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.
Rats ; Animals ; Insulin Secretion ; Insulin/pharmacology* ; Insulin-Secreting Cells/metabolism* ; Coumaric Acids/metabolism* ; Calcium/metabolism*

A field trial was carried out to study the influence of different kinds of spring topdressing on growth, yield and quality of Ligusticum chuanxiong. The results showed that the spring topdressing had effects of improving root length, tiller numbers and plant height to some extent. At the same time the chlorophyll content and dry weight accumulation especially the dry weight of root increased significantly. It also showed that the yield increased and quality was improved significantly. The effect of different treatment with urea58.7 kg x hm(-2)(N 27 kg x hm(-2)) was the best and the treatment with N,P,K the second.
Alkaloids ; metabolism ; Coumaric Acids ; metabolism ; Fertilizers ; Ligusticum ; growth & development ; metabolism ; Seasons

A mathematical model of microwave drying kinetics based on Fick's second diffusion law was established and and the influence of microwave drying on ferulic acid (FA) and hydrosafflower yellow A (HSYA) in Shuxiong prescription was studied. The proposed model has been verified by using experimental data from microwave drying of Shuxiong prescription. By increasing the microwave output power (500-650 W) and the sample amount (15-60 g), the diversion rate of FA and HSYA changed. In order to determine the kinetic parameters, the drying data were fifed to various models based on the ratios of differences between the initial and final moisture contents and equilibrium moisture content versus drying time. The activation energy was calculated using an exponential expression based on Arrhenius equation. The relationship between the drying rate constant and effective moisture diffusivity was also estimated, and it showed linear.
Coumaric Acids ; metabolism ; Desiccation ; methods ; Diffusion ; Drugs, Chinese Herbal ; chemistry ; metabolism ; Kinetics ; Microwaves ; Models, Theoretical ; Pharmaceutical Preparations ; chemistry ; metabolism

Ferulic acid (FA) is an active component of herbal medicines. One of the best documented activities of FA is its antioxidant property. Moreover, FA exerts antiallergic, anti-inflammatory, and hepatoprotective effects. However, the metabolic pathways of FA in humans remain unclear. To identify whether human CYP or UGT enzymes are involved in the metabolism of FA, reaction phenotyping of FA was conducted using major CYP-selective chemical inhibitors together with individual CYP and UGT Supersomes. The CYP- and/or UGT-mediated metabolism kinetics were examined simultaneously or individually. Relative activity factor and total normalized rate approaches were used to assess the relative contributions of each major human CYPs towards the FA metabolism. Incubations of FA with human liver microsomes (HLM) displayed NADPH- and UDPGA-dependent metabolism with multiple CYP and UGT isoforms involved. CYPs and UGTs contributed equally to the metabolism of FA in HLM. Although CYP1A2 and CYP3A4 appeared to be the major contributors in the CYP-mediated clearance, their contributions to the overall clearance are still minor (< 25%). As a constitute of many food and herbs, FA poses low drug-drug interaction risk when co-administrated with other herbs or conventional medicines because multiple phase I and phase II enzymes are involved in its metabolism.
Coumaric Acids ; chemistry ; metabolism ; Cytochrome P-450 Enzyme System ; chemistry ; metabolism ; Drugs, Chinese Herbal ; metabolism ; Glucuronosyltransferase ; chemistry ; metabolism ; Humans ; Kinetics ; Medicine, Chinese Traditional ; Microsomes, Liver ; chemistry ; enzymology

Objective: To investigate the regulatory effects and signaling mechanism of sodium ferulate on the proliferation and apoptosis of human skin hypertrophic scar fibroblasts (HSFbs). Methods: The experimental research methods were used. The 4^th-6^th passage of HSFbs from human skin were used for the following experiments. HSFbs were co-cultured with sodium ferulate at final mass concentrations of 1, 1×10^-1, 1×10^-2, 1×10^-3, 1×10^-4, 1×10^-5, and 1×10^-6 mg/mL for 48 hours, and methyl thiazolyl tetrazolium method was used to determine the cell absorbance values and linear regression was used to analyze the half lethal concentration (LC50) of sodium ferulate (n=6). HSFbs were co-cultured with sodium ferulate at final mass concentrations of 0.1, 0.2, 0.3, and 0.4 mg/mL for 24, 48, 72, and 96 hours, and methyl thiazolyl tetrazolium method was used to determine the cell absorbance values and the cell proliferation inhibition rate was calculated (n=3). According to the random number table, the cells were divided into 0.300 mg/mL sodium ferulate group, 0.030 mg/mL sodium ferulate group, 0.003 mg/mL sodium ferulate group treated with sodium ferulate at corresponding final mass concentrations, and negative control group without any treatment. After 72 hours of culture, the cell absorbance values were determined by methyl thiazolyl tetrazolium method (n=5), the microscopic morphology of cells was observed by transmission electron microscope (n=3), the cell apoptosis was detected by TdT-mediated dUTP-biotin nick end labeling (TUNEL) assay and the apoptosis index was calculated (n=4), the protein expressions of B lymphocystoma-2 (Bcl-2), Bcl-2-associated X protein (Bax), and cysteine aspartic acid specific protease-3 (caspase-3) were determined by immunohistochemistry (n=4), and the protein expressions of transformed growth factor β₁ (TGF-β₁), phosphorylated Smad2/3, phosphorylated Smad4, and phosphorylated Smad7 were detected by Western blotting (n=4). Data were statistically analyzed with one-way analysis of variance and Dunnett test. Results: The LC50 of sodium ferulate was 0.307 5 mg/mL. After being cultured for 24-96 hours, the cell proliferation inhibition rates of cells treated with sodium ferulate at four different mass concentrations tended to increase at first but decrease later, which reached the highest after 72 hours of culture, so 72 hours was chosen as the processing time for the subsequent experiments. After 72 hours of culture, the cell absorbance values in 0.003 mg/mL sodium ferulate group, 0.030 mg/mL sodium ferulate group, and 0.300 mg/mL sodium ferulate group were 0.57±0.06, 0.53±0.04, 0.45±0.05, respectively, which were significantly lower than 0.69±0.06 in negative control group (P<0.01). After 72 hours of culture, compared with those in negative control group, the cells in the three groups treated with sodium ferulate showed varying degrees of nuclear pyknosis, fracture, or lysis, and chromatin loss. In the cytoplasm, mitochondria were swollen, the rough endoplasmic reticulum was expanded, and local vacuolation gradually appeared. After 72 hours of culture, compared with that in negative control group, the apoptosis indexes of cells were increased significantly in 0.003 mg/mL sodium ferulate group, 0.030 mg/mL sodium ferulate group, and 0.300 mg/mL sodium ferulate group (P<0.05 or P<0.01). After 72 hours of culture, compared with those in negative control group, the protein expressions of Bcl-2 of cells in 0.300 mg/mL sodium ferulate group was significantly decreased (P<0.01), the protein expressions of Bax of cells in 0.030 mg/mL sodium ferulate group and 0.300 mg/mL sodium ferulate group were significantly increased (P<0.05), and the protein expression of caspase-3 of cells in 0.300 mg/mL sodium ferulate group was significantly increased (P<0.01). After 72 hours of culture, compared with those in negative control group, the protein expression levels of TGF-β₁, phosphorylated Smad2/3, and phosphorylated Smad4 of cells in 0.030 mg/mL sodium ferulate group and 0.300 mg/mL sodium ferulate group were significantly decreased (P<0.05 or P<0.01), and the protein expression levels of phosphorylated Smad7 of cells in 0.003 mg/mL sodium ferulate group, 0.030 mg/mL sodium ferulate group, and 0.300 mg/mL sodium ferulate group were significantly increased (P<0.01). Conclusions: Sodium ferulate can inhibit the proliferation of HSFbs of human skin and promote the apoptosis of HSFbs of human skin by blocking the expression of key proteins on the TGF-β/Smad signaling pathway and synergistically activating the mitochon- drial apoptosis pathway.
Apoptosis ; Caspase 3/metabolism* ; Cell Proliferation ; Cicatrix, Hypertrophic/metabolism* ; Coumaric Acids ; Fibroblasts/metabolism* ; Humans ; Signal Transduction ; bcl-2-Associated X Protein/pharmacology*

OBJECTIVESome of the phenolic compounds detected in the soil of commercially cultivated American ginseng could inhibit the seed germination and seedling growth of American ginseng. In this paper we studied the root exudation of American ginseng induced by deficiency of nitrogen, phosphorus and potassium on the content of phenolic compounds.

METHODTwo years old American ginsengs were cultured in hydroponic culture with different nutrient solution. The culture solution was collected after 14 days. The exudations of different polarities in the culture solution were enriched by the amberlite XAD4 and XAD7. The content of the total phenolic acids in the exudation was analyzed by Folin-Ciocalteu colorimetry; the contents of vanillic acid, p-coumaric acid and trans-cinnamic acid were detected and quantified by HPLC.

RESULTBoth in the situation of nitrogen and potassium deficiency, the concentration of total phenolic compounds increased significantly in the exudation of American ginseng comparing with the complete nutrient solution (P < 0.05) , while decreased significantly under phosphorus deficient conditions (P < 0.05). The contents of the 3 autotoxic phenolic acids decreased significantly under nitrogen, phosphorus and potassium deficient conditions (P < 0.05).

CONCLUSIONThe contents of total phenolic compounds and the 3 autotoxic phenolics in the root exudation of American ginseng altered variously in the deficiency of nitrogen, phosphorus and potassium.

Coumaric Acids ; analysis ; Hydroxybenzoates ; analysis ; Nitrogen ; deficiency ; Panax ; chemistry ; metabolism ; Phosphorus ; deficiency ; Plant Exudates ; chemistry ; Plant Roots ; chemistry ; metabolism ; Potassium ; chemistry ; Propionates ; Soil ; chemistry ; Vanillic Acid ; analysis

AIMTo study the effects of ferulic acid (FA) on E-selectin expression in human umbilical vein endothelial cells (HUVECs) activated by lipopolysaccharide and leukocyte-endothelial cell adhesion.

METHODSThe effects of FA on E-selectin and E-selectin mRNA expression were determined by flow cytometry and reverse transcription polymerase chain reaction. The effect of FA on HL60-HUVEC adhesion was evaluated with the method of staining the cells by Rose Bengal.

RESULTSThe expression of E-selectin and E-selectin mRNA were down regulated by FA (0.62 and 0.41 mmol x L(-1), respectively). HL60 cells adhered to activated HUVECs were also reduced by FA (0.62 and 0.41 mmol x L(-1), respectively).

CONCLUSIONFA can inhibit the expression of E-selectin and E-selectin mRNA and HL60-HUVEC adhesion. This may contribute to its protective effect against ischemia-reperfusion injury.

Cell Adhesion ; drug effects ; Cells, Cultured ; Coumaric Acids ; pharmacology ; E-Selectin ; biosynthesis ; genetics ; Endothelial Cells ; metabolism ; HL-60 Cells ; physiology ; Humans ; RNA, Messenger ; biosynthesis ; genetics ; Umbilical Veins ; cytology

To establish a method for detecting microdialysis recovery of tetramethylpyrazine (TMP) and ferulic acid (FA) and investigating the influencing factors, providing the basis for further in vivo microdialysis experiments. The concentration of FA and TMP in dialysates were determined by high pressure liquid chromatography ( HPLC) and probe recovery were calculated respectively. The influence of the flow rates, medium concentration, temperature and in vivo probe stability on the recovery of FA and TMP were investigated by using concentration difference method (incremental method and decrement method). The recovery obtained by incremental method were similar to by decrement method. The in vitro recovery rate of FA and TMP decreased with the increase of 1-2.5 μL min(-1), and increased obviously with the temperature of 25-42 degrees C under the same conditions. The concentration of FA and TMP had no obvious effect on the probe recovery under the same flow rate. In addition, the recovery of TMP and FA remained stable and showed similar trends under the condition of four concentration cycles, indicating that the intra day reproducibility of the concentration difference method was good. The recovery of brain microdialysis probes in vivo 8 h maintained a relatively stable, but certain differences existed between different brain microdialysis probes, demonstrating that each probe was required for recovery correction in vivo experiment. Microdialysis sampling can be used for the local brain pharmacokinetic study of FA and TMP, and retrodialysis method can be used in probe recovery of FA and TMP in vivo.
Animals ; Brain ; metabolism ; Chromatography, High Pressure Liquid ; Coumaric Acids ; analysis ; isolation & purification ; pharmacokinetics ; Drugs, Chinese Herbal ; Humans ; Microdialysis ; methods ; Pyrazines ; analysis ; isolation & purification ; pharmacokinetics ; Rats

To express feruloyl esterase A from Aspergillus oryzae in Pichia pastoris expression system and study its hydrolysis function, explore the conditions and effects of purification for ferulic acid extracts by macroporos resin. Using the total RNA from A. oryzae CICC 40186 as the template, we amplified coding sequence AorfaeA encoding a mature feruloyl esterase A (AorFaeA) by RT-PCR technique. Then, the coding sequence AorfaeA was successfully expressed in Pichia pastoris GS115 mediated by an expression plasmid pPIC9K. The purified recombinant AorFaeA (reAorFaeA) showed one single band on SDS-PAGE with an apparent molecular weight of 39.0 kDa. The maximum activity of reAorFaeA to methyl ferulate, measured by high-performance liquid chromatography (HPLC), was 58.35 U/mg. Then, reAorFaeA was used to release ferulic acid from de-starched wheat bran in the presence of xylanase. The purification tests for ferulic acid from the enzymatic hydrolysate were carried out with preselected macroporous resins. The results showed that macroporous resin HPD-300 had much higher adsorption and desorption capacities. Ferulic acid could be quantitatively recovered by 50% of the eluent concentration at a flow speed of 1 mL/min. Under the purification condition, the recovery ratio of ferulic acid was 92%, and the content of ferulic acid was increased from 0.13% in the raw material to 10.55%. This work exploits the breakdown of ferulic acid by recombinant enzymeand provids a good strategy to its "green production".
Aspergillus oryzae ; enzymology ; Carboxylic Ester Hydrolases ; biosynthesis ; genetics ; Cloning, Molecular ; Coumaric Acids ; chemistry ; Electrophoresis, Polyacrylamide Gel ; Hydrolysis ; Molecular Weight ; Pichia ; genetics ; metabolism

AIMTo investigate the effect of sodium ferulate (SF) on glycerol-induced renal injury.

METHODSGlycerol solution 50% was injected intramuscularly to establish a model of acute tubular necrosis in mice. SF was administered intraperitoneally at the dose of 100-200 mg.kg-1 at the beginning of establishing the model and its effect was observed by monitoring renal function, antioxidative functions and renal pathologic histology.

RESULTSAt 6 and 72 h after glycerol injection, SF treatment (100-200 mg.kg-1) showed significant and dose-dependent antagonistic actions on the increment of blood urea nitrogen (BUN), creatinine (Cr), and N-acetyl-beta-glucosaminidase (NAG) induced by glycerol. The increase of renal malondialdehyde (MDA) content and the decrease of glutathione content, glutathione peroxidase (GSH-Px), glutathione S-transferase (GST), catalase (Cat) and superoxide dismutase (SOD) activities resulting from glycerol injection were remarkably inversed by SF at the dose of 200 mg.kg-1. Meanwhile, improvement of the renal histology was observed as well.

CONCLUSIONSF showed beneficial effect on glycerol-induced acute tubular necrosis due to its antioxidative action.

Animals ; Antioxidants ; pharmacology ; Blood Urea Nitrogen ; Coumaric Acids ; pharmacology ; Creatinine ; blood ; Glutathione Peroxidase ; metabolism ; Glycerol ; Kidney ; metabolism ; pathology ; Kidney Function Tests ; Kidney Tubular Necrosis, Acute ; chemically induced ; metabolism ; pathology ; Male ; Malondialdehyde ; metabolism ; Mice ; Superoxide Dismutase ; metabolism