[Analysis of constituents in different parts of Forsythia suspensa by UPLC-Q-TOF-MS and evaluation of their anti-inflammatory activity].

This study aims to characterize and identify the chemical constituents in 11 parts of Forsythia suspensa by using ultra-performance liquid chromatography-quadrupole time of flight-mass spectrometry(UPLC-Q-TOF-MS) combined with a self-established chemical constituent database, including leaves, flowers, fruits, green F. suspensa, old F. suspensa, and seeds. The quality attributes and differences of different parts of F. suspensa were evaluated by principal component analysis, partial least square discriminant analysis, and other stoichiometric methods. A total of 79 compounds were identified, including 13 phenylethanol glycosides, 10 lignans, 12 flavonoids, 10 organic acids, 14 terpenoids, and 20 other types of compounds. Among them, 34 compounds were the main variables of difference between the different parts of F. suspensa, and the content of each component was relatively higher in the leaves and green F. suspensa. The LPS-induced inflammation model of RAW264.7 cells was applied to study the anti-inflammatory activity of the extracts of the different parts of F. suspensa and the main constituents. The results show that the extracts of green F. suspensa, flower, twig, and stem exhibited anti-inflammatory activity, and the constituents such as forsythoside A, phyllyrin, phillygenin, and(+)-pinoresinol-ß-D-glucopyranoside could significantly inhibit anti-inflammatory activity released by NO. The chemical constituent in different parts of F. suspensa is analyzed comprehensively, and the anti-inflammatory activity is evaluated in this study, which provides a reference for the development and comprehensive utilization of F. suspensa resources.

The phytochemistry, pharmacology, pharmacokinetics, quality control, and toxicity of Forsythiae Fructus: An updated systematic review.

Forsythiae Fructus (FF), the dried fruit of F. suspensa, is commonly used to treat fever, inflammation, etc in China or other Asian countries. FF is usually used as the core herb in traditional Chinese medicine preparations for the treatment of influenza, such as Shuang-huang-lian oral liquid and Yin-qiao powder, etc. Since the wide application and core role of FF, its research progress was summarized in terms of traditional uses, phytochemistry, pharmacology, pharmacokinetics, quality control, and toxicity. Meanwhile, the anti-influenza substances and mechanism of FF were emphasized. Till now, a total of 290 chemical components are identified in F. suspensa, and among them, 248 components were isolated and identified from FF, including 42 phenylethanoid glycosides, 48 lignans, 59 terpenoids, 14 flavonoids, 3 steroids, 24 cyclohexyl ethanol derivatives, 14 alkaloids, 26 organic acids, and 18 other types. FF and their pure compounds have the pharmacological activities of anti-virus, anti-inflammation, anti-oxidant, anti-bacteria, anti-tumor, neuroprotection, hepatoprotection, etc. Inhibition of TLR7, RIG-I, MAVS, NF-κB, MyD88 signaling pathway were the reported anti-influenza mechanisms of FF and phenylethanoid glycosides and lignans are the main active groups. However, the bioavailability of phenylethanoid glycosides and lignans of FF in vivo was low, which needed to be improved. Simultaneously, the un-elucidated compounds and anti-influenza substances of FF strongly needed to be explored. The current quality control of FF was only about forsythoside A and phillyrin, more active components should be taken into consideration. Moreover, there are no reports of toxicity of FF yet, but the toxicity of FF should be not neglected in clinical applications.

Assembly and analysis of the complete mitochondrial genome of Forsythia suspensa (Thunb.) Vahl.

BACKGROUND: Forsythia suspensa (Thunb.) Vahl is a valuable ornamental and medicinal plant. Although the nuclear and chloroplast genomes of F. suspensa have been published, its complete mitochondrial genome sequence has yet to be reported. In this study, the genomic DNA of F. suspensa yellowish leaf material was extracted, sequenced by using a mixture of Illumina Novaseq6000 short reads and Oxford Nanopore PromethION long reads, and the sequencing data were assembled and annotated. RESULT: The F. suspensa mitochondrial genome was obtained in the length of 535,692 bp with a circular structure, and the GC content was 44.90%. The genome contains 60 genes, including 36 protein-coding genes, 21 tRNA genes, and three rRNA genes. We further analyzed RNA editing of the protein-coding genes, relative synonymous codon usage, and sequence repeats based on the genomic data. There were 25 homologous sequences between F. suspensa mitochondria and chloroplast genome, which involved the transfer of 8 mitochondrial genes, and 9473 homologous sequences between mitochondrial and nuclear genomes. Analysis of the nucleic acid substitution rate, nucleic acid diversity, and collinearity of protein-coding genes of the F. suspensa mitochondrial genome revealed that the majority of genes may have undergone purifying selection, exhibiting a slower rate of evolution and a relatively conserved structure. Analysis of the phylogenetic relationships among different species revealed that F. suspensa was most closely related to Olea europaea subsp. Europaea. CONCLUSION: In this study, we sequenced, assembled, and annotated a high-quality F. suspensa mitochondrial genome. The results of this study will enrich the mitochondrial genome data of Forsythia, lay a foundation for the phylogenetic development of Forsythia, and promote the evolutionary analysis of Oleaceae species.

New lignans and phenylethanoid with antioxidant activity from aerial parts of Forsythia suspensa (Thunb.) Vahl.

Two new lignans, phillyroside A(1) and phillyroside B(2), together with three new phenylethanoid glycoside, forsythoside K(3), forsythoside L(5) and forsythol L (4), while compounds 4 was an aglycon of forsythoside L(5), were isolated from the aerial parts of Forsythia suspensa (Thunb.) Vahl. Their structures were elucidated by comprehensive analyses of standard spectroscopic data (MS, IR, and NMR) and the in vitro antioxidant activity of five new compounds were evaluated in DPPH and ABTS radical scavenging experiment and ferric reducing ability of plasma (FRAP) experiment. Compounds 4 and 5 exhibited antioxidant activity with IC50 values ranging from 112.49 to 153.58 µM in DPPH experiment and 45.43 to 64.09 µM in ABTS experiment.

Green Extraction of Forsythoside A, Phillyrin and Phillygenol from Forsythia suspensa Leaves Using a ß-Cyclodextrin-Assisted Method.

In this study, a green process of ß-cyclodextrin (ß-CD)-assisted extraction of active ingredients from Forsythia suspensa leaves was developed. Firstly, the optimal process of extraction was as follows: the ratio between Forsythia suspensa leaves and ß-CD was 3.61:5, the solid-liquid ratio was 1:36.3, the temperature was 75.25 °C and the pH was 3.94. The yields of forsythoside A, phillyrin and phillygenol were 11.80 ± 0.141%, 5.49 ± 0.078% and 0.319 ± 0.004%, respectively. Then, the structure characteristics of the ß-CD-assisted extract of Forsythia suspensa leaves (FSE-ß-CD) were analyzed using powder X-ray diffraction (PXRD), Fourier transform infrared spectroscopy (FT-IR), differential scanning calorimetry (DSC), scanning electron microscopy (SEM) and molecular docking to demonstrate that the natural active products from Forsythia suspensa leaves had significant interactions with the ß-CD. Additionally, the loss of forsythoside A from aqueous FSE-CD at 80 °C was only 12%, compared with Forsythia suspensa leaf extract (FSE) which decreased by 13%. In addition, the aqueous solubility of FSE-CD was significantly increased to 70.2 g/L. The EC50 for scavenging DPPH and ABTS radicals decreased to 28.98 ug/mL and 25.54 ug/mL, respectively. The results showed that the ß-CD-assisted extraction process would be a promising technology for bioactive compounds extracted from plants.

Visualizing the spatial distribution of functional metabolites in Forsythia suspensa at different harvest stages by MALDI mass spectrometry imaging.

As a traditional Chinese medicine, Forsythia suspensa (F. suspensa) has attracted much attention due to its significant pharmacological activity. Revealing the spatial distribution of metabolites during F. suspensa development is important for understanding its biosynthesis rules and improving the quality of medicinal materials. However, there is currently a lack of information on the spatial distribution of F. suspensa metabolites. In this work, the spatial distribution and growth metabolism patterns of important metabolites of F. suspensa were studied for the first time using matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI). Using 2,5-dimethylnaphthalene (DAN) as the matrix and detecting in negative ion mode, the spatial distribution and growth patterns of 11 metabolites obtained from longitudinal sections of F. suspensa included pinoresinol, phillygenin, forsythoside A, forsythoside E, rutin, caffeic acid, malic acid, citric acid, stearic acid, oleic acid, and linoleic acid. These results showed the mesocarp and endosperm tissues of F. suspensa were important for storing important functional metabolites. Changes in mesocarp and endosperm growth and development tissues caused large changes in the content of important functional metabolites in F. suspensa. These results provide a basis for understanding the spatial distribution of metabolites in F. suspensa tissues and the significant changes that occur during growth and development, exploring the mechanism of important synthesis of metabolites, regulating the harvest of F. suspensa, and improving the quality of medicinal herbs.

Metabolome analysis of genus Forsythia related constituents in Forsythia suspensa leaves and fruits using UPLC-ESI-QQQ-MS/MS technique.

Forsythia suspensa is a traditional Chinese herb. Its numerous metabolites have important roles, as they possessed a wide range of biological activities. This study explored the accumulations of F. suspensa metabolites by performing widely targeted metabolomic analysis. The metabolites were studied at four stages of fruit development. Metabolites in the fruits and leaves of F. suspensa during fruit development included phenolic acids, flavonoids, lipids, lignans and coumarins, amino acids and their derivatives, terpenes, organic acids, nucleotides and their derivatives, alkaloids, quinones, steroids, and tannins. Fourteen Forsythia related metabolites were detected. Their contents varied among the developmental stages. Statistically significant correlations were found between the levels of forsythoside B and 11-methyl-forsythide, and forsythialan B and phillygenin, in both leaves and fruits. According to the correlation analysis between metabolites, Forsythia related metabolites were divided into two classes and five subclasses. In total, 33 compounds presented significant correlations in both fruits and leaves, which indicated the potential relationship in the synthesis of Forsythia related metabolites. Forsythialan B and phillygenin were both negatively correlated with L-valine, while Z-6,7-epoxyligustilid was positively correlated with both compounds. The quality control compounds forsythiaside A and phillyrin were positively and negatively correlated with uracil, respectively. These metabolomics results may facilitate the biosynthesis of Forsythia related metabolites.

Identification of Forsythia suspensa (Thunb.) Vahl in different harvest periods using intelligent sensory technologies, HPLC characteristic fingerprint coupled with chemometrics.

INTRODUCTION: Forsythia suspensa (Thunb.) Vahl (FS), the fruit of Oleaceae plants, as a large part of traditional Chinese medicine, is classified as "Qingqiao (Q)" and "Laoqiao (L)" based on the harvest time. Because the maturation of FS is a gradual process, its accurate identification based on different maturity levels is an important issue. OBJECTIVES: We suggest colorimetric, electronic tongue, and high-performance liquid chromatography (HPLC) characteristic fingerprints to discriminate FS in different harvest periods. MATERIAL AND METHODS: First, FS fruits from different harvest times were collected, and then, their colour parameters, E-tongue sensory properties, HPLC characteristic fingerprints, and contents of nominal ingredients were determined. Finally, multivariate statistical analyses, including three-dimensional scatter plots, hierarchical cluster, principal component, linear discriminant, similarity, and partial least squares discriminant analyses were performed. RESULTS: The results demonstrated that the three experimental techniques could effectively discriminate FS based on different harvest times with 100% accuracy. Under the qualitative conditions, nine common peaks were identified in the HPLC fingerprints of 60 samples, among which, six peaks [variable importance in projection (VIP) > 1] could be used as index peaks for qualitative identification. In fact, the contents of quality marker components, including forsythin, phillygenin, rutin and forsythoside A, were significant different (P < 0.001) at different harvest times. Interestingly, the quality markers not only accurately reflected the maturity of FS but also showed close correlations with the colour parameters and sensory E-tongue responses. CONCLUSION: In our present investigation, bionic technologies, including a colorimeter, E-tongue analysis, and HPLC characteristic fingerprints, combined with chemometrics, were employed to develop a novel and accurate method for discriminating FS based on different harvest times.

Protective effects of Forsythiae fructus and Cassiae semen water extract against memory deficits through the gut-microbiome-brain axis in an Alzheimer's disease model.

CONTEXT: Fruits of Forsythia suspensa Vahl (Oleaceae) and seeds of Cassia obtusifolia Linne (Caesalpinaceae) have been used to treat inflammation in Asia. OBJECTIVE: We examined the alleviation of memory function in Alzheimer's disease (AD) rats fed Forsythiae Fructus (FF) and Cassiae Semen water extracts (CS) and investigated the mechanisms responsible for the effects. MATERIALS AND METHODS: Thirty Sprague-Dawley male rats had hippocampal infusions of amyloid-ß(25-35) (AD rats; memory deficit), and ten rats were infused with amyloid-ß(35-25) (non-AD rats; no memory deficit). For eight weeks, all rats freely consumed high-fat diets (43% lard) incorporated with 200 mg/kg body weight assigned aqueous herbal extracts: AD-FF, AD-CS, or without extracts AD-CON (control), non-AD (normal-control). RESULTS: Memory impairment was prevented in the AD-FF (0.54 ± 0.06-fold) and the AD-CS rats (0.33 ± 0.04-fold) compared to the AD-CON by inhibiting amyloid-ß deposition to the levels less than one-fourth of the AD-CON group. The hippocampal pAkt→pGSK-3ß→pFOXO1 pathway was attenuated by approximately 3.25-fold in the AD-CON, while AD-FF prevented the attenuation better than AD-CS. The relative intensity of hippocampal tau protein based on ß-actin was suppressed with AD-FF (0.68 ± 0.09) and AD-CS (0.96 ± 0.81), compared to AD-CON (1.19 ± 0.13). AD decreased the abundance of Bacteroidales by 34.2% and Lactobacillales by 23.8% and increased Clostridiales by 181% while the AD-FF, but not the AD-CS, normalised the gut microbiota changes to be similar to the non-AD. DISCUSSION AND CONCLUSIONS: FF improved memory deficits better than CS in an AD-induced rat model. The potential neuroprotective benefits of FF against AD may be applicable to human AD therapy with additional clinical research.

A new diterpernoid glycoside from the fruit of Forsythia suspensa.

One new diterpernoid glycoside, lanceolatinoside A (1), together with one known iridoid glycoside, luzonoside C (2) were isolated from the fruit of Forsythia suspensa. The structure of the new compound (1) was elucidated through 1 D and 2 D NMR spectroscopic data, and HR-ESIMS. Compound 2 was identified as luzonoside C (2) on the basis of NMR spectroscopic data analyses and comparison with those reported in the literature.

New C9 -Monoterpenoid Alkaloids Featuring a Rare Skeleton with Anti-Inflammatory and Antiviral Activities from Forsythia suspensa.

Forsyqinlingines C (1) and D (2), two C9 -monoterpenoid alkaloids bearing a rare skeleton, were isolated from the ripe fruits of Forsythia suspensa. Their structures, including absolute configurations, were fully elucidated by extensive spectroscopic data and ECD experiments. The plausible biogenetic pathway for compounds 1 and 2 was also proposed. In vitro, two C9 -monoterpenoid alkaloids showed anti-inflammatory activity performed by the inhibitory effect on the release of ß-glucuronidase in rat polymorphonuclear leukocytes (PMNs), as well as antiviral activity against influenza A (H1N1) virus and respiratory syncytial virus (RSV).

Transmembrane G protein-coupled receptor 5 signaling stimulates fibroblast growth factor 21 expression concomitant with up-regulation of the transcription factor nuclear receptor Nr4a1.

Fibroblast growth factor 21 (FGF21) acts as an endocrine factor, playing important roles in the regulation of energy homeostasis, glucose and lipid metabolism. It is induced by diverse metabolic and cellular stresses, such as starvation and cold challenge, which in turn facilitate adaptation to the stress environment. The pharmacological action of FGF21 has received much attention, because the administration of FGF21 or its analogs has been shown to have an anti-obesity effect in rodent models. In the present study, we found that 3-O-acetyloleanolic acid, an active constituent isolated from the fruits of Forsythia suspensa, stimulated FGF21 production concomitant with the up-regulation of a transcription factor, nuclear receptor Nr4a1, in C2C12 myotubes. Additionally, significant increases in mFgf21 promoter activity were observed in C2C12 cells overexpressing TGR5 receptor in response to 3-O-acetyloleanolic acid treatment. Treatment with the p38 MAPK inhibitor SB203580 was effective at suppressing these stimulatory effects of 3-O-acetyloleanolic acid. Pretreatment with SB203580 also significantly repressed FGF21 mRNA abundance and FGF21 secretion in C2C12 myotubes after 3-O-acetyloleanolic acid stimulation, suggesting that p38 activation is required for the induction of FGF21 by ligand-activated TGR5 in C2C12 myotubes. These findings collectively indicated that TGR5 receptor signaling drives FGF21 expression via p38 activation, at least partly, by mediating Nr4a1 expression. Thus, the novel biological function of 3-O-acetyloleanolic acid as an agent having anti-obesity effects is likely to be mediated through the activation of TGR5 receptors.

Forsythia Fruit Prevents Fulminant Hepatitis in Mice and Ameliorates Inflammation in Murine Macrophages.

Forsythia Fruit (FF), the fruit of Forsythia suspensa, has been used since ancient times as an herbal medication in East Asia to treat inflammation, gonorrhea, and pharyngitis. However, the efficacy of FF against liver damage due to inflammation has not been studied. Here, we explored the protective effects of FF in a mouse hepatitis model induced by lipopolysaccharide (LPS)/D-galactosamine (GalN) treatment. We measured inflammatory cytokine and aminotransferase levels in mouse blood and analyzed the effects of FF on inflammatory gene and protein expression levels in liver tissue. Our results show that FF treatment effectively lowers inflammatory cytokine and serum aminotransferase levels in mice and inhibits the expression of hepatic cytokine mRNA and inflammatory proteins. Furthermore, treatment with FF activated the antioxidant pathway HO-1/Nrf-2 and suppressed severe histological alteration in the livers of LPS/D-GalN-treated mice. Further investigation of the effects of FF on inflammatory reactions in LPS-stimulated macrophages showed that pretreatment with FF inhibits inflammatory mediator secretion and activation of inflammatory mechanisms both in a mouse macrophage RAW 264.7 cells and in primary peritoneal macrophages. These results show that FF has potential worth as a candidate for the treatment of fulminant inflammatory reactions and subsequent liver injury.

Forsythiae Fructus aqueous extract attenuates cisplatin-induced kaolin consumption (pica) by inhibiting NLRP3 inflammasome activation in rats.

The present study was conducted to evaluate the effect of Forsythiae Fructus aqueous extract (FAE) against cisplatin-induced emesis and to explore the antiemetic mechanism of FAE by focusing on NLRP3 inflammasome activation in a rat pica model. Our results showed that FAE significantly ameliorated cisplatin-induced acute and delayed pica in rats. Moreover, FAE improved the gastrointestinal histopathological injury and reduced the levels of serum ROS, IL-1ß, and IL-18 in cisplatin-treated rats. In addition, the expressions of NLRP3, ASC, caspase-1, and IL-1ß and the colocalization of the NLRP3 with ASC or caspase-1 in rat gastric antrum and ileum were also suppressed by FAE. Taken together, our findings indicate that FAE has a therapeutic effect against CINV, which may be related to its inhibition of the activation of NLRP3 inflammasome.

Neuroprotective and anti-inflammatory phenylethanoidglycosides from the fruits of Forsythia suspensa.

Neuroinflammation is emerging as a crucial reason of major neurodegenerative diseases in recent years. Increasingly evidences have supported that bioactive natural products from traditional Chinese medicines have efficiency for neuroinflammation. Forsythia suspensa, a typical medicinal herb, showed potential neuroprotective and anti-inflammatory properties in previous pharmacological studies. In our research to obtain neuroprotective and anti-inflammatory natural products, three unprecedented C6-C7'/C6-C16' linked phenylethanoidglycoside dimers (1-3), three new phenylethanoidglycosides (4-6), and six known compounds (7-12) were isolated from the fruits of Forsythia suspensa. Their structures were determined by comprehensive spectroscopic data and comparison to the literature data. All isolated compounds were evaluated their neuroprotective and anti-inflammatory activities. Compounds 1 and 10 exhibited significant neuroprotective activities with the cell viability values of 75.24 ± 8.05% and 93.65 ± 10.17%, respectively, for the serum-deprivation and rotenone induced pheochromocytoma (PC12) cell injury. Meanwhile, compound 1 exhibited excellent anti-inflammatory activity against tumor necrosis factor (TNF)-α expression in LPS induced RAW264.7 cells with the IC50 value of 1.30 µM. This study revealed that the bioactive phenylethanoidglycosides may attenuate neuroinflammation through their neuroprotective and anti-inflammatory activities.

A review of pharmacological and pharmacokinetic properties of Forsythiaside A.

Traditional Chinese medicine plays a significant role in the treatment of various diseases and has attracted increasing attention for clinical applications. Forsythiae Fructus, the dried fruit of Forsythia suspensa (Thunb.) Vahl, is a widely used Chinese medicinal herb in clinic for its extensive pharmacological activities. Forsythiaside A is the main active index component isolated from Forsythiae Fructus and possesses prominent bioactivities. Modern pharmacological studies have confirmed that Forsythiaside A exhibits significant activities in treating various diseases, including inflammation, virus infection, neurodegeneration, oxidative stress, liver injury, and bacterial infection. In this review, the pharmacological activities of Forsythiaside A have been comprehensively reviewed and summarized. According to the data, Forsythiaside A shows remarkable anti-inflammation, antivirus, neuroprotection, antioxidant, hepatoprotection, and antibacterial activities through regulating multiple signaling transduction pathways such as NF-κB, MAPK, JAK/STAT, Nrf2, RLRs, TRAF, TLR7, and ER stress. In addition, the toxicity and pharmacokinetic properties of Forsythiaside A are also discussed in this review, thus providing a solid foundation and evidence for further studies to explore novel effective drugs from Chinese medicine monomers.

Tissue-Specific Accumulation and Isomerization of Valuable Phenylethanoid Glycosides from Plantago and Forsythia Plants.

A comparative phytochemical study on the phenylethanoid glycoside (PhEG) composition of the underground organs of three Plantago species (P. lanceolata, P. major, and P. media) and that of the fruit wall and seed parts of Forsythia suspensa and F. europaea fruits was performed. The leaves of these Forsythia species and six cultivars of the hybrid F. × intermedia were also analyzed, demonstrating the tissue-specific accumulation and decomposition of PhEGs. Our analyses confirmed the significance of selected tissues as new and abundant sources of these valuable natural compounds. The optimized heat treatment of tissues containing high amounts of the PhEG plantamajoside (PM) or forsythoside A (FA), which was performed in distilled water, resulted in their characteristic isomerizations. In addition to PM and FA, high amounts of the isomerization products could also be isolated after heat treatment. The isomerization mechanisms were elucidated by molecular modeling, and the structures of PhEGs were identified by nuclear magnetic resonance spectroscopy (NMR) and high-resolution mass spectrometry (HR-MS) techniques, also confirming the possibility of discriminating regioisomeric PhEGs by tandem MS. The PhEGs showed no cytostatic activity in non-human primate Vero E6 cells, supporting their safe use as natural medicines and allowing their antiviral potency to be tested.

The Biological Effects of Forsythia Leaves Containing the Cyclic AMP Phosphodiesterase 4 Inhibitor Phillyrin.

Forsythia fruit (Forsythia suspensa Vahl (Oleaceae)) is a common component of Kampo medicines for treating the common cold, influenza, and allergies. The main polyphenolic compounds in the leaves of F. suspensa are pinoresinol ß-d-glucoside, phillyrin and forsythiaside, and their levels are higher in the leaves of the plant than in the fruit. It is known that polyphenolic compounds stimulate lipid catabolism in the liver and suppress dyslipidemia, thereby attenuating diet-induced obesity and polyphenolic anti-oxidants might attenuate obesity in animals consuming high-fat diets. Recently, phillyrin was reported as a novel cyclic AMP phosphodiesterase 4 (PDE4) inhibitor derived from forsythia fruit. It was expected that the leaves of F. suspensa might display anti-obesity effects and serve as a health food material. In this review, we summarized our studies on the biological effects of forsythia leaves containing phillyrin and other polyphenolic compounds, particularly against obesity, atopic dermatitis, and influenza A virus infection, and its potential as a phytoestrogen.

Antimicrobial effects of three herbs (Brassica juncea, Forsythia suspensa, and Inula britannica) on membrane permeability and apoptosis in Salmonella.

AIMS: This study aimed synergistic effects of three herbs in Salmonella via increased membrane permeability and apoptosis. METHODS AND RESULTS: Using high-performance liquid chromatography, four types of phenylethyl glycosides and a lignan were detected in the herb mixture (Brassica juncea, Forsythia suspensa, and Inula britannica). During treatment with the herb mixture (1×, 2×, or 4× the MIC), viable cells decreased to 1·87 log CFU per ml (Salmonella Gallinarum) and 2·33 log CFU per ml (Salmonella Enteritidis) after 12 h of incubation according to inhibition of tricarboxylic acid cycle (P < 0·01). In addition, N-phenyl-1-naphthylamine uptake increased from 229·00 to 249·67 AU in S. Gallinarum and from 232·00 to 250·67 AU in S. Enteritidis (P < 0·05), whereas membrane potential decreased from 8855·00 to 3763·25 AU and from 8703·67 to 4300·38 AU, respectively. Apoptotic Salmonella cells were observed by confocal laser scanning microscopy and flow cytometry. Transmission electron microscopy observations with negative staining showed protein leakage from damaged Salmonella. CONCLUSIONS: These results showed the synergistic effect of the three herbs against avian pathogenic Salmonella induced by membrane damage and apoptosis. SIGNIFICANCE AND IMPACT OF THE STUDY: Salmonella causes enormous economic losses in the poultry industry. These results indicated that potency of natural antimicrobial agents due to apoptosis in Salmonella.

Forsythia suspensa extract enhances performance via the improvement of nutrient digestibility, antioxidant status, anti-inflammatory function, and gut morphology in broilers.

This experiment aims to determine the effects of Forsythia suspense extract (FSE) as an antibiotic substitute on performance, antioxidant status, anti-inflammatory function, intestinal morphology, and meat fatty acid deposition in broilers. 192 male Arbor Acre broilers (1-day-old, weighing 45.6 ± 1.3 g) were randomly allocated to 3 treatments, 8 replicate pens per treatment, 8 broilers per pen. The treatments contain a control diet (corn-soybean meal basal diet, CTL), an antibiotic diet (basal diet + 75 mg/kg chlortetracycline, CTC), and an FSE diet (basal diet + 100 mg/kg FSE; FSE). The experiment includes phase 1 (day 1 to 21) and 2 (day 22 to 42). Compared with CTL and CTC, broilers supplemented with FSE showed higher (P < 0.05) ADG and ADFI in phase 2 and overall (day 1 to 42). On day 21, serum catalase and total antioxidant capacity contents were enhanced (P < 0.05) in broilers fed FSE compared with CTL. On day 42, broilers fed FSE showed increased (P < 0.05) serum superoxide dismutase and glutathione peroxidase contents, and enhanced (P < 0.05) apparent total tract digestibility of dry matter, organic matter, gross energy, total carbohydrates, and phosphorus, as well as reduced (P < 0.05) nitrogen and phosphorus excretion in feces compared with CTL. These broilers also showed decreased (P < 0.05) n-6/n-3 polyunsaturated fatty acid ratio in thigh meat, and tumor necrotic factor-alpha, interleukin-1ß and interleukin-6 contents in the liver on day 42 compared with CTL. The villus height was increased (P < 0.05) in the duodenum, jejunum, and ileum of broilers fed FSE compared with CTL. In conclusion, dietary F.suspense extract supplementation as a chlortetracycline substitute under non-challenge conditions enhanced performance via the improvement of nutrient digestibility, antioxidant status, anti-inflammatory function, and intestinal morphology in broilers. Moreover, F.suspense extract may also benefit environment by reducing nitrogen and phosphorus excretion and benefit human health via modulating meat fatty acid profiles in broilers.