Changes in secondary metabolites contents and stress responses in Salvia miltiorrhiza via ScWRKY35 overexpression: Insights from a wild relative Salvia castanea.

Salvia castanea Diels, a close wild relative to the medicinal plant, Salvia miltiorrhiza Bunge, primarily grows in high-altitude regions. While the two species share similar active compounds, their content varies significantly. WRKY transcription factors are key proteins, which regulate plant growth, stress response, and secondary metabolism. We identified 46 ScWRKY genes in S. castanea and found that ScWRKY35 was a highly expressed gene associated with secondary metabolites accumulation. This study aimed to explore the role of ScWRKY35 gene in regulating the accumulation of secondary metabolites and its response to UV and cadmium (Cd) exposure in S. miltiorrhiza. It was found that transgenic S. miltiorrhiza hairy roots overexpressing ScWRKY35 displayed upregulated expression of genes related to phenolic acid synthesis, resulting in increased salvianolic acid B (SAB) and rosmarinic acid (RA) contents. Conversely, tanshinone pathway gene expression decreased, leading to lower tanshinone levels. Further, overexpression of ScWRKY35 upregulated Cd transport protein HMA3 in root tissues inducing Cd sequestration. In contrast, the Cd uptake gene NRAMP1 was downregulated, reducing Cd absorption. In response to UV radiation, ScWRKY35 overexpression led to an increase in the accumulation of phenolic acid and tanshinone contents, including upregulation of genes associated with salicylic acid (SA) and jasmonic acid (JA) synthesis. Altogether, these findings highlight the role of ScWRKY35 in enhancing secondary metabolites accumulation, as well as in Cd and UV stress modulation in S. miltiorrhiza, which offers a novel insight into its phytochemistry and provides a new option for the genetic improvement of the plants.

UBC Gene Family Analysis in Salvia castanea and Roles of ScUBC2/5 Genes under Abiotic Stress.

Salvia castanea Diels, a relative of the medicinal plant Salvia miltiorrhiza Bunge, belongs to the genus Salvia and family Lamiaceae. Ubiquitin-conjugating enzyme E2 (UBC) is an important ubiquitin-binding enzyme in protein ubiquitination. This study aimed to analyze the regulatory role of UBC genes, particularly ScUBC2/5, on the growth and adaptation of S. castanea to extreme environments including cold or drought stress. We identified nine UBC genes in S. castanea and found that these genes were extremely stable and more highly expressed in the roots than other tissues. This suggested that UBC genes might play a role in promoting root adaptation to cold and dry environments. Further analysis of UBC gene expression in hairy roots under cold (4 °C) and UV stress also confirmed their importance under stress. The contents of tanshinone and salvianolic acid in hairy roots with the overexpression of ScUBC2/5 were increased compared to non-transgenic wild type, and the cold and UV resistance of hairy roots was increased compared with that of wild type. Together, these findings highlighted the role of ScUBC2/5 in enhancing secondary metabolite accumulation and regulation in response to cold and ultraviolet stress in S. castanea, providing a new perspective for genetic improvement in its phytochemistry.

Soil applied silicon and manganese combined with foliar application of 5-aminolevulinic acid mediate photosynthetic recovery in Cd-stressed Salvia miltiorrhiza by regulating Cd-transporter genes.

Salvia miltiorrhiza is an important medicinal plant that experiences significant growth and biomass losses when cultivated on cadmium (Cd) contaminated soils. High Cd accumulation in plant tissues also increases the risk of metal entry into the food chain. In this study, we proposed that Cd accumulation in S. miltiorrhiza can be restricted through plant growth regulators and nutrient management. Therefore, S. miltiorrhiza seedlings were transplanted into mixed nutrient soil for two weeks, then treated with 30 mg kg-1 CdCl2, 200 mg kg-1 Na2SiO3·9H2O, and 100 mg kg-1 MnSO4, and simultaneously sprayed with 10 mg L-1 ALA on the leaves one week later. This study showed that elevated Cd accumulation significantly reduced plant growth and biomass. This growth inhibition damaged photosynthetic machinery and impaired carbon assimilation. In contrast, 5-aminolevulinic acid (ALA) significantly promoted the biomass of S. miltiorrhiza, and the dry weight of plants treated with ALA combined with manganese (Mn)/silicon (Si) increased by 42% and 55% as compared with Cd+Mn and Cd+Si treatments. Exogenously applied ALA and Si/Mn significantly activated antioxidant enzymes and promoted the growth recovery of S. miltiorrhiza. Further, exogenous ALA also reduced the Cd concentration in S. miltiorrhiza, especially when combined with Si. Compared with the Cd+Si treatment, the Cd+Si+ALA treatment reduced the Cd concentration in roots and leaves by 59% and 60%, respectively. Gene expression analysis suggested that ALA and Si significantly up-regulated genes associated with Cd transport. Other genes related to heavy metal tolerance mechanisms are also regulated to cope with heavy metal stress. These results indicated that the combined action of ALA and Si/Mn could reduce Cd-toxicity by increasing chlorophyll content and changing oxidative stress and can also affect Cd accumulation by regulating gene expression.

Characterization of γ-glutamyltranspeptidases from dormant garlic and onion bulbs.

This study investigated the characteristics of γ-glutamyltranspeptidases (GGTs) isolated from dormant garlic (Allium sativum L.) and onion (Allium cepa L. var. agrogatum Don) bulbs. GGTs were isolated using (NH 4)2 SO 4 precipitation and hydrophobic interaction chromatography (phenyl-Sepharose column). The optimal temperature, optimal pH of extraction, and the effects of metal ions and organic compounds on the activity of GGTs were investigated. The optimal pH of the GGTs of garlic and onion was 5 and 7, respectively; the optimal temperatures were 70 and 50°C, respectively. Garlic's GGT had a major band at 53 kDa, whereas onion's GGT had two bands at 55 and 22 kDa. Cu2+, Mn2+, Fe2+, Mg2+, glucose, aspartic acid, and cysteine significantly enhanced the activity of garlic's GGT. Lysine and proline remarkably promoted the activity of onion's GGT, whereas Cu2+, glucose, and aspartic acid repress its activity. These results may deepen our understanding of allium GGTs and promote the commercial production of bioactive allium compounds.

Characterization of antioxidant activity of sulfur compounds in black garlic.

Different types of sulfur compounds, namely S-allyl-L-cysteine (SAC), S-allyl-L-cysteine sulfoxide (ACSO) and a synthetic γ-L-glutamyl-S-allyl-L-cysteine (GSAC) were extracted from black garlic and their inhibition to the advanced glycation end-products (AGEs) were investigated. Upon addition of inhibitor, the amount of produced fructosamine was determined by UV visible spectroscopy. The change of pentosidine and fluorescent AGEs during reaction was detected by fluorescence method and the change of carboxymethyl lysine (CML) was detected by high performance liquid chromatography. It was found that the inhibitory effects of SAC and ACSO are stronger to the early and mid non-fluorescent products in glycosylation reaction, and GSAC has an obvious inhibitory effect on the later reaction products. All these three inhibitors can effectively inhibit advanced glycation reaction. Although their effects on glycation products are different due to different chemical structures, they have similar inhibitory effects on fluorescent products.

Anti-hyperlipidemia of garlic by reducing the level of total cholesterol and low-density lipoprotein: A meta-analysis.

BACKGROUND: This study aimed to understand the impact of garlic on improving blood lipids using a meta-analysis. METHODS: A literature search of the PubMed, EMBASE, and Cochrane Library databases was performed using keywords such as "garlic" and "hypercholesterolemia," and the deadline "July 14 (th), 2017." After extracting relevant details, each selected literature was evaluated for quality according to the quality evaluation criteria of bias risk recommended by Cochrane Collaboration recommendations and heterogeneity tests were performed. Standardized mean difference (SMD) and 95% confidence interval (CI) were evaluated using R 3.12 software. The publication bias was assessed using Egger method. RESULTS: A total of 14 eligible papers published from 1981 to 2016 were included. The quality of the literatures was of moderate to high qualities. The values of TC (SMD = -1.26, 95% CI, -1.86 to -0.66), low-density lipoprotein (LDL) (SMD = -1.07, 95% CI, -1.67 to -0.47), and high-density lipoprotein (HDL) (SMD = 0.50, 95% CI, 0.06-0.94) after taking garlic in the experimental group and the control group have statistical significance, while there was no significant difference of TG in the 2 groups (SMD = -0.16, 95% CI, -0.87-0.55). However, the result of HDL was reversed when removed some of the literatures. No significant publication bias among the eligible studies with values of TC (P = .0625), LDL (P = .0770), HDL (P = .2293), and TG (P = .3436). CONCLUSION: Garlic can reduce the level of TC and LDL instead of HDL and TG, indicating the ability of anti-hyperlipidemia.

Changes in nutritional and bio-functional compounds and antioxidant capacity during black garlic processing.

This study aimed to explore ideal processing condition for black garlic based on the change of nutritional and active components and antioxidant capacity. Fresh garlic was processed under the condition of constant temperature (65, 75 and 85 °C) and relative humidity (70, 75, 80 and 85%) for 16 days. The sensory scores, contents of nutritional and active components, and antioxidant capacity were monitored. The sensory scores reached the maximum on the 8th day at 85% humidity and 75 °C. The contents of nutritional components were significantly affected by humidity and temperature, and 85% humidity and 75 °C were appropriate. The polyphenol content increased with increase in temperature and decrease in humidity. The reducing sugars and total sugars, total acids and 5-HMF were higher at 75 °C than at 65 and 85 °C. Reducing sugar and protein contents and sensory scores decreased on the 8th day. Maintaining the temperature of 75 °C and relative humidity of 85% for 8 days were ideal for black garlic to retain antioxidant capacity and abundant nutrients.

In vitro and in vivo antioxidant activities of polyphenol extracted from black garlic

Abstract This study investigated the in vitro and in vivo antioxidant activities of polyphenol extracted from black garlic. Black garlic polyphenol (BGP) was extracted from black garlic. The in vitro antioxidant activities of BGP were determined using DPPH·, OH and O2- radical scavenging assays. The in vivo antioxidant activities were determined by detecting the malondialdehyde (MDA) content and superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activities in mice. Results showed that, the DPPH· radical inhibition rate of 200 and 250 μg/mL BGP was equivalent with Vc (P > 0.05). With concentration of 400 and 500 μg/mL, the OH radical inhibition rate of BGP was slightly lower than Vc (P > 0.05). The O2- radical inhibition rates of 200, 400, 600, 800 and 1000 μg/mL BGP were significantly lower than Vc (P < 0.05). In the groups treated with BGP with suitable dose, the serum MDA content was significantly decreased compared with model group (P < 0.05), and the serum SOD and GSH-Px activities were significantly increased (P < 0.05). BGP has obvious DPPH· and ·OH radical scavenging activities, and can significantly decrease the serum MDA content in mice with oxidative damage, and increase the serum SOD and GSH-Px activities.

Study on the interaction of bioactive compound S-allyl cysteine from garlic with serum albumin.

Multispectroscopic techniques were used to investigate the interaction of S-allyl cysteine (SAC) from garlic with human serum albumin (HSA). UV-Vis absorption measurements prove the formation of the HSA-SAC complex. An analysis of fluorescence spectra revealed that in the presence of SAC, the quenching mechanism of HSA is considered static. The quenching rate constant Kq, KSV, and the binding constant KA were estimated. According to the Van't Hoff equation, the thermodynamic parameters enthalpy change (ΔH) and entropy change (ΔS) were calculated to be -1.00×105 J/mol and -255 J/mol/K, respectively. These indicate that hydrogen bonds and van der Waals forces are the major forces between SAC and HSA. The changes in the secondary structure of HSA, which was induced by SAC, were determined by circular dichroism spectroscopy. Energy transfer was confirmed and the distance between donor and acceptor was calculated to be 2.83 nm.

Activated Glia Increased the Level of Proinflammatory Cytokines in a Resiniferatoxin-Induced Neuropathic Pain Rat Model.

BACKGROUND AND OBJECTIVES: Administration of resiniferatoxin (RTX) can mimic the clinical symptoms of postherpetic neuralgia. However, it is unclear whether activated glia contribute to the pathogenesis of RTX-induced neuropathic pain; furthermore, the relationship between p38, N-methyl-D-aspartate receptor type 2B (NR2B) as well as proinflammatory cytokines and activated glia remains unknown. METHODS: Intraperitoneal injection of RTX was performed to induce neuropathic pain in rats. Mechanical allodynia and thermal hyperalgesia were assessed by von Frey filaments and a radiant heat stimulus, respectively. Western blot and immunofluorescence labeling examined the expression of NR2B, activated glia markers, p38, and proinflammatory cytokines in the spinal cord. We further investigated the effect of the glial inhibitors, fluorocitrate and minocycline, on nociceptive behaviors and expression of p38, NR2B, and proinflammatory cytokines. RESULTS: Resiniferatoxin leads to an increase of paw withdrawal latency to a heat stimulus and caused a mechanical allodynia within 2 weeks. The expression of tumor necrosis factor α, IL-1ß, p38, and NR2B was up-regulated in RTX-induced neuropathic pain rat model and lasted for at least 49 days. Microglia were activated at the early phase of the disease, whereas activated astrocytes were detected in the sustainment phase. Both minocycline and fluorocitrate attenuated the nociceptive behaviors and expression of related proteins. CONCLUSIONS: Activated glia participate in the pathogenesis of RTX-induced neuropathic pain and are likely to be the source of proinflammatory cytokines. Inhibition of glia contributes to an analgesic effect. These findings provide a novel strategy for the treatment of postherpetic neuralgia.

Autoxidation of unsaturated lipids in food emulsion.

Unsaturated lipids having various physiological roles are of significance in biochemistry, nutrition, medicine, and food. However, the susceptibility of lipids to oxidation is a major cause of quality deterioration in food emulsions. The reaction mechanism and factors that influence oxidation are appreciably different for emulsified lipids and bulk lipids. This article gives a brief overview of the current knowledge on autoxidation of oil-in-water food emulsions, especially those that contain unsaturated lipids, which are important in the food industry. Autoxidation of unsaturated lipids in oil-in-water emulsion is discussed, and so also their oxidation mechanism, the major factors influencing oxidation, determination measures, research status, and the problems encountered in recent years. Some effective strategies for controlling lipid oxidation in food emulsion have been presented in this review.

[Advancement on thrombolytic characteristic, function and clinical application of different fibrinolytic enzymes].

Cardio-cerebral vascular diseases endanger people's health very seriously. Thrombolytic therapy is effective in curing thrombotic diseases at present. Microorganism is an important source of thrombolytic drug. Plasminogen activators are widely used as thrombolytic drugs clinically, while they are still exists some defects. This article analyzed research and development status of kinds of thrombolytic drugs from microorganisms, and evaluated their clinical efficacy and safety, aiming at showing the direction to search new and effective thrombolytic drugs and prevent and treat thromboembolic disease.