Copaiba oil minimizes inflammation and promotes parenchyma re-epithelization in acute allergic asthma model induced by ovalbumin in BALB/c mice.

Introduction: Asthma is a condition of airflow limitation, common throughout the world, with high mortality rates, especially as it still faces some obstacles in its management. As it constitutes a public health challenge, this study aimed to investigate the effect of copaiba oil (e.g., Copaifera langsdorffii), as a treatment resource, at doses of 50 and 100 mg/kg on certain mediators of acute lung inflammation (IL-33, GATA3, FOXP3, STAT3, and TBET) and early mechanisms of lung remodeling (degradation of elastic fiber tissues, collagen deposition, and goblet cell hyperplasia). Methods: Using an ovalbumin-induced acute allergic asthma model in BALB/c mice, we analyzed the inflammatory mediators through immunohistochemistry and the mechanisms of lung remodeling through histopathology, employing orcein, Masson's trichrome, and periodic acid-Schiff staining. Results: Copaiba oil treatment (CO) reduced IL-33 and increased FOXP3 by stimulating the FOXP3/GATA3 and FOXP3/STAT3 pathways. Additionally, it upregulated TBET, suggesting an additional role in controlling GATA3 activity. In the respiratory epithelium, CO decreased the fragmentation of elastic fibers while increasing the deposition of collagen fibers, favoring epithelial restructuring. Simultaneously, CO reduced goblet cell hyperplasia. Discussion: Although additional research is warranted, the demonstrated anti-inflammatory and re-epithelializing action makes CO a viable option in exploring new treatments for acute allergic asthma.

Baccharis dracunculifolia DC Consumption Improves Nociceptive and Depressive-like Behavior in Rats with Experimental Osteoarthritis.

Osteoarthritis (OA) persistently activates nociceptors, leading to chronic pain, which is often accompanied by the comorbid development of emotional impairments (anxiety and depression), an effect associated with microgliosis. Baccharis dracunculifolia DC (Asteraceae), a Brazilian edible plant, is an important source of active compounds with anti-inflammatory abilities. Thus, we evaluated its ability to reverse OA-induced nociceptive and emotional-like impairments in osteoarthritic ovariectomized female rats using the kaolin/carrageenan (K/C) model. Four weeks after OA induction, mechanical hyperalgesia was confirmed, and the treatment started. Control animals (SHAMs) were treated with phosphate-buffered saline (PBS), while arthritic animals (ARTHs) either received PBS or B. dracunculifolia 50 mg/kg (Bd50) and 100 mg/kg (Bd100), via gavage, daily for five weeks. At the end of the treatment, anxiety-like behavior was assessed using the Open Field Test (OFT), anhedonia was assessed using the Sucrose Preference Test (SPT), and learned helplessness was assessed using the Forced Swimming Test (FST). After occision, microglia were stained with IBA-1 and quantified in brain sections of target areas (prefrontal cortex, amygdala, and periaqueductal grey matter). Treatment with B. dracunculifolia extract reversed OA-induced mechanical hyperalgesia and partly improved depressive-like behavior in OA animals' concomitant to a decrease in the number of M1 microglia. Our findings suggest that B. dracunculifolia extracts can potentially be used in the food industry and for the development of nutraceuticals and functional foods.

Chimerism of avian IgY-scFv and truncated IgG-Fc: A novel strategy in cross-species antibody generation and enhancement.

Chicken single-chain fragment variable (IgY-scFv) is a functional fragment and an emerging development in genetically engineered antibodies with a wide range of biomedical applications. However, scFvs have considerably shorter serum half-life due to the absence of antibody Fc region compared with the full-length antibody, and usually requires continuous intravenous administration for efficacy. A promising approach to overcome this limitation is to fuse scFv with immunoglobulin G (IgG) Fc region, for better recognition and mediation by the neonatal Fc receptor (FcRn) in the host. In this study, engineered mammalian ΔFc domains (CH2, CH3, and intact Fc region) were fused with anti-canine parvovirus-like particles avian IgY-scFv to produce chimeric antibodies and expressed in the HEK293 cell expression system. The obtained scFv-CH2, scFv-CH3, and scFv-Fc can bind with antigen specifically and dose-dependently. Surface plasmon resonance investigation confirmed that scFv-CH2, scFv-CH3, and scFv-Fc had different degrees of binding to FcRn, with scFv-Fc showing the highest affinity. scFv-Fc had a significantly longer half-life in mice compared with the unfused scFv. The identified ΔFcs are promising for the development of engineered Fc-based therapeutic antibodies and proteins with longer half-lives. The avian IgY-scFv-mammalian IgG Fc region opens up new avenues for antibody engineering, and it is a novel strategy to enhance the rapid development and screening of functional antibodies in veterinary and human medicine.

Dendrobium nobile Lindl ameliorates learning and memory deficits in scopolamine-treated mice.

ETHNOPHARMACOLOGICAL RELEVANCE: Dendrobium nobile Lindl (DNL), a valued time-honored herb, possesses immune-boosting and age-delaying properties, has been widely used to treat hyperglycemia and neurological diseases, and is probably a potential drug for improving learning and memory. Scopolamine (Scop), an antagonist for muscarinic receptors, potentially impairing intelligence and memory. AIM OF THE STUDY: This investigation aimed to assess the efficacy of DNL in alleviating scopolamine-induced cognitive deficits in mice and its mechanisms. MATERIALS AND METHODS: We utilized the open-field test, novel object recognition test (NOR), and Morris water maze test (MWM) to assess the potential of DNL in ameliorating learning and memory dysfunction caused by scopolamine in mice. Enzyme-linked immunosorbent assay (ELISA) was employed to measure Choline acetyltransferase (ChAT) content and Acetylcholinesterase (AChE) activities in the brain, and oxidative stress-related factors in the serum, including Malondialdehyde (MDA), Superoxide dismutase (SOD), and glutathione (GSH) content. RESULTS: Scopolamine injection significantly reduced the discrimination index of mice in the NOR test and impaired their performance in the MWM test, as demonstrated by longer escape latency, fewer target crossings, and less time spent in the target quadrant in the MWM. After 25 days of administration, DNL increased the discrimination index of the scopolamine-treated mice in the NOR test. DNL reduced the escape latency in the MWM test in the model mice. DNL increased the target crossing number and the percentage of time spent in the target quadrant in the MWM test. ELISA experiments indicated that DNL decreased the AChE activities, increased the ChAT activities, and modulated oxidative stress makers (GSH, SOD, and MDA) in scopolamine-induced mice. CONCLUSIONS: DNL may improve the learning and memory in mice treated with scopolamine, possibly by modulating oxidative stress and impaired cholinergic function.

Genista tridentata Phytochemical Characterization and Biological Activities: A Systematic Review.

Genista tridentata (L.) Willk., known as "prickled broom", is a Leguminosae (Fabaceae) species native to the Iberian Peninsula, Morocco, Algeria, and Tunisia. It is used in folk medicine as an anti-inflammatory, for gastrointestinal and respiratory disorders, rheumatism, and headaches, to lower blood pressure, against hypercholesterolemia and hyperglycemia. This study aimed to systematically review the literature on the bioactivities and phytochemical profile of Genista tridentata to understand its pharmacological potential. For this, four electronic databases (PubMed, GoogleScholar, Repositórios Cientificos de Acesso Aberto de Portugal (RCCAP), and ScienceDirect) were searched from inception up to 31 December 2022. From a total of 264 potentially eligible studies considered for screening, 34 papers were considered eligible for this systematic review. The sampling included 71 extracts, collected mainly in Portugal. Genista tridentata extracts present a high level of flavonoids and phenolic compounds. The flowers and aerial parts of the plant were the most studied, and aqueous extracts were the most used. The results predict a high potential for the application of Genista tridentata as a new source of natural antioxidants and preservatives for the food industry with subsequent health benefits, such as the production of nutraceuticals. Moreover, the results indicate that the plant can be collected at all seasons of the year, which represents a benefit for the industry.

Grapevine woody tissues accumulate stilbenoids following bud burst.

MAIN CONCLUSION: After bud burst, a transcriptional reprogramming of the shikimate and phenylpropanoid pathways occurs in grapevine canes resulting in the accumulation of stilbenoids like resveratrol and viniferin. Stilbenoids are phenylpropanoid compounds with important biological properties and biotechnological applications that are synthesized in grapevine in response to different stresses. Although they are found in woody tissues, such as canes and buds, their biosynthesis and accumulation have been essentially described in berries. We have previously shown that transcripts encoding secondary metabolism enzymes accumulate in grapevine canes following the transition from dormancy (E-L 1) to bud burst (E-L 4) suggesting that secondary metabolites may accumulate in grapevine canes during this transition. In the present study, using UPLC-MS we demonstrate the accumulation of important metabolites such as ferulic acid and the stilbenoids E-resveratrol, E-piceatannol and E-ε-viniferin. Stilbenoids accumulation correlated with the increased expression of several stilbene synthase genes and of VviMYB14, encoding a transcription factor that regulates stilbene biosynthesis. In addition, a general stimulation of the plastidial shikimate pathway was observed. Taken together, results show that important secondary metabolites accumulate in the woody canes during bud burst. These findings may aid biotechnological approaches aimed at extracting biologically active phenolic compounds, including stilbenoids, from grapevine woody tissues.

Evaluation of biotechnological processing through solid-state fermentation of oilseed cakes on extracts bioactive potential.

Oilseed cakes (OC) are natural sources of lignocellulosic biomass, produced every year in large amounts. In addition to their main applications as animal feed, plant or soil fertilizer, and compost, they present enormous potential for being used in biotechnological processes for the obtainment and extraction of valuable bioactive compounds. This work evaluated the effect of solid-state fermentation on the bioactive properties of extracts obtained from the bioprocessing of OC and evaluated the effect of solvents on the recovery of compounds with higher bioactive potential. A general decrease of EC50 values was observed for fermented extracts obtained using a mixture of water/methanol (1:1) as extraction solvent. A decrease in the minimum inhibitory concentration was observed for fermented water extracts compared to non-fermented. Additionally, growth inhibition of Listeria monocytogenes was observed when using aqueous methanolic fermented extracts. These extracts also exhibited a higher percentage of growth reduction against phytopathogenic fungi, and some extracts exhibited increased protection against genotoxic agents such as camptothecin and bisphenol A. It was demonstrated that bioprocessing of OC through SSF is an effective approach to obtaining valuable compounds with bioactive properties for use in the food, pharmaceutical or cosmetic industries.

Opuntia ficus-indica Fruit: A Systematic Review of Its Phytochemicals and Pharmacological Activities.

The use of Opuntia ficus-indica fruits in the agro-food sector is increasing for a multiplicity of players. This renewed interest is, in part, due to its organoleptic characteristics, nutritional value and health benefits. Furthermore, industries from different sectors intend to make use of its vast array of metabolites to be used in different fields. This trend represents an economic growth opportunity for several partners who could find new opportunities exploring non-conventional fruits, and such is the case for Opuntia ficus-indica. O. ficus-indica originates from Mexico, belongs to the Cactaceae family and is commonly known as opuntia, prickly pear or cactus pear. The species produces flowers, cladodes and fruits that are consumed either in raw or in processed products. Recent publications described that consumption of the fruit improves human health, exhibiting antioxidant activity and other relevant pharmacological activities through enzymatic and non-enzymatic mechanisms. Thus, we provide a systematic, scientific and rational review for researchers, consumers and other relevant stakeholders regarding the chemical composition and biological activities of O. ficus-indica fruits.

Bioprocessing of oilseed cakes by fungi consortia: Impact of enzymes produced on antioxidants release.

Oilseed cakes (OC) present high potential as feedstock for the biobased industry. Biotechnological processes allow OC valorization by the production of diverse value-added products and simultaneously altering OC structure, improving their nutritional value, and boosting OC utilization in animal feed. This work explored the use of fungi consortium of two different species as a bioprocessing approach to improve the nutritional quality of OC, obtain enzymes and antioxidants by solid-state fermentation (SSF) of sunflower cake (SFC) and rapeseed cake (RSC). Rhyzopus oryzae and Aspergillus ibericus consortium led to the highest production of cellulase (135 U/g) and ß-glucosidase (265 U/g) while maximum protease (228 U/g) was obtained with A. niger and R. oryzae consortium. Maximum xylanase production (886 U/g) was observed in SSF of RSC resulting in high hemicellulose reduction. The synergistic action of lignocellulosic enzymes resulted in extracts with increased antioxidant potential with possible application as food additives against oxidative stress.

Monoclonal antibody based immunoassay: An alternative way for aquatic environmental selenium detection.

Environmental concerns about human health encouraged increasing methodological interest in selenium (Se), which is an essential non-metal trace element and varies within a narrow concentration range between essential and toxic. In this study, two types of long-armed Se haptens (Se-hapten-lc-NHS) were synthesized for the first time using active ester formalization. In producing monoclonal antibodies (mAbs), the derivatization of haptenized Se at para- (meta-) and ortho-sites showed different properties. Finally, a mAb derived from hybridoma 5A52 was confirmed to be capable of establishing an indirect competitive enzyme-linked immunosorbent assay (ic-ELISA). There was a successful quantitative determination of Se4+ with a detection range of 17 to 207 pmol mL-1 and a limit of detection of approximately 3.9 pmol mL-1. The mAb was found to be remarkably sensitive and specific, with no evidence of cross-reactivity with other ions. The assay was validated for four kinds of Se forms in water samples and showed satisfactory recoveries between 80 % and 108 %, with coefficients of variation of 2.1 %-11 %. The method proposed in our study offers a useful protocol for the rapid screening of Se and provides an alternative solution for the analysis of Se in aquatic environments.

Hypericum perforatum and Its Potential Antiplatelet Effect.

BACKGROUND: Hypericum perforatum (HP) is currently one of the most consumed medicinal plants in the world. In traditional Chinese medicine, the herb hypericum (Guan Ye Lian Qiao) belongs to the group of plants that clarify heat. It is also used to treat various types of infection and inflammation. In contrast to the extensive literature on the antidepressant effects of HP, little is known about its action on platelets. The main objective of this work was to investigate the possible relevance of HP to platelet function. METHODS: We characterized the profile of platelet activation in the presence of HP extracts through an evaluation of molecular markers by flow cytometry: mobilization of intracellular Ca++ and expression of platelet receptors such as activated GPIIbIIIa and P-selectin (CD62). RESULTS: The results indicated a possible inhibitory effect of HP on the platelet activation response, which could be explained by the effect on intracellular calcium mobilization and the expression of activated GPIIbIIIa receptors. Despite of the limitations of an in vitro study, our results provide evidence of the possible mechanisms of action of HP. CONCLUSIONS: Further studies are needed to elucidate the effect of HP on hemostasis, but it may be recognized as a substance with antiplatelet properties.

Protection against Paraquat-Induced Oxidative Stress by Curcuma longa Extract-Loaded Polymeric Nanoparticles in Zebrafish Embryos.

The link between oxidative stress and environmental factors plays an important role in chronic degenerative diseases; therefore, exogenous antioxidants could be an effective alternative to combat disease progression and/or most significant symptoms. Curcuma longa L. (CL), commonly known as turmeric, is mostly composed of curcumin, a multivalent molecule described as having antioxidant, anti-inflammatory and neuroprotective properties. Poor chemical stability and low oral bioavailability and, consequently, poor absorption, rapid metabolism, and limited tissue distribution are major restrictions to its applicability. The advent of nanotechnology, by combining nanosacale with multi-functionality and bioavailability improvement, offers an opportunity to overcome these limitations. Therefore, in this work, poly-Ɛ-caprolactone (PCL) nanoparticles were developed to incorporate the methanolic extract of CL, and their bioactivity was assessed in comparison to free or encapsulated curcumin. Their toxicity was evaluated using zebrafish embryos by applying the Fish Embryo Acute Toxicity test, following recommended OECD guidelines. The protective effect against paraquat-induced oxidative damage of CL extract, free or encapsulated in PCL nanoparticles, was evaluated. This herbicide is known to cause oxidative damage and greatly affect neuromotor functions. The overall results indicate that CL-loaded PCL nanoparticles have an interesting protective capacity against paraquat-induced damage, particularly in neuromuscular development that goes well beyond that of CL extract itself and other known antioxidants.

How Does Hedonic Aroma Impact Long-Term Anxiety, Depression, and Quality of Life in Women with Breast Cancer? A Cross-Lagged Panel Model Analysis.

Depression and anxiety are common symptoms during and after adjuvant chemotherapy treatment for breast cancer (BC), with implications on quality of life (QoL). The present study evaluates the temporal relationship between anxiety, depression, and QoL (primary outcomes), as well as the impact of hedonic aroma (essential oils) on this relationship. This is a secondary analysis of a previously reported randomized controlled trial, with two groups: an experimental group (n = 56), who were subjected to the inhalation of a self-selected essential oil during chemotherapy, and a control group (n = 56), who were only subjected to the standard treatment. The hedonic aroma intervention occurred in the second (T1), third (T2), and fourth (T3) chemotherapy sessions, three weeks apart from each other. The follow-up (T4) assessments took place three months after the end of the treatment. Cross-lagged panel models were estimated in the path analysis framework, using structural equation modeling methodology. Regarding the control group, the cross-lagged panel model showed that anxiety at T1 predicted anxiety at T3, which in turn predicted both QoL and depression at T4. In the experimental group, hedonic aroma intervention was associated with stability of anxiety and QoL over time from T1 to T3, with no longitudinal prediction at T4. For women undergoing standard chemotherapy treatment, anxiety was the main longitudinal precursor to depression and QoL three months after chemotherapy. Thus, essential oils could complement chemotherapy treatment for early-stage BC as a way to improve long-term emotional and QoL-related adjustment.

Development of quantum dot-linked immunosorbent assay (QLISA) and ELISA for the detection of sunset yellow in foods and beverages.

Sunset yellow (SY) is widely used as food colorant. Excess and illegal use of SY could pose potential health risk. Quantum dots have been successfully used in biological research due to the high photoluminescence and high resistance to photobleaching. To analyze SY efficiently, quantum dot-linked immunosorbent assay (QLISA) and enzyme-linked immunosorbent assay (ic-ELISA) were developed on the basis of generated monoclonal antibody. A carboxyl group was introduced to SY and coupled with carrier proteins to synthesize artificial antigen. Under the optimal conditions, inhibitory concentrations (IC50) of SY were 1.9 ng/mL (ic-ELISA) and 3.4 ng/mL (QLISA); the limits of detection (LODs) were 0.2 ng/mL (ic-ELISA) and 1.0 ng/mL (QLISA), respectively. Cross-reactivities of the mAb toward eight kinds of analogues were<0.01%. The recovery rates in spiked foods and beverages were 75.6%∼120.1% (ic-ELISA) and 74.0%∼114.1% (QLISA), respectively.

Quercetin-biapigenin nanoparticles are effective to penetrate the blood-brain barrier.

Search for efficient therapeutic agents for central nervous system (CNS) disorders has been extensive. Nevertheless, blood-brain barrier (BBB) is an obstacle that prevents the majority of compounds to act in these diseases. It is, thus, of extreme relevance the BBB overcome, in order to deliver a drugs therapeutically active concentration to the action site, with the least losses and interaction with other organs, tissues, or cells. The present study aimed to investigate the potential protective effect of quercetin-biapigenin encapsulated into poly(Ɛ-polycaprolactone) (PCL) nanoparticles against t-BOOH-induced oxidative stress in several brain cell lines, as well as evaluate the permeability of those active molecules through an in vitro BBB model. The three cell lines under study (BV-2, hcmec/D3, and U87) presented different reactions to t-BOOH. In general, quercetin-biapigenin PCL-loaded nanoparticles were able to minimize compound toxicity they convey, regardless the cell line. Quercetin-biapigenin PCL-loaded nanoparticles (Papp of approximately 80 × 10-6 cm/s) revealed to be more permeable than free compounds (Papp of approximately 50 × 10-6 cm/s). As of our knowledge, this is the first report of quercetin-biapigenin PCL-loaded nanoparticle activity in brain cells. It is also the first determining its permeability through BBB, as an effective nanocarrier for brain delivery.

Degradation of lignocellulosic matrix of oilseed cakes by solid-state fermentation: fungi screening for enzymes production and antioxidants release.

BACKGROUND: Vegetable oils are yearly produced in large amounts generating solid by-products, the oilseed cake (OC). OCs are lignocellulosic materials that have been used for animal feed with some limitations due to high fibre content from the plant cell walls. Biotechnological processes can help to overcome these limitations and contribute to up-grading such by-products, enhancing their nutritional value as feed ingredients. RESULTS: All fungal species were able to decrease neutral detergent fibre and acid detergent fibre in all by-products. Additionally, relevant enzymes were produced by the three fungi studied resulting in an improved antioxidant capacity of all fermented OCs. Aspergillus niger led to the highest activity of cellulase (109 U g-1 ), xylanase (692 U g-1 ) and protease (157 U g-1 ) per dry OC matter and to the recovery of an extract rich in antioxidants, with the highest scavenging potential of free radicals and superoxide anion, iron chelation ability and reducing power. Rhyzopus oryzae produced the highest activity of ß-glucosidase (503 U g-1 ) and led to the highest liberation of total phenolic content (TPC). Principal components analysis showed that extracts with high antioxidant potential were obtained in solid-state fermentation (SSF) with high enzymatic activity. A positive correlation was established between the action of ß-glucosidase and TPC. CONCLUSION: Within the same bioprocess it was possible to improve the nutritional value of OCs and to obtain relevant bioactive compounds such as lignocellulosic enzymes and phenolic compounds with antioxidant potential, resulting in a significant improvement of already valuable by-products with commercial interest for animal feed. © 2021 Society of Chemical Industry.

HPLC-DAD Fingerprints Combined With Multivariate Analysis of Epimedii Folium From Major Producing Areas in Eastern Asia: Effect of Geographical Origin and Species.

The growth location and plant variety may influence the active components and biological activities of plants used in phytomedicine. In this study, nine sets of different Epimedii Folium, from different representative cultivation locations and Epimedium species, were collected for comparison, using HPLC-DAD combined with multivariate analysis. The objective was to investigate the influence of geographical origin and Epimedium species on the quality of Epimedii Folium, and provide applicable guidance for cultivation and quality control of Epimedii Folium. Several Epimedium spp. sets were used to establish the HPLC-DAD fingerprints and 91 peaks (compounds) were selected for the multivariate analysis. Major compounds were analyzed by HPLC-DAD combined with principal component analysis (PCA). HPLC quantitative analysis of known bioactive compounds was performed. Application of PCA to HPLC data showed that Epimedium samples sharing the same geographical origin or species clustered together, indicating that both species and geographical origin have impacts on the quality of Epimedii Folium. The major bioactive flavonoid compounds, epimedin C, icariin and baohuoside I, were identified and quantified. The concentration of bioactive compounds was significantly influenced both by species and geographical origin. E. sagittatum from Sichuan showed the highest content of bioactive compounds. The results showed that both Epimedium species and geographical origin have strong impact into quality of Epimedii Folium. HPLC data combined with multivariate analysis is a suitable approach to inform the selection of cultivation areas and choose Epimedium spp. most suitable for different geographical areas, resulting in improved quality of Epimedii Folium.

In vivo anti-inflammatory and antinociceptive effects, and in vitro antioxidant, antiglycant and anti-neuroinflammatory actions of Syzygium malaccense.

Syzygium malaccense is popularly used to treat inflammation and pain-related ailments. The species was assessed regarding its antioxidant, antiglycant, anti-inflammatory, including anti-neuroinflammatory, and antinociceptive activities. Different models were employed to measure S. malaccense extract (ESM) antioxidant activity. The antiglycant activity was determined using the glucose-induced protein glycation model. LPS-induced neuroinflammation on murine BV-2 microglial cell line was used for anti-neuroinflammatory activity evaluation. The croton oil-induced ear edema test was accomplished to evaluate the in vivo anti-inflammatory activity. Acetic acid-induced writhing together with formalin-induced paw licking assays were performed to evaluate the antinociceptive potential. Finally, the chemical characterization was accomplished by a UHPLC-MS analysis. ESM presented relevant antioxidant and antiglycant activity. NO production by BV-2 cells was reduced, indicating the relevant neuroprotective activity. ESM significantly decreased the mice ear edema induced by croton oil and the nociceptive stimulus induced by acetic acid and formalin by central and peripheral mechanisms. The flavonoids myricitrin, myricetin and quercetin were identified and, as far as we know, the alkaloid reserpine was reported in the species for the first time. The antioxidant and antiglycant potential of ESM, may be related to the in vivo anti-inflammatory and antinociceptive effects, and to the in vitro neuroinflammation inhibition.

Transcriptional responses of Hypericum perforatum cells to Agrobacterium tumefaciens and differential gene expression in dark glands.

Hypericum perforatum L. (St. John's wort) is a well-known medicinal plant that possesses secondary metabolites with beneficial pharmacological properties. However, improvement in the production of secondary metabolites via genetic manipulation is a challenging task as H. perforatum remains recalcitrant to Agrobacterium tumefaciens-mediated transformation. Here, the transcripts of key genes involved in several plant defence responses (secondary metabolites, RNA silencing, reactive oxygen species (ROS) and specific defence genes) were investigated in H. perforatum suspension cells inoculated with A. tumefaciens by quantitative real-time PCR. Results indicated that key genes from the xanthone, hypericin and melatonin biosynthesis pathways, the ROS-detoxification enzyme HpAOX, as well as the defence genes Hyp-1 and HpPGIP, were all upregulated to rapidly respond to A. tumefaciens elicitation in H. perforatum. By contrast, expression levels of genes involved in hyperforin and flavonoid biosynthesis pathways were markedly downregulated upon A. tumefaciens elicitation. In addition, we compared the expression patterns of key genes in H. perforatum leaf tissues with and without dark glands, a major site of secondary metabolite production. Overall, we provide evidence for the upregulation of several phenylpropanoid pathway genes in response to elicitation by Agrobacterium, suggesting that production of secondary metabolites could modulate H. perforatum recalcitrance to A. tumefaciens-mediated transformation.