Paeonia Suffruticosa Research Articles

Stilbenes with potent cytotoxicity from the seedcases of Paeonia suffruticosa Andrews

Stilbenes (based on the 1,2-diphenylethylene skeleton) are a class of plant polyphenols with rich structural and bioactive diversity. Twenty-six stilbenes, including five undescribed compounds (7,8-dioxy-4,3′,5′-trihydroxystilbene, trans-13′-methoxygnetin H, suffruticosol E, paestibenetrimerols A and B), were isolated from the seedcases of Paeonia suffruticosa Andrews. Their structures were elucidated by spectroscopic analyses and comparison with previously reported data. The absolute configurations of trans-13′-methoxygnetin H, suffruticosol E, paestibenetrimerols A and B were assigned from their respective electronic circular dichroism (ECD) spectra. Additionally, the structures of known compounds suffruticosols A, B and rockiol B were revised and the absolute configurations of them, and along with (+)-davidiol A, were also further determined by ECD. The isolated compounds, trans-gnetin H, cis-gnetin H and suffruticosol E, were found to have potent cytotoxicity against the DU-145 and MDA-MB-231 cell lines with IC50 values of 4.89–8.61 μM. The preliminary antitumor structure-activity relationship of these stilbenes is discussed as well.

Characteristics of PoVIN3, a Key Gene of Vernalization Pathway, Affects Flowering Time.

The tree peony (Paeonia section Moutan DC.) is the candidate flower in China, with abundant germplasm resources and high ornamental value. However, the short and concentrated flowering period severely restricted the improvement of the economic value of tree peonies. Based on the full-length transcriptome database of tree peonies, the PoVIN3 (GenBank ID: OP341879), involved in the flowering regulation of tree peonies were identified and cloned for the first time. The PoVIN3 was also characterized by bioinformatics methods, quantitative real-time PCR (qRT-PCR), and the establishment of a transgenic system. The expression levels of PoVIN3 in seven different petals developmental stages were the highest at the initial flowering stage of the variant cultivar of Paeonia ostii 'Fengdan,' the initial decay stage of the normal flowering Paeonia ostii 'Fengdan,' and the half opening stage of the late flowering Paeonia suffruticosa 'Lianhe.' Tissue-specific expression analysis showed that the relative expression levels of PoVIN3 were the highest in sepals of both normal flowering Paeonia ostii 'Fengdan' and the late flowering Paeonia suffruticosa 'Lianhe,' and the highest expression was in stamens of early flowering mutant Paeonia ostii 'Fengdan.' In addition, the flowering time of pCAMBIA2300-PoVIN3 transgenic plants was significantly earlier than that of the wild-type, indicating that PoVIN3 could promote plant flowering. The results provide a theoretical basis for exploring the role of PoVIN3 in the regulation of flowering in tree peonies.

Chemical fingerprinting and multivariate analysis of Paeonia ostii leaves based on HPLC-DAD and UPLC-ESI-Q/TOF-MS/MS

Paeonia suffruticosa Andr. (Tree peony) is a well-known traditional Chinese medicine, which has been widely cultivated for thousands of years and its root bark is used in many prescriptions. Oil peony (Paeonia ostii and Paeonia rockii) has been planted in large areas since tree peony seed oil was approved as a new food resource by the National Health Commission (Ministry of Health) of P.R. China in 2011. However, tree peony leaves were often regarded as waste resources during cultivation and there were few comparative studies on the chemical constituents of tree peony leaves from different origins. In this study, thirty-five P. ostii leaves from four provinces in China were taken as the research objects. The methods of high-performance liquid chromatography (HPLC-DAD) fingerprinting combined with multivariate analysis, including similarity analysis (SA), principal component analysis (PCA), orthogonal partial least square method-discriminant analysis (OPLS-DA) and hierarchical cluster analysis (HCA), were used to evaluate the difference of chemical composition between thirty-five samples. UPLC-ESI-Q/TOF-MS/MS and HPLC-DAD were used for qualitative and quantitative analysis of characteristic components. The results showed that seventeen common characteristic peaks were screened from HPLC fingerprints and the similarity of thirty-five samples was between 0.864 and 0.999. Thirty-five samples from different origin were divided into four groups by OPLS-DA and PCA, but not HCA. Meanwhile, ten compounds were found to be the most closely related marker components for quality evaluation. In addition, 56 characteristic chemical components were preliminarily identified from P. ostii leaves by UPLC-ESI-Q/TOF-MS/MS technology. Finally, the quantitative analysis results showed that there were some differences in the chemical contents of seven characteristic components in thirty-five P. ostii leaves. This study provided a reasonable method for the discrimination and quality evaluation of P. ostii leaves from different producing areas, and laid a foundation for the comprehensive utilization and development of P. ostii resources.

Identification and Pathogenicity of Pestalotiod Fungi Associated with Woody Oil Plants in Sichuan Province, China.

Pestalotiod fungi are associated with a wide variety of plants worldwide and occur as endophytes, pathogens, and saprobes. The present study provides an updated phylogeny for genera Neopestalotiopsis, Pestalotiopsis, and Seiridium using fresh collections from woody oil plants (Camellia oleifera, Olea europaea, Paeonia suffruticosa, Sapium sebiferum, and Vernicia fordii) in Sichuan Province, China. We coupled morphology and combined sequence data analyses of ITS, tub2, and tef1-α for Neopestalotiopsis and Pestalotiopsis, with ITS, LSU, tub2, tef1-α, and rpb2 for Seiridium. Three novel species of Neopestalotiopsis (N. mianyangensis, N. paeonia-suffruticosa, N. terricola) and three of Seiridium (S. guangyuanum, S. vernicola, S. oleae), were found. Three other species, Pestalotiopsis kenyana, Seiridium ceratosporum, and S. rosarum were identified and reported as new records. All isolated species are fully described and illustrated. Additionally, the sexual morph of Pestalotiopsis kenyana is described for the first time. Pathogenicity tests revealed that Neopestalotiopsis mianyangensis, N. paeonia-suffruticosa, N. terricola, Pestalotiopsis kenyana, Seiridium guangyuanum, S. vernicola, and S. oleae are pathogenic on detached olive leaves.

Suffruticosol B Is an Osteogenic Inducer through Osteoblast Differentiation, Autophagy, Adhesion, and Migration.

Suffruticosol B (Suf-B) is a stilbene found in Paeonia suffruticosa ANDR., which has been traditionally used in medicine. Stilbenes and their derivatives possess various pharmacological effects, such as anticancer, anti-inflammatory, and anti-osteoporotic activities. This study aimed to explore the bone-forming activities and mechanisms of Suf-B in pre-osteoblasts. Herein, >99.9% pure Suf-B was isolated from P. suffruticosa methanolic extracts. High concentrations of Suf-B were cytotoxic, whereas low concentrations did not affect cytotoxicity in pre-osteoblasts. Under zero levels of cytotoxicity, Suf-B exhibited bone-forming abilities by enhancing alkaline phosphatase enzyme activities, bone matrix calcification, and expression levels with non-collagenous proteins. Suf-B induces intracellular signal transduction, leading to nuclear RUNX2 expression. Suf-B-stimulated differentiation showed increases in autophagy proteins and autophagosomes, as well as enhancement of osteoblast adhesion and transmigration on the ECM. These results indicate that Suf-B has osteogenic qualities related to differentiation, autophagy, adhesion, and migration. This also suggests that Suf-B could have a therapeutic effect as a phytomedicine in skeletal disorders.

Integrative analysis of transcriptomic and volatile compound profiles sheds new insights into the terpenoid biosynthesis in tree peony

Tree peony (Paeonia suffruticosa Andr.), a traditional Chinese ornamental plant, is famous for its large, colorful, and fragrant flowers. Studies on tree peony have mainly focused on the improvement of flower color and the regulation of the flowering stage, but the molecular mechanisms controlling the formation of floral fragrance has yet to be understood. In this study, the aromatic tree peony variety ‘High Noon’ was used to determine whether volatile compounds (volatiles) in flowers were synthesized at the balloon, initial opening, half opening, or full opening stages. Transcriptomic sequencing was conducted on petals and stamens collected at the balloon and full opening stages. The relative content of linalool peaked during the full opening stage of flowering, and linalool was the main component of flower fragrance. As flowering progressed, the relative content of monoterpenes gradually increased and the relative content of sesquiterpenes gradually decreased. Transcriptome data revealed 23695 differentially expressed genes (DEGs), 142 of which were related to terpene accumulation. Transcriptome analysis and correlation analysis between floral volatiles and gene expressions showed that, 1-deoxy-d-xylulose 5- phosphate synthase (DXS), 1-deoxy-d-xylulose 5-phosphate reductoisomerase (DXR), acetyl-CoA acetyltransferase (AACT), geranyl pyrophosphate synthase (GPPS), and terpene synthase (TPS) may be major regulators controlling floral terpene biosynthesis in ‘High Noon’. And PsGPPS may regulate the expression of GPPS, and then participate in the synthesis of terpenes. This study provides a possible molecular mechanism for terpene formation in tree peony ‘High Noon’ flowers as well as a theoretical basis upon which to study tree peony flowers.

Rapid Identification of Paeoniae Radix and Moutan Radicis Cortex Using a SCAR Marker-Based Conventional PCR Assay.

Paeoniae Radix is a herbal medicine prepared from the dried roots of Paeonia lactiflora, P. anomala subsp. veitchii, and P. japonica. Although the herbal medicines prepared from these species are morphologically similar, they have different pharmacological effects depending on how they are processed. In addition, P. japonica is more expensive than other Paeonia spp. in the Korean herbal market. Although there is a clear difference between the Korean and Chinese pharmacopeias of Paeoniae Radix, the processed roots of P. lactiflora and P. anomala subsp. veitchii are commonly used indiscriminately in the herbal market. Moreover, Paeonia suffruticosa, an allied genus of P. lactiflora, is prescribed as Moutan Radicis Cortex. Therefore, accurate taxonomic identification of plant species is vital for quality assurance. A genetic assay is a reliable tool for accurately discriminating species in processed herbal medicines. To develop a genetic assay for the identification of four Paeonia species (P. lactiflora, P. anomala subsp. veitchii, P. japonica, and P. suffruticosa), we analyzed the sequences of two DNA barcoding regions, internal transcribed spacer and rbcL. A conventional PCR assay was established in this study for simple and rapid species identification using sequence characterized amplified region (SCAR) markers based on arbitrary nucleotide-containing primers. This assay was verified to be species specific and highly sensitive and could be applied to Paeonia species identification at an affordable rate.

Preliminary Research on the Effects of Different Substrates on the Metabolome of Potted Peonies

Peony (Paeonia suffruticosa Andr.) is a traditional ornamental flower in China. Peony potted culture has become mainstream. The development of agricultural and forestry waste instead of a nonrenewable peat matrix as the substrate is an important direction for the future industry of potted peonies. However, the mechanism of the effects of different substrates on the growth and development of peonies remains unclear. In this experiment, “Luhehong”, a peony variety from Heze, Shandong Province, was selected as the main experimental material. The response mechanism of peony roots to different substrate proportions was analyzed using untargeted metabolomics of potted peonies. Results showed that the potential for pot growth of planted peonies using the mixed-straw mixing matrix (Matrix K) is better than that of the rice husk matrix (Matrix G) and the control matrix. Moreover, the comparative analysis of the metabolic groups indicates that the number of peony root secretions in the three matrices has a remarkable difference, possibly related to the nutrition supply levels of the peony roots, in the different matrices of adaptive response and adjustment. Furthermore, (2r)-3-hydroxyisovaleroylcarnitine, trigonelline, 2-acetylresorcinol, and dehydroascorbic acid (oxidized vitamin C)—four metabolites with the most substantial intergroup differences—are considered key metabolites. The metabolic pathway analysis of propanoate, and ascorbate and aldarate metabolism, and the two pathways related to an environmental response, namely, ABC transporters and phosphatidylinositol signaling system, indicated that these pathways were remarkably enriched. Both may be the key way to affect the growth and development of potted peonies. This study can provide an important reference for the rational allocation of the substrate of agricultural and forestry waste in potted peony cultivation and application.

Air Layering Improves Rooting in Tree Peony Cultivars from the Jiangnan Group

Tree peony (Paeonia suffruticosa Andr.), a unique traditional flower in China, is famous for its ornamental value, medical use, and edible oil production. Traditional propagation methods, such as sowing, dividing, and grafting do not allow the large-scale production of selected peony varieties. Therefore, the objective of our study is to evaluate an air-layering technique on the rooting success of three tree peony cultivars (‘Baoqing Hong’, ‘Quehao’, and ‘Xishi’). The experiments were established through consideration of the influence of the time of the year the rooting was performed (mid-May, mid-June, or mid-July) and the growth regulators (1-naphthaleneacetic acid-NAA and indole-3-butyric acid-IBA) applied at different concentrations (1000 mg/L, 1500 mg/L, 2000 mg/L). The results showed that the rooting rate was the highest when the air-layering time occurred in mid-June, and the rooting rate of ‘Quehao’ was found to be the most significant, reaching 100%. The rooting percentages of 82.86% and 77.14% were obtained for ‘Baoqing Hong’ and ‘Xishi’, respectively. The growth regulators affected the rooting performance of the three cultivars differently. The rooting parameters of ‘Quehao’ were negatively correlated with the concentration of NAA but positively correlated with IBA, whereas the ‘Baoqing Hong’ and ‘Xishi’ cultivars showed no dose dependence for the supplied growth regulators. Root number, root tip number, and maximum root length in ‘Quehao’ were higher than those of the other two cultivars. The conclusion of our study is that the air-layering technique is a suitable method for achieving satisfactory propagation of selected tree peony cultivars.

Preparation of chitosan/peony (Paeonia suffruticosa Andr.) leaf extract composite film and its application in sustainable active food packaging

Considering the problem of food perishability and the environmental pollution of plastic packaging, the natural active packaging prepared by incorporating plant leaf (e.g. peony leaf) extract into biodegradable food packaging materials may be a key to addressing these issues. In the study, a novel green bioactive composite film (CS-PLE) with antioxidant and antibacterial propertiese was developed by blending peony leaf extract (PLE) into chitosan (CS) film through the solution casting method. The physical properties and biological activities of a series of films, including CS film and CS-PLEs incorporated with different concentrations of PLE, were studied by appropriate experimental protocols. The results demonstrated that addition of PLE improved thermal stability, the barrier performance of UV-A and total phenolic content of CS film, and the antioxidant activities of CS-PLEs increased by 60.6 % ~ 73.2 % with the appending proportion of PLE from 0.1 wt% ~ 0.7 wt% compared to CS film. Furthermore, CS-PLEs had better inhibiting effect on bacteria. Notably, CS-PLE (0.7 wt%) wrapping apples showed the least degree of browning among all the groups. Hence, the study of CS-PLEs can provide a new strategy for the preparation of food packaging materials that are green alternatives and reduce environmental pollution.

Grain oil quality formation and metabolism-related genes difference expression of Paeonia suffruticosa cv. 'Fengdan' grown at different altitudes.

We measured the morphological index, nutritional composition and the expression analysis of key genes during grain development of Paeonia suffruticosa cv. 'Fengdan' grown at altitudes of 100, 650 and 1010 m in Luo-yang. The aim of this study was to examine differences in grain yield traits and the transformation of soluble sugar, starch, soluble protein and fatty acid contents, as well as the related enzyme activity and differential expression of key genes in oil metabolism. The results showed that grain yield traits increased with altitudes and that the growth period of grain at the higher altitudes was longer than that at low and mid altitudes. The soluble sugar and starch in mature grains increased with altitudes, while soluble protein and crude fat did not change. During grain development, the activities of sucrose synthase (SS) and sucrose phosphate synthase (SPS) first decreased and then increased, with the lowest occurred at 90 d after flowering. The activities of pyruvate dehydrogenase (PDH), glutamic-pyruvic transaminase (GPT) and glutamic-oxalacetic transaminease (GOT) increased rapidly during 50-90 d after flowering and peaked at 90 d. The relative expression of acetyl-CoA carboxylase (ACCase) and stearoyl-ACP desaturase (SAD) peaked at 50 d after flowering, and ω-6 fatty acid desaturase 2 (FAD2) peaked at 90 d, in oil tree peony grain at different altitudes. There was a negative correlation of soluble sugar and starch with the accumulation of soluble protein and crude fat. SPS activity was positively correlated with the contents of soluble sugar and starch, and negatively correlated with the contents of soluble protein and crude fat during grain development. Activities of GPT and GOT were negatively associated with the content of soluble sugar and the content of starch, and had a highly significant positive correlation with the contents of soluble protein and crude fat. Activity of PDH was positively correlated with the content of soluble proteins and activities of GPT and GOT, and negatively correlated with the contents of soluble sugar and starch. It suggested that nutrient accumulation in the process of grain development of tree peony was transformed from sugar to crude fat and protein, and that metabolic enzymes, such as SPS, PDH, GPT and GOT, played an important role in this process. Palmitate acid, stearic acid and linoleic acid were negatively correlated with the relative increment of α-linolenic acid, indicating that fatty acid desaturation process in the grain development of tree peony was towards the direction of α-linolenic acid synthesis. The relative expression of ACCase, SAD, and FAD2 was positively correlated with the relative increment of α-linolenic acid accumulation, which played an important role in α-linolenic acid synthesis. The oil quality of tree peony grain was relatively stable at different altitudes, but grain production increased with altitude. Planting oil tree peony at mid to high altitudes could be an important strategy for the efficient use of marginal land in Luoyang.

Complete defoliation is essential for tree peony reflowering in autumn to trigger sucrose metabolism and adjust source-sink balance in buds and leaves

Tree peony (Paeonia suffruticosa Andr.) has a short and concentrated natural fluorescence, and forcing culture is frequently used for its annual production. We previously found that defoliation is critical for tree peony to reflower in the autumn. In this study, we investigated how defoliation induced this process in more detail. We found that partial defoliation with only three top leaves remaining failed to trigger bud development or reflowering. Furthermore, complete defoliation, but not partial defoliation, decreased the contents of sucrose in buds, as well as increasing the contents of glucose and fructose in buds and leaves. Therefore, sucrose metabolism, particularly its hydrolysis, may function as a switch for tree peony reflowering, which is triggered by complete defoliation. Since the complete and partial defoliation treatments produced similar physical damage to plants, the induction of reflowering was unlikely to be related to a stress signal. In addition, the net photosynthetic rates of mature and newly grown young leaves in partial and complete defoliation groups, respectively, were lower than those of the control. Thus, photosynthesis itself may not be involved in this process. At the transcriptional level, defoliation treatment, but not partial defoliation, significantly induced PsSUT1 and PsSWEET12 expression. This was accompanied by the reduction of PsSWEET7 at 24 d in buds, which could facilitate the transfer of sucrose from the leaves to buds. PsCIN3, PsVIN1 and PsVIN2 were also substantially induced by defoliation treatment rather than partial defoliation, which should be mainly involved in sucrose metabolism. In short, complete defoliation is essential for tree peony reflowering in the autumn by triggering sucrose metabolism, as well as rebalancing the source-sink relationship in buds and leaves.

Development and Characterization of Microsatellite Markers Based on the Chloroplast Genome of Tree Peony.

Tree peony (Paeonia suffruticosa Andr.) is a famous ornamental and medicinal flowering species. However, few high-efficiency chloroplast microsatellite markers have been developed for it to be employed in taxonomic identifications and evaluation of germplasm resources to date. In the present study, a total of 139 cpSSR loci were identified across eleven tree peony plastomes. Dinucleotide repeat SSRs (97.12%) were most abundantly repeated for the AT motif (58.27%), followed by the TA motif (30.94%) and the TC motif (7.91%). Twenty-one primer pairs were developed, and amplification tests were conducted for nine tree peony individuals. Furthermore, 19 cpSSR markers were amplified on 60 tree peony accessions by a capillary electrophoresis test. Of 19 cpSSR markers, 12 showed polymorphism with different alleles ranging from 1.333 to 3.000. The Shannon’s information index and polymorphism information content values ranged from 0.038 to 0.887 (mean 0.432) and 0.032 to 0.589 (mean 0.268), respectively. The diversity levels for twelve loci ranged from 0.016 (at loci cpSSR-8 and cpSSR-26) to 0.543 (at locus cpSSR-15), averaging 0.268 for all loci. A total of 14 haplotypes (23.33%) were detected in the three populations. The haplotypic richness ranged from 0.949 to 1.751, with a mean of 1.233 per population. The genetic relationship suggested by the neighbor-joining-based dendrogram divided the genotypes into two clusters. The Jiangnan population was allotted to Cluster II, and the other two populations were distributed into both branches. These newly developed cpSSRs can be utilized for future breeding programs, population genetics investigations, unraveling the genetic relationships between related species, and germplasm management.

Antioxidant Activity, Functional Properties, and Cytoprotective Effects on HepG2 Cells of Tree Peony (Paeonia suffruticosa Andr.) Seed Protein Hydrolysate as Influenced by Molecular Weights Fractionation.

In recent years, plant protein hydrolysates have gained increased attention due to their superior antioxidant activity and potential to prevent several chronic diseases associated with oxidative stress. This study aimed to investigate the antioxidant activity, functional properties, and cytoprotective effects of the tree peony seed protein hydrolysate (TPSPH) with different molecular weights (MWs). The antioxidant activities were evaluated by DPPH, hydroxyl radicals scavenging, Fe2+ chelating, and inhibition of the β-carotene oxidation abilities. The protective effects and mechanism against oxidative stress were determined using H2O2-stressed HepG2 cells. MW > 30 kDa of TPSPH showed the highest radical scavenging (DPPH IC50 = 0.04, hydroxyl IC50 = 0.89 mg/mL) and inhibition of β-carotene oxidation (80.07% at 2.0 mg/mL) activity. Moreover, MW > 30 kDa possessed high hydrophobicity, emulsifying capacity, and abundant antioxidant amino acids (28.22% of hydrophobic amino acids and 8.3% of aromatic amino acids). MW 5–10 kDa exhibited more effective protection against H2O2-induced HepG2 cells, by reducing reactive oxygen species (ROS), malonaldehyde (MDA), lactate dehydrogenase (LDH), and activating antioxidant enzymes (superoxide dismutase and catalase). These results indicated the potential application of TPSPH as an antioxidant in food and functional foods.

Deciphering the microbial diversity associated with healthy and wilted Paeonia suffruticosa rhizosphere soil.

Plant health is closely related to the soil, where microorganisms play a critical and unique role. For instance, Paeonia suffruticosa is an emerging woody oil crop in China with attractive development and utilization prospects. However, black root rot causes wilting of the aboveground plant parts, which significantly affected its seed yield and quality. Studies found that soil microorganisms are critical in maintaining plant health, but how changes in the soil microbial communities affect the healthy and diseased oil peony is unclear. Therefore, our present study used high throughput sequencing and BIOLOG to analyze the rhizosphere soil microbial communities of healthy and diseased oil peonies. Our results revealed that the physical and chemical properties of the soil of the diseased plants had changed, with the ability to metabolize the carbon source being enhanced. Moreover, our research highlighted that the oil peony-infecting fungal pathogenic genus (Fusarium, Cylindrocarpon, and Neocosmospora) was closely associated with oil peony yield reduction and disease aggravation. Further network analysis demonstrated that the bacterial and fungal networks of the diseased plants were more complex than those of the healthy plants. Finally, the inter-kingdom network among the diseased plants further indicated that the lesions destroyed the network and increased the intraspecific correlation between the fungal groups.

Integrating network pharmacology analysis and pharmacodynamic evaluation for exploring the active components and molecular mechanism of moutan seed coat extract to improve cognitive impairment.

Paeonia suffruticosa (Moutan) is a traditional medicinal plant in China. Its seed coat is rich in resveratrol oligomer, especially suffruticosol B (SB). Previous studies had shown that the seed coat extracts of Paeonia suffruticosa (PSCE) had good cholinesterase inhibitory activity and neuroprotective effect, but the effective dose range was unknown, and the pharmacodynamic components and molecular mechanism of PSCE had not been discussed. The current study aimed to screen the pharmacodynamic components in PSCE and investigate the improvement effect of PSCE and the selected SB on scopolamine-induced cognitive dysfunction in mice and its mechanism. The results of high-throughput sequencing and bioinformatics analysis showed that suffruticosol B (SB) and trans-gnetin H (GH) might be the main active components of PSCE; PSCE might improve cognitive dysfunction through p53, HIF-1, MAPK, and PI3K-Akt signaling pathways, while SB and GH might improve cognitive dysfunction through HIF-1 signaling pathway. SB and GH had good molecular docking activity with the target of HIF-1 signaling pathway. The pharmacodynamic activities of PSCE and SB were further verified by behavioral experiments. PSCE and SB could improve the recognition ability of familiar and new objects and shorten the escape latency in the Morris Water Maze test (PSCE 120 mg∙kg-1, p < 0.05; SB 60 mg∙kg-1, p < 0.01); PSCE and SB could increase Ach and GSH levels, enhance the activities of ChAT, SOD and CAT, decrease the levels of IL-1β, IL-6, and TNF-α, and decrease the activity of AChE. In conclusion, the results indicated that PSCE might exert pharmacodynamic activity through multiple components, targets, and pathways, and SB and GH might be the main active components of PSCE. PSCE and SB might improve cognitive dysfunction by regulating cholinergic, antioxidant, and anti-inflammatory effects. These results indicated that PSCE and SB might be potential anti-AD drug candidates, providing a scientific basis for the development and utilization of Moutan bark.

Physiological and transcriptional responses to heat stress and functional analyses of PsHSPs in tree peony (Paeonia suffruticosa).

Tree peony (Paeonia suffruticosa) is a traditional Chinese flower that is not resistant to high temperatures, and the frequent sunburn during summer limits its normal growth. The lack of understanding of the molecular mechanisms in tree peony has greatly restricted the improvement of novel heat-tolerant varieties. Therefore, we treated tree peony cultivar “Yuhong” (P. suffruticosa “Yuhong”) at normal (25°C) and high temperatures (40°C) and sequenced the transcriptomes, to investigate the molecular responsive mechanisms to heat stress. By comparing the transcriptomes, a total of 7,673 differentially expressed genes (DEGs) were detected comprising 4,220 upregulated and 3,453 downregulated genes. Functional annotation showed that the DEGs were mainly related to the metabolic process, cells and binding, carbon metabolism, and endoplasmic reticulum protein processing. qRT-PCR revealed that three sHSP genes (PsHSP17.8, PsHSP21, and PsHSP27.4) were upregulated in the response of tree peony to heat stress. Tissue quantification of the transgenic lines (Arabidopsis thaliana) showed that all three genes were most highly expressed in the leaves. The survival rates of transgenic lines (PsHSP17.8, PsHSP21, and PsHSP27.4) restored to normal growth after high-temperature treatment were 43, 36, and 31%, respectively. In addition, the activity of superoxide dismutase, accumulation of free proline, and chlorophyll level was higher than those of the wild-type lines, while the malondialdehyde content and conductivity were lower, and the membrane lipid peroxidation reaction of the wild-type plant was more intense. Our research found several processes and pathways related to heat resistance in tree peony including metabolic process, single-organism process, phenylpropane biosynthesis pathway, and endoplasmic reticulum protein synthesis pathway. PsHSP17.8, PsHSP21, and PsHSP27.4 improved heat tolerance by increasing SOD activity and proline content. These findings can provide genetic resources for understanding the heat-resistance response of tree peony and benefit future germplasm innovation.

Isolation and identification of novel angiotensin I-converting enzyme (ACE) inhibitory peptides from Pony Seed and evaluation of the inhibitory mechanisms

In this work, four novel angiotensin I-converting enzyme inhibitory (ACEI) peptides were isolated and identified from peony (Paeonia suffruticosa Andr.) seed hydrolysate (PSH) by consecutive chromatography and UPLC-ESI-QTOF-MS/MS. The IC50 values of HWS (0.640 ±0.016 mg/mL) and VLSGF (0.328 ± 0.040 mg/mL) were significantly (p < 0.05) lower than those of the other two peptides. The Lineweaver-Burk plots showed that VLSGF is competitive inhibition against ACE and HWS acted as a mixed-type inhibitor. Meanwhile, the results of molecular docking showed hydrogen interactions between VLSGF/HWS and ACE. In human endothelial cells, the pretreatments of VLSGF and HWS could increase the production of nitric oxide (NO) and the expression of endothelial nitric oxide synthase (eNOS). In addition, they also decreased the reactive oxygen species (ROS) production. These results indicated that it may be possible to use peony seed protein hydrolysate to create ingredients for functional foods.

640 Exploring the synergic effects of a plant and a peptide on hair follicle pigmentation

Canities is an age-linked loss of bulb melanocyte and hair follicle (HF) pigmentation. Canities can significantly alter the physical appearance of a person and negatively impact their well-being, which drives the search and demand for new anti-greying agents. αMSH is pro-pigmentary hormone of the melanocortin family, which stimulates melanogenesis through its target receptor, MC1. GreyverseTM (Gv) is a biomimetic synthetic peptide derived to mimic αMSH and is an established promoter of HF melanogenesis with redox properties. Preliminary studies revealed that cotreatment of Gv and Paeonia Suffruticosa root bark extract (PS) synergically induces mitochondrial E3 ligase (MITOL) and melanogenesis in human melanocytes in vitro. Thus, this study investigated the additive effects of PS and Gv on hair pigmentation ex vivo. HFs were cultured in serum-free medium, supplemented with PS and/or Gv for 4 days, then assessed for protein expression and enzymatic changes associated to melanogenesis. Quantitative immune-histomorphometry revealed that PS and Gv cotreatment led to an additive increase in melanin content (p<0.05) that derived from a synergic increase in Gp100 expression (p<0.05), an increase in tyrosinase activity, and αMSH and TYRP expression within the hair follicle pigmentary unit. Taken together, PS and Gv showed synergic effects in promoting hair pigmentation, primarily through melanosome maturation. Thus, this combinational treatment deserves further pre-clinical and clinical exploration for the reversal of hair greying.

Suffruticosol A elevates osteoblast differentiation targeting BMP2-Smad/1/5/8-RUNX2 in pre-osteoblasts.

The Paeonia suffruticosa ANDR. (P. suffruticosa) is commonly used in traditional medicine for various purposes. Suffruticosol A (Suf-A), isolated from P. suffruticosa, is a beneficial compound with antibiofilm, antivirulence, and anti-inflammatory properties. The aim of the present study was to investigate the biological effects of Suf-A on osteogenic processes in pre-osteoblasts. It was determined here in that Suf-A (>98.02%), isolated from P. suffruticosa, showed no cytotoxicity at 0.1-30 μM; however, it induced cytotoxicity at 50-100 μM in pre-osteoblasts. Suf-A increased osteogenic alkaline phosphatase activity and expression levels of noncollagenous proteins. Adhesion and trans-migration on the extracellular matrix were potentiated by Suf-A, but not by wound-healing migration. Suf-A did not affect autophagy or necroptosis during osteoblast differentiation. We found that Suf-A increased runt-related transcription factor 2 (RUNX2) levels and mineralized matrix formation. RUNX2 expression was mediated by Suf-A-induced BMP2-Smad1/5/8 and mitogen-activated protein kinase signaling, as demonstrated by Noggin, a BMP2 inhibitor. These results suggest that Suf-A is a potential natural osteogenic compound.