Towards Better Sinomenine-Type Drugs to Treat Rheumatoid Arthritis: Molecular Mechanisms and Structural Modification.

Sinomenine is the main component of the vine Sinomenium acutum. It was first isolated in the early 1920s and has since attracted special interest as a potential anti-rheumatoid arthritis (RA) agent, owing to its successful application in traditional Chinese medicine for the treatment of neuralgia and rheumatoid diseases. In the past few decades, significant advances have broadened our understanding of the molecular mechanisms through which sinomenine treats RA, as well as the structural modifications necessary for improved pharmacological activity. In this review, we summarize up-to-date reports on the pharmacological properties of sinomenine in RA treatment, document their underlying mechanisms, and provide an overview of promising sinomenine derivatives as potential RA drug therapies.

Heterologous Biosynthesis of Health-Promoting Baicalein in Lycopersicon esculentum.

Baicalein is a valuable flavonoid isolated from the medicinal plant Scutellaria baicalensis Georgi, which exhibits intensive biological activities, such as anticancer and antiviral activities. However, its production is limited in the root with low yield. In this study, In-Fusion and 2A peptide linker were developed to assemble SbCLL-7, SbCHI, SbCHS-2, SbFNSII-2 and SbCYP82D1.1 genes driven by the AtPD7, CaMV 35S and AtUBQ10 promoters with HSP, E9 and NOS terminators, and were used to engineer baicalein biosynthesis in transgenic tomato plants. The genetically modified tomato plants with this construct synthesized baicalein, ranging from 150 ng/g to 558 ng/g FW (fresh weight). Baicalein-fortified tomatoes have the potential to be health-promoting fresh vegetables and provide an alternative source of baicalein production, with great prospects for market application.

[Pharmacodynamics and intestinal flora research on different processed products of Puerariae Lobatae Radix and Puerariae Thomsonii Radix].

This study aims to explore the pharmacodynamic differences of Puerariae Lobatae Radix(PLR), Puerariae Thomsonii Radix(PTR) and their different processed products and the influences of these medical materials on the diversity of intestinal flora. The Sennae Folium-induced diarrhea model, streptozotocin(STZ)-induced diabetes model and L-nitro-arginine methyl ester(L-NAME)-induced hypertension model were used to compare the pharmacodynamic differences in anti-diarrhea, blood glucose reduction and blood pressure lowering among raw, roasted and vinegar-processed PLR and PTR. The effects of raw and processed PLR and PTR on intestinal flora diversity of rats were evaluated by 16 S rDNA high-throughput sequencing. The roasted PLR and PTR performed better in anti-diarrhea, especially the former. PLR and its processed products all presented the efficacy of reducing blood glucose, and the vinegar-processed PLR was the most outstanding. The raw PTR was not that effective in reducing blood glucose, whereas its efficacy was improved after roasting and vinegar processing. Both PLR and PTR were capable of lowering blood pressure to a certain extent, and PLR is superior to PTR in this aspect. Further, the vinegar-processed PLR showed the best effect. The diversity of intestinal flora was different among rats to which different products of PLR and PTR were administered. The roasted PLR led to the highest abundance of Lactobacillus, which was closely related to its best antidiarrheal effect. The highest abilities of vinegar-processed PLR to lower blood glucose and blood pressure were associated with the high abundance of Blautia and Prevotella_9. This study lays a foundation for elucidating the processing mechanisms of PLR and PTR and provides a basis for their further development and application.

Development of an efficient transient expression system for Siraitia grosvenorii fruit and functional characterization of two NADPH-cytochrome P450 reductases.

Siraitia grosvenorii (Luo hanguo or monk fruit) is a valuable medicinal herb for which the market demand has increased dramatically worldwide. As promising natural sweeteners, mogrosides have received much attention from researchers because of their extremely high sweetness and lack of calories. Nevertheless, owing to the absence of genetic transformation methods, the molecular mechanisms underlying the regulation of mogroside biosynthesis have not yet been fully elucidated. Therefore, an effective method for gene function analysis needs to be developed for S. grosvenorii fruit. As a powerful approach, transient expression has become a versatile method to elucidate the biological functions of genes and proteins in various plant species. In this study, PBI121 with a ß-glucuronidase (GUS) marker and tobacco rattle virus (TRV) were used as vectors for overexpression and silencing, respectively, of the SgCPR1 and SgCPR2 genes in S. grosvenorii fruit. The effectiveness of transient expression was validated by GUS staining in S. grosvenorii fruit, and the expression levels of SgCPR1 and SgCPR2 increased significantly after infiltration for 36 h. In addition, TRV-induced gene silencing suppressed the expression of SgCPR1 and SgCPR2 in S. grosvenorii fruit. More importantly, the production of the major secondary metabolites mogrol, mogroside IIE (MIIE) and mogroside III (MIII) was activated by the overexpression of SgCPR1 and SgCPR2 in S. grosvenorii fruit, with levels 1-2 times those in the control group. Moreover, the accumulation of mogrol, MIIE and MIII was decreased in the SgCPR1 and SgCPR2 gene silencing assays. Therefore, this transient expression approach was available for S. grosvenorii fruit, providing insight into the expression of the SgCPR1 and SgCPR2 genes involved in the mogroside biosynthesis pathway. Our study also suggests that this method has potential applications in the exploration of the molecular mechanisms, biochemical hypotheses and functional characteristics of S. grosvenorii genes.

Effects of Forchlorfenuron on the Morphology, Metabolite Accumulation, and Transcriptional Responses of Siraitia grosvenorii Fruit.

Siraitia grosvenorii fruit, called luo-han-guo (LHG), have been used as a traditional Chinese medicine (TCM) and dietary supplements for many years. Mogrosides, the main bioactive ingredients in LHG, are commercially available worldwide as a non-sugar-based and noncaloric sweetener. However, the production cannot meet the increasing market demand because of the low content of mogrosides and the small size of LHG. Therefore, some advanced technologies have been applied for improving the quality of LHG. Forchlorfenuron (CPPU), a plant growth regulator, is widely applied to promote plant yield and the secondary metabolite synthesis. Here, the content of nine mogrosides and three intermediates in LHG that were treated with three different concentrations of CPPU were determined by LC-MS/MS and GC-MS, respectively. The total content of mogrosides in LHG treated with CPPU was not enhanced, and the proportion of some main bioactive ingredients, including mogroside V (MV), were decreased relative to that of the control treatment. Morphological and cytological observations showed CPPU could make an early lignification in fruit epidermal cells, and 5 or 25 mg L-1 CPPU could inhibit LHG growth. The expression levels of 24 key genes in the mogroside biosynthesis pathway were measured and revealed that genes downregulated in upstream, and different expressions of SgUGTs would affect the accumulations and proportions of mogrosides in LHG induced by CPPU. This was the first study that applied CPPU individually on LHG, and assessed effects of CPPU on the morphology, the accumulation of metabolites, and expression profiles of 24 structural genes. The CPPU effects on LHG were undesirable, including development inhibition and the decrease of main mogroside content. These will provide guidance for the rational application of CPPU.

Panax notoginseng Root Cell Death Caused by the Autotoxic Ginsenoside Rg1 Is Due to Over-Accumulation of ROS, as Revealed by Transcriptomic and Cellular Approaches.

Panax notoginseng is a highly valuable medicinal herb, but its culture is strongly hindered by replant failure, mainly due to autotoxicity. Deciphering the response mechanisms of plants to autotoxins is critical for overcoming the observed autotoxicity. Here, we elucidated the response of P. notoginseng to the autotoxic ginsenoside Rg1 via transcriptomic and cellular approaches. Cellular analyses demonstrated that Rg1 inhibited root growth by disrupting the cell membrane and wall. Transcriptomic analyses confirmed that genes related to the cell membrane, cell wall decomposition and reactive oxygen species (ROS) metabolism were up-regulated by Rg1 stress. Further cellular analyses revealed that Rg1 induced ROS ([Formula: see text] and H2O2) accumulation in root cells by suppressing ascorbate peroxidase (APX) and the activities of enzymes involved in the ascorbate-glutathione (ASC-GSH) cycle. Exogenous antioxidants (ASC and gentiobiose) helped cells scavenge over-accumulated ROS by promoting superoxide dismutase (SOD) activity and the ASC-GSH cycle. Collectively, the autotoxin Rg1 caused root cell death by inducing the over-accumulation of ROS, and the use of exogenous antioxidants could represent a strategy for overcoming autotoxicity.

Identification and analysis of anthocyanin components in fruit color variation in Schisandra chinensis.

BACKGROUND: Fruit color is an important index and parameter for measuring fruit quality. As an important pigment, anthocyanin is a determinant which appears in all sorts of colors of fruits in nature. RESULTS: Color parameters were measured using a spectrometer and used as a basis to divide the materials into three groups: reddish-orange, orange and yellow. A validated high-performance liquid chromatographic-electrospray ionization-mass spectrometric method was used for the analysis of anthocyanin in Schisandra chinensis and for determining major anthocyanin components in S. chinensis fruits, i.e. cyanidin xylosyl-glucoside (CyXylGlu), cyanidin glucosyl-rutinoside (CyGluRutin), cyanidin rutinoside (CyRutin) and cyanidin xylosyl-rutinoside (CyXylRutin). The anthocyanin contents vary obviously in different colored fruits in S. chinensis. The impact of anthocyanin on coloration of fruits was investigated by multiple regression analysis between color parameters and anthocyanin components, which indicated that CyRutin is the primary cause of fruit color variation in S. chinensis. CONCLUSION: The content and type of anthocyanin determine fruit coloration in S. chinensis, laying the early foundations for systematically interpreting the mechanism of fruit coloration in S. chinensis. © 2015 Society of Chemical Industry.

Plant-plant-microbe mechanisms involved in soil-borne disease suppression on a maize and pepper intercropping system.

BACKGROUND: Intercropping systems could increase crop diversity and avoid vulnerability to biotic stresses. Most studies have shown that intercropping can provide relief to crops against wind-dispersed pathogens. However, there was limited data on how the practice of intercropping help crops against soil-borne Phytophthora disease. PRINCIPAL FINDINGS: Compared to pepper monoculture, a large scale intercropping study of maize grown between pepper rows reduced disease levels of the soil-borne pepper Phytophthora blight. These reduced disease levels of Phytophthora in the intercropping system were correlated with the ability of maize plants to form a "root wall" that restricted the movement of Phytophthora capsici across rows. Experimentally, it was found that maize roots attracted the zoospores of P. capsici and then inhibited their growth. When maize plants were grown in close proximity to each other, the roots produced and secreted larger quantities of 2,4-dihydroxy-7-methoxy-2H-1,4-benzoxazin-3(4H)-one (DIMBOA) and 6-methoxy-2-benzoxazolinone (MBOA). Furthermore, MBOA, benzothiazole (BZO), and 2-(methylthio)-benzothiazole (MBZO) were identified in root exudates of maize and showed antimicrobial activity against P. capsici. CONCLUSIONS: Maize could form a "root wall" to restrict the spread of P. capsici across rows in maize and pepper intercropping systems. Antimicrobe compounds secreted by maize root were one of the factors that resulted in the inhibition of P. capsici. These results provide new insights into plant-plant-microbe mechanisms involved in intercropping systems.