organosolv fractionation with anhydrous acetone, 50% acetone and 37.5% hexanes. Nuclear magnetic resonance (NMR) results indicated that the lignin fractions with lower molecular weight had fewer substructures and a higher phenolic hydroxyl content, which was positively correlated with their antioxidation ability. Both of the original lignin and fractionated lignins possessed the ability to inhibit the growth of Gram-negative bacteria (Escherichia coli and Salmonella) and Gram-positive bacteria (Streptococcus and Staphylococcus aureus) by destroying the cell wall of bacteria in vitro, in which the lignin fraction with the lowest molecular weight and highest phenolic hydroxyl content (L3) showed the best performance. Besides, the L3 lignin showed the ability to ameliorate Escherichia coli-induced diarrhea damages of mice to improve the formation of intestinal contents in vivo. These results imply that a lignin fraction with a tailored structure from bamboo lignin can be used as a novel antimicrobial agent in the biomedical field.
Microplastics (MPs) generally refer to the plastic fragments or particles smaller than 5 mm in diameter, which are closely concerned due to their widespread presence in the environment. Recent studies have shown that MPs have a serious threat on the reproductive health of organisms. Pigs are often selected as the model animals because of their high similarity to human tissues and organs. However, there are no reports on the effects and mechanisms of MPs exposure on swine germ cells. In the present study, we established swine testis (ST) cell models exposed to 250, 500, and 1000 μg/ml polystyrene microplastics (PS‐MPs, 1–10 μm), respectively. The findings revealed that PS‐MPs reduced cell viability dose‐dependently. Acridine orange/ethidium bromide staining and flow cytometry results indicated the occurrence of apoptosis and necrosis in ST cells under PS‐MPs exposure, and the expression changes of relevant marker genes (B‐cell lymphoma‐2, Bcl‐2 Associated X, Caspase‐3, Caspase‐9, Receptor‐interacting protein kinase 1, Receptor‐interacting protein kinase 3, Mixed lineage kinase domain‐like, and Caspase‐8) were clarified via quantitative real‐time PCR and western blot. Further mechanistic studies found that PS‐MPs treatment induced excessive intracellular reactive oxygen species (ROS) production, which promoted the phosphorylation of mitogen‐activated protein kinase (MAPK) pathway‐related genes (P38, c‐Jun N‐terminal kinase, extracellular regulated protein kinases) and activated the downstream gene hypoxia‐inducible factor (HIF1α). In conclusion, our study suggests that PS‐MPs treatment causes apoptosis and necroptosis in ST cells via ROS/MAPK/HIF1α signaling pathway.
Tetrabromobisphenol A (TBBPA) is a widely used industrial brominated flame retardant, which can endanger animal and human health, including cytotoxicity, endocrine disruption, reproductive toxicity and so on. Melatonin (MT) is a noteworthy free radical scavenger and an antioxidant to alleviate oxidative stress. To investigate the cytotoxic of TBBPA on swine testis cells (ST cells), as well as the antagonistic effect of MT, we established TBBPA exposure and MT antagonistic models, used flow cytometry and AO/EB staining methods to detect apoptosis and necroptosis, used DCFH-DA method to examine the content of reactive oxygen species (ROS) and investigated the expression of associated genes using RT-PCR and Western blot. According to our findings, TBBPA exposure induced cell death in ST cells. TBBPA increased ROS levels, thus increasing PTEN expression and decreasing PI3K and AKT expression. Apoptosis-related factors (Caspase-3, Bax, Cyt-c, and Caspase-9) and necroptosis-related factors (RIPK1, RIPK3, and MLKL) were considerably elevated, in addition to the reduced expression of BCL-2 and Caspase-8. We also found that MT inhibited apoptosis and necroptosis in TBBPA-induced ST cells and effectively resolved the abnormal expression of related signaling pathways. In summary, the above results indicate that MT alleviates the disorder of PTEN/PI3K/AKT signaling pathway via inhibiting ROS overproduction, thereby mitigating apoptosis and necroptosis caused by TBBPA. This research provides a theoretical basis for further understanding of the toxicity of TBBPA and the detoxification of MT against environmental toxics.
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