ABSTRACT
Aim To explore the effect of berberine (B E) on RSV infected HEp-2 cells and the related mechanism. Methods HEp-2 cells were infected with RSV and treated with BE. Cell viability was assessed using the CCK-8 assay. Protein expression levels of NLRP3, ASC, caspase-1, PINK1, Parkin, Beclinl, p62, LC3 I,LC3 II,and BNIP3 in HEp-2 cells were detected by Western blot. The secretion level of IL-1 p in HEp-2 cells was measured using ELISA. Apoptosis rate and mitochondrial membrane potential of HEp-2 cells were examined by flow cytometry. Mitochondrial ROS (mtROS) in HEp-2 cells was detected through MitoSOX staining. Colocalization of mitochondria and autophagosomes in HEp-2 cells was investigated using immunofluorescence staining. Cyclosporin A was used for validation experiments. Results BE could significantly improve the activity of RSV-infected HEp-2 cells,reduce the apoptosis rate (P < 0. 05), and decrease the activation level of NLRP3 inflammasomes and IL-lp level (P <0. 05); BE improved mitochondrial function by increasing mitochondrial membrane potential and ATP levels,and reduced mtROS. BE significantly promoted the colocalization of mitochondria-autophagosome in RSV infected cells, inducing PINK1/ Parkin and BNIP3 to mediate mitochondrial autophagy; cyclosporine A aggravated RSV infection. Conclusions BE has protective effects on HEp-2 cells infected by RSV. The mechanism may be related to the inhibitory effect of BE on the production of mtROS and the activation of NLRP3 inflammasomes by inducing PINK1/ Parkin and BNIP3-mediated mitochondrial autophagy.
ABSTRACT
A anaerobic hydrogen-producing strain HR-1 was isolated from compost. Phylogenetic analysis based on 16S rRNA sequence similarity indicates that strain HR-1 is the closest relative to Clostridium ace- tobutylicum ATCC 824, with the similarity of 96%. Biological characteristics and phylogenetic analysis of 16S rRNA gene indicate that HR-1 is a new species named Clostridium sp. HR-1. Cells are Gram-positive, mobile rod-shaped. Spores and flagellums were no observed. Temperature range for growth is 10?C to 45?C (optimum temperature 37?C~39?C), and range pH for growth is 4.0 to 10.0 (optimum pH 7.5~8.0). H2, CO2, acetate, butyrate and a little ethanol are the end products of PYG fermentation. Strain HR-1 has the ability to use organic nitrogen and inorganic nitrogen sources for growth and hydrogen production, and yeast extract is the optimum nitrogen source for hydrogen production. Strain HR-1 produces hydrogen from xylose (3 g/L) at 37?C and initial pH 6.5, the hydrogen yields and maximal hydrogen production rate are 1.84 mol-H2/mol-xylose and 10.52 mmol-H2/h?g-cdw, respectively. Strain HR-1 is able to utilize glucose, galactose, fructose, mannose and cellobiose for hydrogen production and the hydrogen yields from glucose is 2.36 mol-H2/mol-glucose.