Impact of ferulic acid and resveratrol on the effectiveness and safety of sunscreen

Abstract The combination of avobenzone (AVO) and octyl ρ-methoxycinnamate (OMC) is widely used to ensure broad-spectrum photo-protection because they absorb UVA and UVB, respectively. However, they are thermally and photo unstable because they degrade and undergo photo- tautomerization and trans-cis isomerization, thus reducing their photo-protection efficacy during UV exposure. This study aimed to evaluate the potential use of the antioxidants ferulic acid and resveratrol as stabilizing substances in AVO and OMC mixtures in solution or emulsion. The effects of both antioxidants on the thermal/photo-stability and suppression of the filter singlet state, besides skin permeation, were evaluated. Both antioxidants contributed to preserving OMC and AVO during the thermal stability test, which relates to the maintenance of photo-protection even after storing the formulations at high temperatures. Nevertheless, although resveratrol retained part of the OMC trans isomer and suppressed the AVO singlet state when exposed to UV, no contribution to photo-protection stability was observed, contrary to expectations. Regarding the permeation assay, the addition of both antioxidants was accompanied by a reduction of AVO permeation, while resveratrol increased OMC permeation. Thus, the chemical and physicochemical properties of these antioxidants impacted their efficacy and safety profiles; therefore, further studies are required to establish the real cost-benefit ratio for their use in sunscreens.

Ferulic acid enhances insulin secretion by potentiating L-type Ca2+ channel activation / 中西医结合学报

OBJECTIVE@#To investigate the effect of ferulic acid, a natural compound, on pancreatic beta cell viability, Ca2+ channels, and insulin secretion.@*METHODS@#We studied the effects of ferulic acid on rat insulinoma cell line viability using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide viability assay. The whole-cell patch-clamp technique and enzyme-linked immunosorbent assay were also used to examine the action of ferulic acid on Ca2+ channels and insulin secretion, respectively.@*RESULTS@#Ferulic acid did not affect cell viability during exposures up to 72 h. The electrophysiological study demonstrated that ferulic acid rapidly and concentration-dependently increased L-type Ca2+ channel current, shifting its activation curve in the hyperpolarizing direction with a decreased slope factor, while the voltage dependence of inactivation was not affected. On the other hand, ferulic acid have no effect on T-type Ca2+ channels. Furthermore, ferulic acid significantly increased insulin secretion, an effect inhibited by nifedipine and Ca2+-free extracellular fluid, confirming that ferulic acid-induced insulin secretion in these cells was mediated by augmenting Ca2+ influx through L-type Ca2+ channel. Our data also suggest that this may be a direct, nongenomic action.@*CONCLUSION@#This is the first electrophysiological demonstration that acute ferulic acid treatment could increase L-type Ca2+ channel current in pancreatic β cells by enhancing its voltage dependence of activation, leading to insulin secretion.

Cloning and Expression Patterns of Coumarate-3-hydroxylase Gene from Angelica sinensis and Its Correlation with Content of Ferulic Acid / 中国实验方剂学杂志

ObjectiveTo clone coumarate-3-hydroxylase gene （C3H） from Angelica sinensis， and analyze the correlation between its bioinformatics， expression patterns and content of ferulic acid， and to explore the functions of ASC3H. MethodReal-time polymerase chain reaction （Real-time PCR） was used to clone the full-length cDNA of ASC3H based on the transcriptome dataset of A. sinensis， and the bioinformatics analysis of the gene sequence was carried out. Real-time PCR and high performance liquid chromatography （HPLC） were used to determine relative expression of ASC3H and content of ferulic acid in different root tissues of A. sinensis （periderm， cortex and stele）. ResultThe open reading frame （ORF） of ASC3H （GenBank accession number： MN2550298） was 1 530 bp， encoding 509 amino acids， with a theoretical molecular weight of 57.86 kDa and an isoelectric point of 8.36. It was a hydrophilic protein that was located in the chloroplast with multiple phosphorylation sites and a transmembrane region， and contained a conserved domain CGYDWPKGYGPIINVW_P450 （383-399 aa） in cytochrome P450. Multiple amino acid sequence alignment analysis showed that ASC3H had high similarity with C3H from other plants， especially Ammi majus in Umbelliferae. The Real-time PCR revealed that ASC3H had different expressions in periderm， cortex and stele tissues of A. sinensis roots. It was found from HPLC that the cortex tissues had the highest content of ferulic acid， and the stele tissues had the lowest. ConclusionASC3H was successfully cloned from A. sinensis， and its sequence characteristics were understood more clearly， suggesting that ASC3H might be involved in the ferulic acid biosynthesis pathway of A. sinensis. This paper provided a basis for further studying the functions of the gene and exploring the biosynthesis and regulation mechanism of ferulic acid in A. sinensis， while laying the foundation for the genetic improvement of A. sinensis．

Optimization of the extraction process of Shangke Huoxue Granule by central composite design-response surface methodology / 国际中医中药杂志

Objective:To optimize the extraction process of Shangke Huoxue Granule.Methods:Taking the factors of extraction solvent multiple, extraction time and extraction times as investigation factors, and extraction amount of ferulic acid, paeoniflorin and the ratio of extraction as comprehensive evaluation indices, one-factor experimental design and central composite design-response surface methodology were adopted to optimize the extraction process of Shangke Huoxue Granule.Results:The binomial fitting equation was Y=96.16+2.42 A+0.63 B-3.76 AB-1.57 A2-1.87 B2 ( P<0.01). The optimal extraction process parameters were confirmed to be adding 16 times of water, 64 minutes each time, twice. The deviation rates between the measured values of three verification experiments and the predicted value were 2.00%, 3.23% and 0.66%. Conclusion:The established model of central composite design-response surface methodology has high predictability and the optimized extraction process is stable and feasible.

Methyl ferulic acid ameliorates ethanol-induced L02 cell steatosis through microRNA-378b-mediated CaMKK2-AMPK pathway / 中国中药杂志

Alcoholic liver disease(ALD), with its increasing morbidity and mortality, has seriously and extensively affected the health of people worldwide. Methyl ferulic acid(MFA) has been proven to significantly inhibit alcohol-induced lipid production in L02 cells through the AMP-activated protein kinase(AMPK) pathway, but its in-depth mechanism remains unclear. This study aimed to further clarify the mechanism of MFA in improving lipid accumulation in L02 cells through the microRNA-378b(miR-378b)-mediated calcium/calmodulin-dependent protein kinase kinase 2(CaMKK2)-AMPK signaling pathway based on existing researches. L02 cells were induced by 100 mmol·L~(-1) ethanol for 48 h to establish the model of ALD in vitro, and 100, 50, and 25 μmol·L~(-1) concentration of MFA was treated. MiR-378b plasmids(containing the overexpression plasmid-miR-378b mimics, silence plasmid-miR-378b inhibitor, and their respective negative control-miR-378b NCs) were transfected into L02 cells by electroporation to up-regulate or down-regulate the levels of miR-378b in L02 cells. The levels of total cholesterol(TC) and triglyceride(TG) in cells were detected by commercial diagnostic kits and automatic biochemical analyzers. The expression levels of miR-378b in L02 cells were detected by real-time quantitative polymerase chain reaction(qRT-PCR). CaMKK2 mRNA levels were detected by PCR, and protein expressions of related factors involved in lipid synthesis, decomposition, and transport in lipid metabolism were detected by Western blot. The results displayed that ethanol significantly increased TG and TC levels in L02 cells, while MFA decreased TG and TC levels. Ethanol up-regulated the miR-378b level, while MFA effectively inhibited the miR-378b level. The overexpression of miR-378b led to lipid accumulation in ethanol-induced L02 cells, while the silence of miR-378b improved the lipid deposition induced by ethanol. MFA activated the CaMKK2-AMPK signaling pathway by lowering miR-378b, thus improving lipid synthesis, decomposition, and transport, which improved lipid deposition in L02 cells. This study shows that MFA improves lipid deposition in L02 cells by regulating the CaMKK2-AMPK pathway through miR-378b.

Preservation of phenolic compounds on dried leaf infusion of Bauhinia forficata Link

Abstract Bauhinia forficata Link aqueous extract is usually recommended as a phytomedicine to reduce blood glucose levels and its biological activity has been linked to the presence of phenolic compounds from B. forficata preparations. Several drying processes are used in the production of dry herbal extracts, which may influence the chemical composition and efficacy of final herbal medicines. Due to significant chemical changes, defining appropriate drying processes is essential for phytopharmaceutical drug development. In view of this, we analyzed dried B. forficata leaf infusion (BFLI) extracts by HPLC-UV-MSn, followed by molecular networking analysis to evaluate the chemical profiles from dried extracts yielded by freeze-and spray-drying processes. The main metabolites detected included 11 ferulic/isoferulic acid derivatives and 13 glycosylated flavonoids. The qualitative chemical profiles were alike for both drying processes, whereas the relative abundance of some flavonoids was higher using spray-drying. Taken together, our results showed that freeze-and spray-drying preserved the phenolic profile of BFLI and suggested that spray-drying may be the most suitable to obtain its dried products. Along with studying the chemical profiles of dried herbal extracts, evaluating the influence of drying processes on the quality and chemical profiles of final products is pivotal and may benefit future research.

Chronic Toxicity of Triptolide and Ferulic Acid Ethosomes Gel for Percutaneous Administration / 中国实验方剂学杂志

ObjectiveTo investigate the long-term safety of triptolide ferulic acid ethosome gel in percutaneous administration. MethodWe mixed triptolide with ferulic acid to make liposomes gel in different doses and then administrated the gel to SD rats of both sexes with intact skin and damaged skin for 12 weeks. The daily dosages calculated based on triptolide for the low-， middle-， and high-dose groups were 63.75， 127.50， 255.00 μg·kg-1， respectively. The body weight of each rat was measured weekly. The rats were sacrificed in the last week for the determination of serum biochemical parameters and organ indexes as well as the observation of histopathology. The toxicity was assessed based on the body weight and all the parameters and indexes. ResultAfter long-term administration， the body weight and serum biochemical parameters did not show significant difference between the gel-treated groups and the blank group with intact skin， which indicated that the percutaneous administration of triptolide and ferulic acid ethosomes gel was relatively safe. However， the rats in the high-dose group showed sparse hair and were easy to die in the case of unhairing with chloral hydrate at the late stage of the study. Comprared with the female rats with intact skin in the blank control group， the female rats with damaged skin in the middle-dose group showed decreased heart index （P<0.05）， which indicated certain cardiotoxicity. Moreover， damage appeared in skin and lung， which may be influeneced by dosage， sex， and skin state. ConclusionFerulic acid in combination with triptolide is relatively safe for percutaneous administration， whereas there are some risks of skin and lung damage in the case of long-term administration. Individualized administration scheme should be developed according to liver and kidney function and skin conditons to ensure the safety of clinical medication.

Synthesis of ferulic acid derivatives and their inhibition of cholinesterase inhibitory activity / 中国药科大学学报

@#Ferulic acid (FA), a natural product of phenylpropanoids containing phenolic hydroxyl groups, has a wide range of pharmacological activities and some therapeutic effect on Alzheimer's disease (AD).Using FA as the raw material, the ferulic acid carbamate aniline derivatives were first synthesized by 4-step esterification reaction, splicing carbamate active functional groups, hydrolysis reaction and amide condensation.These FA derivatives were evaluated for in vitro cholinesterase inhibition activity by the Ellman method.A total of 15 novel FA derivatives were designed and synthesized, and their structures were confirmed by 1H NMR, 13C NMR and ESI-MS.Cholinesterase activity tests showed that compounds 5c, 5f, 5j, 5g, 5m possessed good acetylcholinesterase inhibition activity.Except for 5l, 5m, almost all compounds have inhibition activity on butyrylcholinesterase, which is much higher than that on acetylcholinesterase.In conclusion, compounds 5c, 5f, 5j and 5g can be used as potential anti-AD inhibitors targeting cholinesterase..

New and potential properties, characteristics, and analytical methods of ferulic acid: A review

Abstract Phenolic compounds are widely distributed in the plant kingdom and in the microorganisms. Cinnamic acid and its hydroxylated derivative-ferulic acid, are phenolic compounds. Ferulic acid possesses antioxidant potential, as well as anti-cancer, anti-inflammatory, and antimicrobial properties. It prevents the harmful effects of radiation both as an ultraviolet absorber and as a free radical scavenger; it is not cytotoxic. Although ferulic acid has beneficial properties, it is hardly used in cosmetic preparations and has been rarely studied in the literature. Herein, we review the literature on ferulic acid, to provide information which can contribute to further research on the compound.

Effects of Ferulic Acid on the Proliferation ，Invasion and Apoptosis of HepG 2 Hepatocellular Carcinoma Cells / 中国药房

OBJECTIVE：To study the effects of ferulic acid on t he proliferation ，invasion and apoptosis of HepG 2 hepatocelluar carcinoma cells. METHODS ：CCK-8 assay was used to screen the concentration of ferulic acid. Western blot assay was adopted to screen the optimal concentration of interleukin 6（IL-6）to induce HepG 2 cell model with high expression of phosphorylated signal transduction protein and activator 3（p-STAT3）protein. HepG 2 cell were divided into blank control group ， model group ，ferulic acid group （0.5 mmol/L）and positive control group （p-STAT3 inhibitor C 188-9，10 μmol/L）. Except for blank control group ，model group treated with IL- 6，while administration groups were treated with IL- 6 and relevant drugs. Cell survival rate ，invasion and apoptosis rate in early and late stage were detected by CCK- 8 assay，Transwell assay and Annexin V-FITC/PI double staining ，respectively. Western blot assay was used to detect the expression of p-STAT 3，caspase-3，ZBP-89 and vimentin proteins in each group. On the basis of the PDB protein database ，using 1BG1，a highly similar crystal structure of STAT3，as docking template ，using the region around Tyr 705 as the putative binding pocket ，the docking analysis of ferulic acid with STAT 3 protein was carried out. RESULTS ：It is selected to use 0.5 mmol/L ferulic acid intervention for 48 h as the follow-up experimental condition ；50 ng/mL IL- 6 was selected as the modeling condition. Compared with blank control group ，the number of cell invasion ，p-STAT3/STAT3 ratio and protein expression of vimentin were increased significantly in model group （P＜0.05 or P＜0.01），while late apoptosis rate and protein expression 20 of caspase- 3 were decreased significantly （P＜0.05 or P＜ 0.01）. Compared with model group ，cell survival rate ，the number of cell invasion ，p-STAT3/STAT3 ratio and protein expression of vimentin were d ecreased significantly in ferulic acid group and positive control group （P＜0.05 or P＜0.01）；early apoptotic rate （except for ferulic acid group ），late apoptotic rate，the protein expression of caspase- 3 and ZBP- 89（except for positive control group ）were increased significantly （P＜0.05 or P＜0.01）. The results of molecular docking showed that the carboxylic groups of ferulic acid could interact with 1.9 Å hydrogen bond of Asn 581 and 2.0 Å hydrogen bond of Lys 591，with binding energy of －4.4 kcal/mol. CONCLUSIONS ：Ferulic acid may inhibit the activity of p-STAT 3 by directly binding to the phosphorylation site of STAT 3；it may up-regulate the protein expression of caspase- 3 via STAT 3 dependent pathway ，or up-regulate the protein expression of ZBP- 89 via STAT 3 independent pathway and then down-regulate the protein expression of vimentin ，so as to inhibit the proliferation ，invasion and apoptosis of HepG 2 cells.

Anti-tumor activity of rice bran hydrolysates on migration, invasion and angiogenesis

Objective: To investigate anti-tumor effect of rice bran hydrolysates (RBH) on proliferation, migration, invasion, and angiogenesis of cholangiocarcinoma (CCA) cells, and elucidate the underlying mechanisms. Methods: RBH was prepared from Tubtim Chumprae rice (Oryza sativa L.) by hydrothermolysis followed by protease digestion. Phenolic content in RBH was analyzed by high-performance liquid chromatography. Human CCA cells, KKU-156, KKU-452, and KKU-100, were used to study the effects of RBH on proliferation, migration, invasion, and adhesion by wound healing, Transwell chamber, and fibronectin cell adhesion assays. Angiogenesis was evaluated using human umbilical vein endothelial cells. Proteins associated with cancer progression were analyzed by immunobloting assays. Results: RBH contained carbohydrates, proteins, lipids, and various phenolic compounds and flavonoids. RBH did not inhibit CCA proliferation, but strongly suppressed migration, invasion, adhesion of CCA cells, and the formation of tube-like capillary structures of human umbilical vein endothelial cells. Moreover, RBH down-regulated phosphorylation of FAK, PI3K, and Akt, suppressed NF-κB nuclear translocation, decreased the expression of ICAM-1, vimentin and vascular endothelium growth factor (VEGF), and increased the expression of E-cadherin. Conclusions: RBH suppresses CCA cell migration and invasion and decreases expression of proteins involved in cancer metastasis. RBH is a potential food supplement for cancer prevention.

Antibacterial activity and inhibition against Staphylococcus aureus NorA efflux pump by ferulic acid and its esterified derivatives

Objective: To evaluate the inhibitory activity of ferulic acid and four of its esterified derivatives (methyl, ethyl, propyl, and butyl) against resistance mechanisms in Staphylococcus aureus strains. Methods: Ferulic acid derivatives were obtained by esterification with methanol, ethanol, propanol, and butanol, and then characterized by hydrogen and carbon-13 nuclear magnetic resonance analysis. The minimum inhibitory concentrations (MIC) of ferulic acid and its esterified derivatives, ethidium bromide, and norfloxacin were obtained using the microdilution test, while the efflux pump inhibition test was conducted by examining reduction in the MICs of norfloxacin and ethidium bromide. Molecular docking was also carried out using the Schrodinger Suite 2015 molecular modeling software. A three-dimensional model of NorA efflux pump was generated using I-TASSER. The best scoring model was used as a receptor for ligand-receptor docking. Results: The methyl and butyl ester derivatives did not demonstrate significant antimicrobial activity. However, a significant synergic effect was evidenced when norfloxacin was combined with the ethyl and propyl esterified derivatives. The docking study demonstrated favorable energy of interaction between ferulate derivatives and NorA, and amino acid residues TYR57, TYR58, and LEU255 were present commonly in stabilizing all complexes. The PCA analysis corroborated the docking hypothesis that the lipophilic character and hydrogen bond interactions were the most relevant characteristics involved with NorA inhibitors. The pharmacokinetic parameters of ferulic acid derivatives showed good ADMET properties, demonstrating that they can be easily absorbed and have no effect or inhibit the cytochrome P450 enzyme complex, revealing their potential as drug candidates. Conclusions: This study provides strong evidence that the molecular basis for this activity is potentially due to the NorA efflux pump.

Antilipoperoxidative and anti-inflammatory efficacy of multifunctional sunscreens containing ferulic acid

Unprotected chronic exposure to ultraviolet radiation generates many harmful effects to human skin and UV filters are essential to health, however, traditional sunscreens do not provide enough protection against cutaneous oxidative stress, a process amplified by UV radiation. Therefore, is been proposed the development of multifunctional photoprotective formulations, acting in the absorption/reflection of UV radiation and assisting in cutaneous homeostasis. In the present study, ferulic acid is used in conjunction with two sunscreens, bemotrizinol and ethylhexyl triazone, for the determination of biosafety and efficacy methods, using techniques that better elucidate the effects of ferulic acid. Skin permeation assays were performed by applying a formulation containing the three substances in the stratum corneum of volunteers, which were removed by the tape stripping method (ex vivo) with follow quantification by high performance liquid chromatography (HPLC). The test was able to evaluate the penetration depth of the substances, characterizing them. In addition, the simultaneous quantification of the three substances was performed by a single and fast method, facilitating their analysis and improving the technique. Also, TBARS (thiobarbituric acid reactive substances) assays were performed in stratum corneum removed by tape stripping (ex vivo), evaluating the potential of cutaneous lipid peroxidation, with or without ferulic acid. To date, it is the first time that TBARS method is used to characterize the stratum corneum (ex vivo) and quantified by HPLC. The protocol developed may aid in the efficacy of antioxidant agents in studies aimed at elucidating the level of lipid peroxidation caused by drugs and cosmetics, and even in carrying out baseline studies characterizing different ethnicities and genders. As last, an anti-inflammatory in vivo assay with Laser Doppler flowmetry equipment was used to compare the sunscreen formulation with or without ferulic acid. Data indicated that the antioxidant reduced the angular coefficient of the perfusion units, mitigating the inflammatory effects. Furthermore, a significant difference was found between the genders, suggesting a more pronounced inflammatory reaction in women. Ferulic acid proved to be a valuable resource, besides being safe and raise the SPF of sunscreens, it also mitigates the effects of inflammation

A exposição crônica desprotegida à radiação ultravioleta (UV) contribui para o desenvolvimento de câncer de pele e os filtros solares são relevantes para evitar tais efeitos prejudiciais, porém, os protetores solares tradicionais não geram proteção suficiente contra o estresse oxidativo cutâneo. Logo, espera-se o desenvolvimento de formulações fotoprotetoras multifuncionais, atuando não somente na absorção e/ou reflexão da radiação UV, mas, também, auxiliando na homeostase cutânea, com presença de agentes antioxidantes. No presente estudo foi utilizado o ácido ferúlico conjuntamente com dois filtros solares, o bemotrizinol e a triazona de octila, para determinação de métodos de segurança e eficácia, utilizando técnicas que melhor elucidem e comprovem os efeitos do ácido ferúlico. Foram realizados ensaios de permeação cutânea pela aplicação tópica de formulação contendo as três substâncias em voluntários, sendo o estrato córneo retirado pelo método de tape stripping (ex vivo) com subsequente quantificação por cromatografia líquida de alta eficiência (CLAE). O ensaio pôde avaliar a profundidade de penetração das substâncias, caracterizando-as. Ademais, a quantificação simultânea das três substâncias foi efetuada por método único e rápido, facilitando análise com aprimoramento da técnica. Em adição, foi realizado ensaios de TBARS (substâncias reativas ao ácido tiobarbitúrico) em estrato córneo removido por tape stripping (ex vivo), para avaliar o potencial de peroxidação lipídica cutânea, contendo ou não o ácido ferúlico. Até o presente momento, é a primeira vez que o método TBARS é utilizado para caracterização do estrato córneo (ex vivo) e quantificada por CLAE. O protocolo desenvolvido pode auxiliar na eficácia de agentes antioxidantes, em estudos que visam elucidar o nível de peroxidação lipídica causada por medicamentos e cosméticos e, até mesmo, na realização de estudos de base, caracterizando etnias e gêneros. Ademais, um ensaio anti-inflamatório in vivo com equipamento de fluxometria Laser Doppler foi utilizado para comparar a formulação fotoprotetora com ou sem ácido ferúlico. Os dados indicaram que o antioxidante reduziu o coeficiente angular das unidades de perfusão, mitigando os efeitos inflamatórios. Ainda, foi identificada diferença entre os gêneros, sugerindo reação inflamatória mais pronunciada em mulheres. O ácido ferúlico provou ser um recurso valioso, além de ser seguro e elevar o FPS dos fotoprotetores, também atenuando os efeitos da inflamação

Protective Role of Ferulic Acid on Testis-Histoarchitecture and Oxidative Damages Induced by Dimethoate in Rats

Abstract Dimethoate is frequently used all over the world and it caused serious toxicity on target and non-target organisms. In this study, distilled water, ferulic acid, low dose dimetoate, high dose dimetoate, ferulic acid and low dose dimetoate, ferulic acid and high dose dimethoate were given to rats through gavage during the 4-week experiment. For this purpose, the levels of glutathione peroxidase, superoxide dismutase, glutathione-S-transferase, catalase, malondialdehyde and histopathological damages were investigated. After 28 days, no statistically important difference was determined in all investigated parameters in testicular tissues of rats who were administered control and ferulic acid. When the control and ferulic acid groups compare with the low and high dose dimetoate groups, there were statistically significantly changes antioxidant enzymes and malondialdehyde levels. In ferulic acid plus low dose dimethoate treated group and ferulic acid plus high dose dimethoate treatment we demonstrated that the protective role of ferulic acid on examining parameters but not completely. Based on light microscope studies, we can say that both dose dimethoate induced numerous histopathological changes. Less pathological alterations were observed when rats ferulic acid-plus-dimethoate. As a result, it is possible to say that ferulic acid has a partially healing role on the oxidative damage caused by dimethoate.

Ferulic acid alleviates lipopolysaccharide-induced depression-like behavior by inhibiting inflammation and apoptosis

Objective: To identify the anti-depressive effect of ferulic acid (FA) in mice exposed to lipopolysaccharide (LPS) and explore its molecular mechanisms. Methods: The mice were divided into 5 groups as follows: Control, LPS, LPS + SP, LPS + FA, and LPS + FA + anisomycin. The LPS + FA and LPS + FA + anisomycin groups were administered with FA (100 mg/kg, i.p.) once daily continuously for 7 days, and the other groups received an equivalent volume of saline. On the 7th day, LPS (0.1 mg/mL, i.p.) was injected in all mice except the control group 30 min after FA or saline administration. The LPS + SP and LPS + FA + anisomycin groups were intravenously administered with SP600125 [c-Jun N-terminal kinase (JNK) inhibitor] (100 μL/site, i.v.) and anisomycin (JNK activator) (100 μL/site, i.v.) 15 min before LPS, respectively. The depressive behaviors were assessed by open field test, sucrose preference test, and forced swimming test at 24 h post-LPS administration. Tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) levels in plasma were measured by ELISA. The levels of phospho-JNK, TNF-α, IL-1β, Bcl-2, Bax, cytochrome c and caspase-3 were evaluated by Western blotting. Results: FA alleviated depression symptoms caused by LPS in mice, including increasing sucrose water consumption in sucrose preference test and reducing the immobility time in forced swimming test. FA could inhibit upregulated levels of phospho-JNK, TNF-α, and IL-1β. FA also markedly decreased Bax, caspase-3, and cytochrome c, and increased Bcl-2 levels. Besides, SP600125 showed neuroprotective effect similar to FA which was attenuated by anisomycin. Conclusions: FA attenuates inflammation and apoptosis by inhibiting LPS-induced activation of JNK to alleviate depression-like behaviors.

Rice bran hydrolysates induce immunomodulatory effects by suppression of chemotaxis, and modulation of cytokine release and cell-mediated cytotoxicity

Objective: To evaluate the immunomodulatory effects of rice bran hydrolysates on cultured immune cells and their underlying mechanism. Methods: Rice bran hydrolysates were prepared from pigmented rice (Oryza sativa L.) by hydrothermolysis and protease digestion. Rice bran hydrolysates were assayed for phenolic content and antioxidant activity. Cell proliferation of Jurkat, THP-1 and peripheral blood mononuclear cells (PBMC) was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Chemotaxis was evaluated by transwell chamber methods. Immunoadherence of THP-1 was performed on cultured human umbilical vein endothelial cells (HUVEC). Cytokine released from PBMC was measured by ELISA assay kits. Lymphocyte-mediated cytotoxicity was carried out on KKU-452 cells. Proteins associated with immunomodulation were analyzed by Western immunoblotting assay. Results: Rice bran hydrolysates were rich in phenolic compounds, such as ferulic acid, catechin, quercetin, and quercetin glycosides. Rice bran hydrolysates suppressed phytohemagglutinin (PHA)-stimulated proliferation of PBMC and Jurkat cells, chemotaxis of Jurkat and THP-1 cells, and immunoadherence of THP-1 on HUVEC cultured cells. The cellular mechanism of rice bran hydrolysates involved the activation of AMPK as well as suppression of mTOR, NF-κB and VCAM-1. Rice bran hydrolysates potentiated PBMC on the PHA-stimulated release of IL-2, TNF-α, and IL-4, and enhanced PHA-induced non-MHC-restricted cytotoxicity on KKU-452 cancer cells. Conclusions: The immunomodulatory effect of phytochemicals derived from rice bran hydrolysates suggests its therapeutic potential for further investigation.

Orthogonal compatibility study on main effective parts of Yangyin Tongnao Granules based on pharmacokinetics and pharmacodynamics / 中草药

Objective: To investigate the pharmacokinetic and pharmacodynamic changes of Yangyin Tongnao Granules (YTG) in cerebral ischemia reperfusion rats after the compatibility of main effective parts (total alkaloids, total flavonoids, total saponins and total phenolic acids). Methods: By using the orthogonal design to research the main effective parts of YTG, nine different dosage ratios combinations were formed, which were used for oral administration in cerebral ischemia reperfusion rats. High performance liquid chromatography-diode array detector (HPLC-DAD) was used to determine the concentration of puerarin, ferulic acid, and ligustrazine in rat plasma at different time points. The non-compartmental model was used to fit the pharmacokinetic parameters by Drug and Statistics (DAS) 3.2.6 software. The total quantum statistical moment analysis method and comprehensive evaluation method were used to evaluate the total pharmacokinetic characteristics. Meanwhile, the content of superoxide dismutase (SOD) and catalase (CAT) were determined by enzyme-linked immunosorbent assay (ELISA) kits. Finally, the PK-PD model and the quantitative equations between drug concentration and efficacy were obtained. Results: The pharmacokinetic characteristics of puerarin, ferulic acid and ligustrazine in cerebral ischemia-reperfusion rats were different. Total statistical moment and comprehensive score study showed that different combinations had different effects on ACUT, mean retention time (MRT), and comprehensive evaluation. The effective parts inhibited the reduction of oxidation indexes such as SOD and CAT. Sigmoid-Emax models were adopted in all PK-PD models, and the fitting results had a good correlation with the measured data. The R values were more than 0.85. Conclusion: Compatibility of YTG activity parts had a certain effect on their pharmacokinetic behaviors and antioxidation in model rats. The total quantum statistical moment analysis and comprehensive evaluation method can be used to study the pharmacokinetics of multi-component traditional Chinese medicine prescriptions. PK-PD model could be used to predict and evaluate the correlation between pharmacokinetics and pharmacodynamics of traditional Chinese medicine prescriptions.

Selection of high altitude planting area of Angelica sinensis based on biomass, bioactive compounds accumulation and antioxidant capacity / 中草药

Objective: To select the high altitude planting area for improving the quality of Angelica sinensis based on biomass, bioactive compounds accumulation and antioxidant capacity in rhizome. Methods: The experiments were conducted on dry weight (DW), bioactive compounds content and antioxidant capacity in two-year old rhizome of A. sinensis grown at different altitudes (2 100, 2 500, and 2 900 m), under controlling plants reproduced from the seeds of the same plant, the same levels of water and fertilizer in soil. Results: Higher altitude was adverse to rhizome biomass, however, the compounds content (DW and per plant basis) and antioxidant capacity were significantly promoted at higher altitude compared to lower altitude site; On a per plant basis, the content of ferulic acid, soluble sugar, phenolics and flavonoids in rhizome of un-bolted plants grown at 2 900 m respectively was 2.06, 1.13, 1.34 and 1.15 fold greater than that of 2 100 m; The antioxidant capacity also increased with higher altitude. Comprehensive analysis showed that total content of main bioactive compounds in rhizome of un-bolted plants increased with higher altitudes ranging from 2 100 to 2 900 m. Conclusion: Higher-altitude cultivation can significantly enhance bioactive compounds accumulation in rhizome. The biosynthesis and accumulation of bioactive compounds were regulated by low temperature and light. These findings will provide theoretical references for improving production and quality of rhizome as well as large-scale cultivation.

Chemical constituents and cytotoxic activities of Dendrobium wardianum / 中草药

Objective To study the chemical constituents and cytotoxic activity of Dendrobium wardianum. Methods The compounds were separated and purified by various column chromatography techniques and semi-preparative liquid chromatography, and the structures were then identified by NMR, MS and TLC. And the cytotoxic activity of compounds 3, 6, 8, 9, 11, 14, 15 were detected by MTS assay. Results Sixteen compounds were isolated from D. wardianum. They were identified as amotin (1), aduncin (2), E-ferulic acid eicosyl ester (3), E-ferulic acid docosyl ester (4), tetracosyl p-coumarate (5), 4-hydroxy-3-methoxy cinnamaldehyde (6), dihydroconiferyl alcohol (7), 3,7-dimethoxy-5-hydroxy-1,4-phenanthrenequinone (8), 3-ethoxy-5-hydroxy-7- methoxy-1,4-phenanthrenequinone (9), 4-methoxy-9,10-dihydrophilic-2,7-diol (10), 5,7,4’-trihydroxy dihydroflavone (11), 5,7,4’- trihydroxy-3’,5’-dimethoxy dihydroflavone (12), tricin (13), 3,5,4’-trimethoxy-4-hydroxybibenzyl (14), 3,5,3’-trimethoxy-4’- hydroxybibenzyl (15) and 3,3’-dimethoxy-4,5’-dihydroxybibenzyl (16). Compounds 6 and 8 have tumor growth inhibitory activity on tumor growth in vitro. Conclusion Compounds 3, 5, 6 are isolated from Dendrobium for the first time, and 10, 11, 14, 15 are isolated from D. wardianum for the first time. The active components of D. wardianum need to be further explored.

Simultaneous determination of 13 bioactive components in Jiawei Zuojin Pill by HPLC-MS/MS / 中草药

Objective: To develop and validate an high performance liquid chromatography coupled with electrospray tandem mass spectrometry (HPLC-ESI-MS/MS) method for simultaneously qualitative and quantitative determination of 13 major bioactive components (ferulic acid, costunolide, baicalin, paeoniflorin, tetrahydropalmatine, rutecarpine, berberine, palmatine, evodiamine, naringin, hesperidin, saikosaponin a, and saikosaponin d) in Jiawei Zuojin Pills (JZP). Methods: The chromatographic separation was performed on a Thermo Syncronis C18 column (100 mm × 4.6 mm, 3.0 μm) with a gradient elution of methanol and 0.02 mol/L ammonium acetate in water at a flow rate of 0.35 mL/min, and the injection volume was 10 μL. The 13 major bioactive components were detected using an electrospray ionization source in ESI+ and ESI- ionization mode, quantified by multiple reaction monitor (MRM) scanning at the same time. Results: The linear ranges of ferulic acid, costunolide, baicalin, paeoniflorin, tetrahydropalmatine, rutecarpine, berberine, palmatine, evodiamine, naringin, hesperidin, saikosaponin a and saikosaponin d were 2-80 μg/mL (r = 0.999 2), 0.5-20.0 μg/mL (r = 0.999 1), 3.5-140.0 μg/mL (r = 0.999 8), 1-40 μg/mL (r = 0.999 2), 0.3-12.0 μg/mL (r = 0.999 1), 1-40 μg/mL (r = 0.999 2), 3-120 μg/mL (r = 0.999 7), 2.5-100.0 μg/mL (r = 0.999 5), 0.5-20.0 μg/mL (r = 0.999 3), 0.5-20.0 μg/mL (r = 0.999 1), 1-40 μg/mL (r = 0.999 1), 0.3-12.0 μg/mL (r = 0.999 2), 0.3-12.0 μg/mL (r = 0.999 2), and the average recoveries were 99.5% (RSD = 4.11%), 98.9% (RSD = 4.88%), 100.2% (RSD = 1.08%), 99.2% (RSD = 3.23%), 99.5% (RSD = 4.13%), 99.7% (RSD = 3.23%), 98.6% (RSD = 2.78%), 99.9% (RSD = 3.12%), 101.3% (RSD = 4.53%), 98.7% (RSD = 3.43%), 99.8% (RSD = 3.58%), 97.9% (RSD = 5.22%), and 101.3% (RSD = 5.13%), respectively. The contents of 12 batches of the 13 major bioactive components were 0.324-0.383, 0.051-0.072, 3.225-3.466, 0.154-0.198, 0.015-0.062, 0.144-0.199, 2.145-2.982, 0.441-0.953, 0.032-0.099, 0.062-0.089, 0.111-0.178, 0.012-0.065, 0.011-0.069 mg/g, respectively. Conclusion: The developed method is simple, specific, and sensitive, and it can be applied for the determination of 13 major bioactive components and the quality control of JZP.