The Extracts Derived from Artemisia japonica Thunb. Leaves Mitigate Oxidative Stress and Inflammatory Response Induced by LPS in RAW264.7 Cells through Modulation of the Nrf2/HO-1 Signaling Pathway.

Artemisia japonica Thunb. has been used as a traditional Chinese medicine and a vegetable for thousands of years in China. However, there are few reports on the chemical composition and biological activity of its leaves. Thus, this study aimed to evaluate the chemical composition, antioxidant and anti-inflammatory effects of water extracts of A. japonica leaves and their underlying mechanisms. A total of 48 compounds were identified in the water extract using UPLC-QTOF-MS2 analysis, with phenolic acids, particularly chlorogenic acid compounds, being the predominant components. The ethyl acetate fraction (EAF) contained most of the total phenolic content (385.4217 mg GAE/g) and displayed superior antioxidant capacity with the IC50DPPH•, IC50ABTS•+, and OD0.5reducing power at 10.987 µg/mL, 43.630 µg/mL and 26.883 µg/mL, respectively. Furthermore, EAF demonstrated potent antioxidant and anti-inflammatory effects in LPS-induced RAW264.7 cells by upregulating the Nrf2/HO-1 signal pathway. These findings highlight that A. japonica leaves possess remarkable abilities to mitigate inflammation and oxidative stress, suggesting their potential utilization as medicinal agents and food additives for promoting human health.

Effect fraction of Bletilla striata (Thunb.) Reichb.f. alleviates LPS-induced acute lung injury by inhibiting p47phox/NOX2 and promoting the Nrf2/HO-1 signaling pathway.

BACKGROUND & AIMS: The effect fraction of Bletilla striata (Thunb.) Reichb.f. (EFBS), a phenolic-rich extract, has significant protective effects on lipopolysaccharide (LPS)-induced acute lung injury (ALI), but its composition and molecular mechanisms are unclear. This study elucidated its chemical composition and possible protective mechanisms against LPS-induced ALI from an antioxidant perspective. METHODS: EFBS was prepared by ethanol extraction, enriched by polyamide column chromatography, and characterized using ultra-performance liquid chromatography/time-of-flight mass spectrometry. The LPS-induced ALI model and the RAW264.7 model were used to evaluate the regulatory effects of EFBS on oxidative stress, and transcriptome analysis was performed to explore its possible molecular mechanism. Then, the pathway by which EFBS regulates oxidative stress was validated through inhibitor intervention, flow cytometry, quantitative PCR, western blotting, and immunofluorescence techniques. RESULTS: A total of 22 compounds in EFBS were identified. The transcriptome analyses of RAW264.7 cells indicated that EFBS might reduce reactive oxygen species (ROS) production by inhibiting the p47phox/NADPH oxidase 2 (NOX2) pathway and upregulating the nuclear factor erythroid 2-related factor 2 (Nrf2)/heme oxygenase-1 (HO-1) pathway. Both in vitro and in vivo data confirmed that EFBS significantly inhibited the expression and phosphorylation of p47phox protein, thereby weakening the p47phox/NOX2 pathway and reducing ROS production. EFBS significantly increased the expression of Nrf2 in primary peritoneal macrophages and lung tissue and promoted its nuclear translocation, dose-dependent increase in HO-1 levels, and enhancement of antioxidant activity. In vitro, both Nrf2 and HO-1 inhibitors significantly reduced the scavenging effects of EFBS on ROS, further confirming that EFBS exerts antioxidant effects at least partially by upregulating the Nrf2/HO-1 pathway. CONCLUSIONS: EFBS contains abundant phenanthrenes and dibenzyl polyphenols, which can reduce ROS production by inhibiting the p47phox/NOX2 pathway and enhance ROS clearance activity by upregulating the Nrf2/HO-1 pathway, thereby exerting regulatory effects on oxidative stress and improving LPS-induced ALI.

Global proteome and lysine succinylation analyses provide insights into the secondary metabolism in Salvia miltiorrhiza.

Danshen, belongs to the Lamiaceae family, and its scientific name is Salvia miltiorrhiza Bunge. It is a valuable medicinal plant to prevent and treat cardiovascular and cerebrovascular diseases. Lysine succinylation, a widespread modification found in various organisms, plays a critical role in regulating secondary metabolism in plants. The hairy roots of Salvia miltiorrhiza were subject to proteomic analysis to identify lysine succinylation sites using affinity purification and HPLC-MS/MS in this investigation. Our findings reveal 566 lysine succinylation sites in 348 protein sequences. We observed 110 succinylated proteins related to secondary metabolism, totaling 210 modification sites. Our analysis identified 53 types of enzymes among the succinylated proteins, including phenylalanine ammonia-lyase (PAL) and aldehyde dehydrogenase (ALDH). PAL, a crucial enzyme involved in the biosynthesis of rosmarinic acid and flavonoids, displayed succinylation at two sites. ALDH, which participates in the phenylpropane metabolic pathway, was succinylated at 8 eight sites. These observations suggest that lysine succinylation may play a vital role in regulating the production of secondary metabolites in Salvia miltiorrhiza. Our study may provide valuable insights for further investigation on plant succinylation, specifically as a reference point. SIGNIFICANCE: Salvia miltiorrhiza Bunge is a valuable medicinal plant that prevents and treats cardiovascular and cerebrovascular diseases. Lysine succinylation plays a critical role in regulating secondary metabolism in plants. The hairy roots of Salvia miltiorrhiza were subject to proteomic analysis to identify lysine succinylation sites using affinity purification and HPLC-MS/MS in this investigation. These observations suggest that lysine succinylation may act as a vital role in regulating the production of secondary metabolites in Salvia miltiorrhiza. Our study may provide valuable insights for further investigation on succinylation in plants, specifically as a reference point.

Chemical Composition, Antitumor Properties, and Mechanism of the Essential Oil from Plagiomnium acutum T. Kop.

Plagiomnium acutum T. Kop. (P. acutum) has been used as a traditional Chinese medicine for thousands of years to treat cancer but lacks evidence. The objective of this work was to reveal the chemical composition of P. acutum essential oil (PEO) and explore its potential antitumor activity and molecular mechanism. PEO was prepared by the simultaneous distillation-extraction method and characterized by gas chromatography/mass spectroscopy. CCK8 assay, flow cytometry, western blot, and immunofluorescence techniques were used to analyze the effects and mechanism of PEO against cancer cells. A total of 74 constituents of PEO were identified, with diterpenes (26.5%), sesquiterpenes (23.89%), and alcohols (21.81%) being the major constituents. Two terpenoids, selina-6-en-4-ol and dolabella-3,7-dien-18-ol, were detected in PEO for the first time. PEO showed significant cell growth inhibitory activity on HepG2 and A549 cells by blocking the G1 phase and inducing apoptosis, which may be attributed to its upregulation of p21Cip1 and p27Kip1 proteins and interference with mitochondrial membrane potential effect. Dolabella-3,7-dien-18-ol accounts for 25.5% of PEO and is one of the main active components of PEO, with IC50 values in HepG2 and A549 cells of (25.820 ± 0.216) µg/mL and (23.597 ± 1.207) µg/mL, respectively. These results confirmed the antitumor medicinal value of P. acutum and showed great application potential in the pharmaceutical industry.

Integrated microbiology and metabolomics analysis reveal responses of soil microorganisms and metabolic functions to phosphorus fertilizer on semiarid farm.

Localized fertilization of phosphorus has potential benefits in achieving higher crop productivity and nutrient use efficiency, but the underlying biological mechanisms of interactions between soil microorganisms and related metabolic cycle remain largely to be recognized. Here, we combined microbiology with non-target metabolomics to explore how P fertilizer levels and fertilization patterns affect wheat soil microbial communities and metabolic functions based on high-throughput sequencing and UPLC-MS/MS platforms. The results showed P fertilizer decreased the diversity of bacterial 16S rRNA genes and fungal ITS genes, and it did significantly change both soil bacterial and fungal overall community structures and compositions. The P levels and patterns also interfered with complexity of soil bacterial and fungal symbiosis networks. Moreover, metabolomics analysis showed that P fertilizer significantly changed soil metabolite spectrum, and the differential metabolites were significantly enriched to 7 main metabolic pathways, such as arginine and proline metabolism, biosynthesis of plant hormones, amino acids, plant secondary metabolites, and alkaloids derived from ornithine. Additionally, microbes also were closely related to the accumulation of metabolites through correlation analysis. Our results indicated that localized appropriate phosphorus fertilizer plays an important role in regulating soil microbial metabolism, and their interactions in soil providing valuable information for understanding how the changed phosphorus management practices affect the complex biological processes and the adaption capacity of plants to environments.

Effects of different mulching and fertilization on phosphorus transformation in upland farmland.

In the present study, the impact of different soil surface mulching, fertilization on phosphorus mineralization and bio-availability of spring maize at various growth stages and soil layers (0-20 and 20-40 cm soil layer) were evaluated. The results indicated that the contents of total P and Olsen-Phosphorus (Olsen-P) in the soils of 0-20 cm soil layer were significantly higher than those in the 20-40 cm soil layer at different stages. The addition of organic fertilizer significantly increased the soil total P and Olsen-P content in the 0-20 cm soil layer. The different surface mulching, no mulching (NM), gravel mulching (GM) and film mulching (FM) were significantly affected by the content of Olsen-P in both soil layers during the critical growth period of spring maize. The Ca10-P contents in both soil layers were the maximum in terms of the inorganic phosphorus content in soils with different surface mulching and different fertilization. Surface mulching significantly affected the transformation of inorganic phosphorus in different soil layers of dry-land farmland, and accelerated the increase of Ca2-P content (first phosphorus source) in 0-20 cm soil layer by GM and FM. In addition, phosphorus combined with inorganic nitrogen fertilizer increased Ca8-P (second Olsen-P source) to a certain extent, and reduced the relative content of Ca2-P (first phosphorus source). Compared with phosphate (P), nitrogen and phosphorus (NP) treatments, manure and nitrogen and phosphorus (MNP) treatments increased the contents of Ca2-P (first phosphorus source) and Ca8-P (second effective phosphorus source), while it reduced the insoluble phosphorus source (O-P) content.

TMP prevents retinal neovascularization and imparts neuroprotection in an oxygen-induced retinopathy model.

PURPOSE: To evaluate the effects of tetramethylpyrazine (TMP) on retinal neovascularization (NV) and neuroprotection in an oxygen-induced retinopathy (OIR) model. METHODS: Neonatal C57BL/6J mice were subjected to 75% oxygen from postnatal day 7 (P7) to P12 and then returned to room air. TMP (200 mg/kg) or normal saline was given daily from P12 to P17. Immunostaining, HE staining, TUNEL assay, and RT-PCR were used to assess the effects of TMP on retinal neurovascular repair. RESULTS: TMP effectively prevented pathologic NV and accelerated physiologic revascularization by enhancing the formation of endothelial tip cells at the edges of the repairing capillary networks and preserving the astrocytic template in the avascular retina. TMP also prevented morphologic changes and significantly decreased TUNEL-positive cells in the avascular retina by rescuing neurons such as amacrine, rod bipolar, horizontal, and Müller cells. In TMP-treated mice retinas, there was a less obvious loss of amacrine cell bodies and their distinct bands; the number of both rod bipolar and horizontal cell bodies, as well as the density of their dendrites in the outer plexiform layer, was greater than that in OIR control mice. TMP not only decreased the loss of alignment of Müller cell bodies and distortion of processes but reduced the reactive expression of GFAP in Müller cells. Furthermore, HIF-1α and VEGF mRNA expression were downregulated in TMP-treated mice retinas. CONCLUSIONS: TMP improved neurovascular recovery by preventing NV and protecting retinal astroglia cells and neurons from ischemia-induced cell death partially due to its downregulation of HIF-1α and VEGF mRNA expression.

Protective effect of a polyphenolic rich extract from Magnolia officinalis bark on influenza virus-induced pneumonia in mice.

ETHNOPHARMACOLOGICAL RELEVANCE: Magnolia officinalis bark is used in traditional Chinese medicine for the treatment of cough, colds, fever, chronic bronchitis and stomach ailments. AIM OF THE STUDY: To investigate therapeutic effects of polyphenol rich extract from M. officinalis bark (MPE) in influenza virus A-infected mice, and to provide evidence for the inflammation response and immunomodulatory potential during infection. MATERIALS AND METHODS: Mice were infected with influenza virus A (IVA) and MPE at doses of 10 and 20mg/kg were orally administrated daily for 5 days after challenge. The levels of serum L-6 and TNF-α were determined by ELISA while protein expressions of NF-κB and TLR3 were detected by western blotting analysis. RESULTS: MPE exhibited significant therapeutical effects on reducing levels of serum NO, IL-6 and TNF-α, inhibiting pneumonia, decreasing lung viral titers and sensitizing IVA-induced apoptosis through down-regulation of NF-κB and TLR3 protein expression in the lung tissue of IVA-infected mice. CONCLUSIONS: MPE could exhibit preventive and therapeutical effects on IVA-infected mice as a suppressor of the production of inflammatory mediators, NO and pro-inflammatory cytokines, TNF-α and IL-6. These effects appeared to be mediated, at least in part, by an inhibition of TLR3 and NF-κB activation. Therefore, MPE could provide a safe and effective therapeutic approach for influenza and its subsequent viral pneumonia.

[Effects of spatial coupling of watering and fertilization on winter wheat photosynthetic characteristics and grain yield].

A soil column experiment was conducted with a fertile Eum-Orthic Anthrosols to study the effects of spatial coupling of watering and nitrogen (N)- and phosphorus (P) fertilization on the photosynthetic characteristics and grain yield of winter wheat. The column consisted of three layers, each layer being 30 cm in thickness and with an underlying 2 cm layer of coarse sand to obstruct water- and nutrient exchange. The results showed that relative content of chlorophyll (SPAD), net photosynthetic rate (P(n)), and grain yield were significantly lower in the treatment of 0-30 cm soil drought and 30-90 cm soil wetness (D) than in that of 0-90 cm soil wetness (W). In these two watering treatments, a combined application of N and P got the highest SPAD, P(n) and grain yield, followed by applying P, and N. The SPAD, P(n) and grain yield were significantly higher when the fertilizer N was applied in 0-90 cm layer than in 0-30, 30-60 and 60-90 cm layers, respectively. No significant differences in SPAD, P(n) and grain yield were observed when fertilizer P was applied in 0-90 cm layer and in 0-30 cm layer. In treatment W, the SPAD, P(n) and grain yield were the highest when N and P were applied in 0-90 cm layer, had no significant differences with those when the N and P were applied in 0-30 cm layer, but were significantly higher than those when the fertilization was carried out in 30-60 and 60-90 cm layers. In treatment D, no significant difference in SPAD was observed when the N and P were applied in different layers, but P(n) and grain yield were significant higher when the N and P were applied in 0-90 cm than in 30-60 cm layer, and in 30-60 cm than in 60-90 cm layer. It was concluded that whether in treatment W or in treatment D, a combined application of N and P should be carried out in 0-30 cm soil layer.

[Effect of plastic film mulching on soil microbial biomass in spring wheat field in semi-arid loess area].

This paper studied the effect of different periods of plastic film mulching (M0-no mulching, M30-mulching for 30 days, M60-mulching for 60 days, and M-mulching for whole growth period) on soil microbial biomass carbon (SMBC) of spring wheat field in semi-arid loess plateau. The mean SMBC in 1999 and 2000 was 335.3 and 259.3 mg.kg-1 dry soil, respectively. It was 29.3% higher in 1999 than in 2000. The highest SMBC was recorded at the harvest stage in M treatment for the two years. In 1999, a wet year with more rainfall, the SMBC of M60 and M treatments was significantly higher than those of M0 and M30 in the mid-period of growth, reached its top at the end of the growth period. The highest grain yield was also achieved in M60. It was a dry year in 2000, but rainfall was rich in the latter growth period of spring wheat. SMBC increased at the beginning period of growth, and did not increase during the mid-growth period. It increased again during the latter period of growth, and showed a significant difference among the treatments. At the harvest stage of 2000, SMBC in M0 was the highest among all the treatments. It was similar between M and M60, and lower than that of M30. In the two years, the ratio of C/N ranged between 7.732-9.042, being lower than the threshold of 11.3, and the ratio of C/P was 300.8-719.6, being higher than the threshold of 300. The two parameters showed that the increase of SMBC was inhibited because of the competition of soil available nutrients between soil microbes and crops. These indicated that soil organic matter content was so shortage that it became the key factor to restrict crop productivity. Under this condition, increasing crop productivity through the input of chemical fertilizers would conceal the problem of soil degradation, and result in a further decrease of soil quality. A long term plastic film mulching would make the problem more serious.