Salicylic acid (SA) is an elicitor to induce the biosynthesis of secondary metabolites in plant cells. Hydrogen peroxide (H2O2) plays an important role as a key signaling molecule in response to various stimuli and is involved in the accumulation of secondary metabolites. However, the relationship between them is unclear and their synergetic functions on accumulation of secondary metabolites are unknown. In this paper, the roles of SA and H2O2 in rosmarinic acid (RA) production in Salvia miltiorrhiza cell cultures were investigated. The results showed that SA significantly enhanced H2O2 production, phenylalanine ammonia-lyase (PAL) activity, and RA accumulation. Exogenous H2O2 could also promote PAL activity and enhance RA production. If H2O2 production was inhibited by NADPH oxidase inhibitor (IMD) or scavenged by quencher (DMTU), RA accumulation would be blocked. These results indicated that H2O2 is secondary messenger for signal transduction, which can be induced by SA, significantly and promotes RA accumulation.

Expansins, cell-wall loosening proteins, play an important role in plant growth and development and abiotic stress tolerance. Ammopiptanthus nanus (A. nanus) is an important plant to study to understand stress resistance in forestry. In our previous study, two α-type expansins from A. nanus were cloned and named AnEXPA1 and AnEXPA2. In this study, we found that they responded to different abiotic stress and hormone signals. It suggests that they may play different roles in response to abiotic stress. Their promoters show some of the same element responses to abiotic stress and hormones, but some special elements were identified between the expansins that could be essential for their expression. In order to further testify the reliability of the above results, we conducted an analysis of β-glucuronidase (GUS) dyeing. The analysis showed that AnEXPA1 was only induced by cold stress, whereas AnEXPA2 responded to hormone induction. AnEXPA1 and AnEXPA2 transgenic Arabidopsis plants showed better tolerance to cold and drought stresses. Moreover, the ability to scavenge reactive oxygen species (ROS) was significantly improved in the transgenic plants, and expansin activity was enhanced. These results suggested that AnEXPA1 and AnEXPA2 play an important role in the response to abiotic stress. Our research contributes to a better understanding of the regulatory network of expansins and may benefit agricultural production.

Chlorogenic acid (CGA), a phenylpropanoid derived from Eucommia ulmoides Oliver, has been shown to exhibit potent cytotoxic and anti-proliferative activities against several human cancers. However, the effects of CGA on hepatocellular carcinoma (HCC) and the underlying mechanisms have not been intensively studied. In this study, the CGA treatment effects on the viability of human hepatoma cells were investigated by MTT assay. Our data showed that CGA could dose-dependently inhibit the activity of human hepatoma cells Hep-G2 and Huh-7, but did not affect the activity and growth of normal human hepatocyte QSG-7701. The genes and pathways influenced by CGA treatment were explored by RNA sequencing and bioinformatics analysis, which identified 323 differentially expressed genes (DEGs) involved in multiple pharmacological signaling pathways such as MAPK, NF-κB, apoptosis and TGF-β signaling pathways. Further analyses by real-time quantitative PCR, Western blot and flow cytometry revealed that CGA effectually suppressed the noncanonical NF-κB signaling pathway, meanwhile it activated the mitochondrial apoptosis of HCC by upregulation of the BH3-only protein Bcl-2 binding component 3 (BBC3). Our findings demonstrated the potential of CGA in suppressing human hepatoma cells and provided a new insight into the anti-cancer mechanism of CGA.

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