Neuroprotective Effects of P-Coumaric Acid on Haloperidol-Induced Catalepsy Through Ameliorating Oxidative Stress and Brain Dopamine Level

Objective To evaluate the effect of p-coumaric acid (p-CA) on haloperidol-induced catalepsy in Swiss albino male mice. Method To induce catalepsy, haloperidol (1 mg/kg i.p.) was administered for 21 consecutive days. p-CA (50, 75, and 100 mg/kg, PO) was administered 30 min before haloperidol injection for 21 consecutive days. For catalepsy, locomotor activity and motor coordination scores were recorded on the 17, 14, and 21 days of drug treatment, while the gait analysis score was recorded on day 21. After behavioral testing, animals were sacrificed, and various biochemical and histopathology tests of the brain were conducted. Dopamine, malondialdehyde, reduced glutathione (GSH), superoxide dismutase (SOD), and catalase activity were examined in the brain. Results Chronic administration of haloperidol significantly increased catalepsy in mice. It also produced hypolocomotion, motor coordination, and gait disturbance in mice. p-CA significantly inhibited haloperidol-induced catalepsy. Haloperidol significantly increased malondialdehyde levels in the brain. While dopamine levels in the brain dropped along with GSH, SOD, and catalase activity levels, which also had an impact on the histology of the brain. p-CA significantly reduced haloperidol-induced increases in brain oxidative stress, dopamine levels in the brain, and brain histology in mice. Discussion p-CA significantly reduced haloperidol-induced catalepsy, possibly through reducing oxidative stress and increasing brain dopamine levels. It can be a good candidate drug for extrapyramidal symptoms in Parkinson’s disease and adjuvant therapy with antipsychotic drugs.

Effect of P-coumaric Acid on Apoptosis of Multiple Myeloma Cells Based on Oxidative Stress / 中国实验血液学杂志

OBJECTIVE@#To investigate the effect of p-coumaric acid on apoptosis of multiple myeloma cells and its related mechanism.@*METHODS@#Multiple myeloma cell line MM.1s cells were selected and treated with different concentrations of p-coumaric acid (0, 0.4, 0.8, 1.6, 3.2 mmol/L), and the inhibition rate and half inhibition concentration (IC50) were detected by CCK-8 method. Then MM.1s cells were treated with 1/2 IC50, IC50, 2 IC50 and transfected with ov-Nrf-2 and ov-Nrf-2+IC50. The apoptosis, ROS fluorescence intensity and mitochondrial membrane potential of MM.1s cells were detected by flow cytometry, and the relative expressions of cellular Nrf-2 and HO-1 protein were detected by Western blot.@*RESULTS@#P-coumaric acid inhibited the proliferation of MM.1s cells in a dose-dependent manner(r =0.997) with an IC50 value of 2.754 mmol/L. Compared with the control group, apoptosis and ROS fluorescence intensity of MM.1s cells were significantly increased in the 1/2 IC50 group, IC50 group, 2 IC50 group and ov-Nrf-2+IC50 group (P <0.01), the expressions of Nrf-2, HO-1 protein in the IC50 group and 2 IC50 group were significantly decreased (P <0.05). Compared with the IC50 group, the cells apoptosis and ROS fluorescence intensity were significantly decreased (P <0.01), and the expressions of Nrf-2 and HO-1 protein were significantly increased in the ov-Nrf-2+IC50 group (P <0.01).@*CONCLUSION@#P-coumaric acid can inhibit the proliferation of MM.1s cells and may target the Nrf-2/HO-1 signaling pathway to affect oxidative stress in MM cells thereby inducing their apoptosis.

Characterization of highly active tyrosine ammonia lyase and its application in biosynthesis of p-coumaric acid / 生物工程学报

p-coumaric acid is one of the aromatic compounds that are widely used in food, cosmetics and medicine due to its properties of antibacterium, antioxidation and cardiovascular disease prevention. Tyrosine ammonia-lyase (TAL) catalyzes the deamination of tyrosine to p-coumaric acid. However, the lack of highly active and specific tyrosine ammonia lyase limits cost-effective microbial production of p-coumaric acid. In order to improve biosynthesis efficiency of p-coumaric acid, two tyrosine ammonia-lyases, namely Fc-TAL2 derived from Flavobacterium columnare and Fs-TAL derived from Flavobacterium suncheonense, were selected and characterized. The optimum temperature (55 ℃) and pH (9.5) for Fs-TAL and Fc-TAL2 are the same. Under optimal conditions, the specific enzyme activity of Fs-TAL and Fc-TAL2 were 82.47 U/mg and 13.27 U/mg, respectively. Structural simulation and alignment analysis showed that the orientation of the phenolic hydroxyl group of the conserved Y50 residue on the inner lid loop and its distance to the substrate were the main reasons accounting for the higher activity of Fs-TAL than that of Fc-TAL2. The higher activity and specificity of Fs-TAL were further confirmed via whole-cell catalysis using recombinant Escherichia coli, which could convert 10 g/L tyrosine into 6.2 g/L p-coumaric acid with a yield of 67.9%. This study provides alternative tyrosine ammonia-lyases and may facilitate the microbial production of p-coumaric acid and its derivatives.

Effect of key enzymes ubiquitination sites on the biosynthesis of naringenin / 生物工程学报

Flavonoids have a variety of biological activities and have important applications in food, medicine, cosmetics, and many other fields. Naringenin is a platform chemical for the biosynthesis of many important flavonoids. Ubiquitination plays a pivotal role in the post-translational modification of proteins and participates in the regulation of cellular activities. Ubiquitinated proteins can be degraded by the ubiquitin-protease system, which is important for maintaining the physiological activities of cells, and may also exert a significant impact on the expression of exogenous proteins. In this study, a real-time in-situ detection system for ubiquitination modification has been established in Saccharomyces cerevisiae by using a fluorescence bimolecular complementation approach. The ubiquitination level of protein was characterized by fluorescence intensity. By using the approach, the potential ubiquitination sites of proteins involved in the naringenin biosynthesis pathway have been obtained. The lysine residues of the relevant ubiquitination sites were mutated to arginine to reduce the ubiquitination level. The mutants of tyrosine ammonia-lyase (FjTAL) and chalcone synthase (SjCHS, SmCHS) showed decreased fluorescence, suggested that a decreased ubiquitination level. After fermentation verification, the S. cerevisiae expressing tyrosine ammonia-lyase FjTAL mutant FjTAL-K487R accumulated 74.2 mg/L p-coumaric acid at 72 h, which was 32.3% higher than that of the original FjTAL. The strains expressing chalcone synthase mutants showed no significant change in the titer of naringenin. The results showed that mutation of the potential ubiquitination sites of proteins involved in the naringenin biosynthesis pathway could increase the titer of p-coumaric acid and have positive effect on naringenin biosynthesis.

Determination of six effective ingredients in Imperatae Rhizoma concentrated granules by ultra-performance liquid chromatography using two internal references / 药学学报

To simultaneously determine the contents of p-coumaric acid, chlorogenic acid, 5-caffeoylquinic acid, 4-caffeoylquinic acid, caffeic acid and ferulic acid in Imperatae Rhizoma concentrated granules, an ultra-high performance liquid chromatography (UPLC) with two internal references method (TIRM) was established and validated. Chromatographic separation was achieved on a ZORBAX RRHD Eclipse Plus C18 column (2.1 mm × 100 mm, 1.8 µm) using 1.7 mmol·L-1 oxalic acid in water and methanol as mobile phase. The flow rate was 0.4 mL·min-1 and the column temperature was set as 35 ℃. The relative correction factors (RCFs) of caffeic acid and ferulic acid using p-coumaric acid as internal reference were calculated and the RCFs of 4-caffeoylquinic acid and 5-caffeoylquinic acid were calculated using chlorogenic acid as the internal reference. The TIRM was fully validated for linearity, accuracy, repeatability, stability and recovery so that it could be compared with the external standard method (ESM). The RCFs of 5-caffeoylquinic acid, 4-caffeoylquinic acid, caffeic acid, and ferulic acid were 1.069, 1.022, 1.368, and 1.493, respectively. The TIRM and ESM were used to determine the contents of six ingredients in Imperatae Rhizoma concentrated granules from different manufacturers and the variation between results was within acceptable limits. In conclusion, the newly established TIRM allowed simultaneous determination of six ingredients (p-coumaric acid, chlorogenic acid, 5-caffeoylquinic acid, 4-caffeoylquinic acid, caffeic acid, ferulic acid) in Imperatae Rhizoma concentrated granules, providing support for the quality control of this traditional Chinese medicine.

High-throughput screening of Saccharomyces cerevisiae efficiently producing tyrosine / 生物工程学报

Tyrosine is an important aromatic amino acid. Besides its nutritional value, tyrosine is also an important precursor for the synthesis of coumarins and flavonoids. Previously, our laboratory constructed a Saccharomyces cerevisiae strain LTH0 (ARO4K229L, ARO7G141S, Δaro10, Δzwf1, Δura3) where tyrosine feedback inhibition was released. In the present study, heterologous expression of betaxanthins synthesis genes DOD (from Mirabilis jalapa) and CYP76AD1 (from sugar beet B. vulgaris) in strain LTH0 enabled production of yellow fluorescence. The engineered strain LTH0-DOD-CYP76AD1 was subjected to UV combined with ARTP mutagenesis, followed by flow cytometry screening. Among the mutants screened, the fluorescence intensity of the mutant strain LTH2-5-DOD-CYP76AD1 at the excitation wavelength of 485 nm and emission wavelength of 505 nm was (5 941±435) AU/OD, which was 8.37 times higher than that of strain LTH0-DOD-CYP76AD1. Fourteen mutant strains were subjected to fermentation to evaluate their tyrosine producing ability. The highest extracellular tyrosine titer reached 26.8 mg/L, which was 3.96 times higher than that of strain LTH0-DOD-CYP76AD1. Heterologous expression of the tyrosine ammonia lyase FjTAL derived from Flavobacterium johnsoniae further increased the titer of coumaric acid to 119.8 mg/L, which was 1.02 times higher than that of the original strain LTH0-FjTAL.

p-Coumaric acid alleviates adriamycin-induced hepatotoxicity in rats

Objective: To evaluate the effect of p-coumaric acid against adriamycin-induced hepatotoxicity in rats. Methods: The rats were divided into 4 groups. The control group received solvent; the p-coumaric acid group was treated with 100 mg/kg of p-coumaric acid orally for five consecutive days; the adriamycin group was administered with a single dose of adriamycin (15 mg/kg, i.p.), and the p-coumaric acid + adriamycin group was given p-coumaric acid five days before adriamycin administration. Twenty-four hours after the last administration, blood samples were collected for biochemical analysis, and liver tissues were removed for histopathological and immunohistochemistrical studies. Moreover, the levels of tissue lipid peroxidation and enzyme activities of glutathione peroxidase, superoxide dismutase, and catalase in liver tissue were measured. Results: Treatment with p-coumaric acid protected the liver from the toxicity of adriamycin by attenuating the increase in alkaline phosphatase, alanine transaminase, aspartate transaminase, total bilirubin, total cholesterol, triglyceride, and low-density lipoprotein cholesterol and lessening the decrease in high-density lipoprotein cholesterol and albumin. p-Coumaric acid also raised the levels of glutathione peroxidase, superoxide dismutase, and catalase, as well as decreased lipid peroxidation in liver tissue and hepatic IL-1β expression. Additionally, histopathological study confirmed the protective effect of p-coumaric acid against liver damage. Conclusions: p-Coumaric acid can alleviate adriamycin-induced hepatotoxicity.

P-Coumaric acid alleviates adriamycin-induced hepatotoxicity in rats

Objective: To evaluate the effect of p-coumaric acid against adriamycin-induced hepatotoxicity in rats. Methods: The rats were divided into 4 groups. The control group received solvent; the p-coumaric acid group was treated with 100 mg/kg of p-coumaric acid orally for five consecutive days; the adriamycin group was administered with a single dose of adriamycin (15 mg/kg, i.p.), and the p-coumaric acid + adriamycin group was given p-coumaric acid five days before adriamycin administration. Twenty-four hours after the last administration, blood samples were collected for biochemical analysis, and liver tissues were removed for histopathological and immunohistochemistrical studies. Moreover, the levels of tissue lipid peroxidation and enzyme activities of glutathione peroxidase, superoxide dismutase, and catalase in liver tissue were measured. Results: Treatment with p-coumaric acid protected the liver from the toxicity of adriamycin by attenuating the increase in alkaline phosphatase, alanine transaminase, aspartate transaminase, total bilirubin, total cholesterol, triglyceride, and low-density lipoprotein cholesterol and lessening the decrease in high-density lipoprotein cholesterol and albumin. p-Coumaric acid also raised the levels of glutathione peroxidase, superoxide dismutase, and catalase, as well as decreased lipid peroxidation in liver tissue and hepatic IL- 1β expression. Additionally, histopathological study confirmed the protective effect of p-coumaric acid against liver damage. Conclusions: P-Coumaric acid can alleviate adriamycin-induced hepatotoxicity.

Directed evolution of tyrosine ammonia-lyase to improve the production of p-coumaric acid in Escherichia coli / 生物工程学报

p-coumaric acid is an important natural phenolic compound with a variety of pharmacological activities, and also a precursor for the biosynthesis of many natural compounds. It is widely used in foods, cosmetics and medicines. Compared with the chemical synthesis and plant extraction, microbial production of p-coumaric acid has many advantages, such as energy saving and emission reduction. However, the yield of p-coumaric acid by microbial synthesis is too low to meet the requirements of large-scale industrial production. Here, to further improve p-coumaric acid production, the directed evolution of tyrosine ammonia lyase (TAL) encoded by Rhodotorula glutinis tal gene was conducted, and a high-throughput screening method was established to screen the mutant library for improve the property of TAL. A mutant with a doubled TAL catalytic activity was screened from about 10,000 colonies of the mutant library. There were three mutational amino acid sites in this TAL, namely S9Y, A11N, and E518A. It was further verified by a single point saturation mutation. When S9 was mutated to Y, I or N, or A11 was mutated to N, T or Y, the catalytic activity of TAL increased by more than 1-fold. Through combinatorial mutation of three types of mutations at the S9 and A11, the TAL catalytic activity of S9Y/A11N or S9N/A11Y mutants were significantly higher than that of other mutants. Then, the plasmid containing S9N/A11Y mutant was transformed into CP032, a tyrosine-producing E. coli strain. The engineered strain produced 394.2 mg/L p-coumaric acid, which is 2.2-fold higher than that of the control strain, via shake flask fermentation at 48 h. This work provides a new insight for the biosynthesis study of p-coumaric acid.

Metabolic engineering strategies for caffeic acid production in Escherichia coli

Caffeic acid (CA; 3,4-dihydroxycinnamic acid) is an aromatic compound obtained by the phenylpropanoid pathway. This natural product has antioxidant, antitumor, antiviral, and anti-inflammatory activities. It is also a precursor of CA phenethyl ester (CAPE), a compound with potential as an antidiabetic and liver-protective agent. CA can be found at low concentrations in plant tissues, and hence, its purification is difficult and expensive. Knowledge regarding the pathways, enzymes, and genes involved in CA biosynthesis has paved the way for enabling the design and construction of microbial strains with the capacity of synthesizing this metabolite. In this review, metabolic engineering strategies for the generation of Escherichia coli strains for the biotechnological production of CA are presented and discussed.

P-Coumaric acid ameliorates ethanol-induced kidney injury by inhibiting inflammatory cytokine production and NF- and kappa;B signaling in rats

To examine the effects of p-coumaric acid on ethanol-induced kidney injury in Swiss Wistar rats. Methods: Ethanol (25% v/v) was used to induce nephrotoxicity in rats. p-Coumaric acid was orally administered at 50, 100, or 200 mg/kg body weight. The levels of oxidative parameters were determined; pro-inflammatory biomarkers were analyzed by Western blotting and apoptotic protein was analyzed by immunohistochemistry. Results: Ethanol treated rats showed decreased levels of antioxidants and aberrant production of pro-inflammatory cytokines (IL-6, IL1P, TNF-), NF- and kappa;B activation and imbalance of pro- and anti-apoptotic proteins (Bcl-2, Bax, caspase 3). Meanwhile, jp-coumaric acid restored antioxidant levels and decreased the levels of inflammatory cytokines, NF- and kappa;B, and pro- apoptotic proteins and increased Bcl-2 expression. Conclusions: jp-Coumaric acid ameliorates ethanol-induced kidney injury by restoring antioxidant production and suppressing cellular apoptosis and inhibiting NF- and kappa;B expression. jp-Coumaric acid should be further investigated as a promising candidate for ethanol-induced kidney toxicity.

Simultaneous determination of four effective components in Taraxaci Herba by UPLC-MS/MS / 国际药学研究杂志

Objective: To establish an LCMS/MS method for simultaneous determination of pcoumaric acid, caffeic acid,luteolin and luteoloside in Taraxaci Herba. Methods: The LC analysis was performed on a UPLC-MS/MS instrument with a Dikma Endeavorsil UPLC C18 column(100mm×2.1 mm,1.8 μm),the mobile phase was 0.1% formic acid solution-methanol in a gradient elution,and the flow rate was 0.2 ml/min;the column temperature was set at 35℃; the injection volume was 2 μl. For the MS/MS analysis,the ESI source was operated in the positive-ion mode and the ions were monitored in multiple reaction monitoring(MRM)mode,the temperature of drying gas was 370℃,the sheath gas pressure was set at 10 Bar,the aux gas pressure was maintained at 30 Bar,and the spray voltage was controlled at 4000 V. Results Calibration curves showed good linearity for all 4 compounds in the concentration range of 0.1-10 μg/ml,and the correlation coefficient®was 0.9991,0.9996,0.9994 and 0.9999 for p-coumaric acid,caffeic acid,luteolin,and luteoloside,respectively. Conclusion: The present method could be used for the simultaneous determination of the above mentioned 4 components in Taraxaci Herba.

p-Coumaric acid ameliorates ethanol-induced kidney injury by inhibiting inflammatory cytokine production and NF-κB signaling in rats

Objective: To examine the effects of p-coumaric acid on ethanol-induced kidney injury in Swiss Wistar rats. Methods: Ethanol (25％ v/v) was used to induce nephrotoxicity in rats. p-Coumaric acid was orally administered at 50, 100, or 200 mg/kg body weight. The levels of oxidative parameters were determined; pro-inflammatory biomarkers were analyzed by Western blotting and apoptotic protein was analyzed by immunohistochemistry. Results: Ethanol treated rats showed decreased levels of antioxidants and aberrant production of pro-inflammatory cytokines (IL-6, IL1β, TNF-α), NF-κB activation and imbalance of pro-and anti-apoptotic proteins (Bcl-2, Bax, caspase 3). Meanwhile, p-coumaric acid restored antioxidant levels and decreased the levels of inflammatory cytokines, NF-κB, and pro-apoptotic proteins and increased Bcl-2 expression. Conclusions: p-Coumaric acid ameliorates ethanol-induced kidney injury by restoring antioxidant production and suppressing cellular apoptosis and inhibiting NF-κB expression. p-Coumaric acid should be further investigated as a promising candidate for ethanol-induced kidney toxicity.

Effect of Sasa quelpaertensis Nakai Extracts and its Constituent p-coumaric Acid on the Apoptosis of Human Cancer Cell Lines

Sasa quelpaertensis Nakai leaves contain a mixture of polysaccharides, amino acids, and polyphenols, which mediate various biological activities. For efficient utilization of its leaf, we reported the preparation procedure for phytochemical-rich extract (PRE) using the leaf residue, which was by-product of hot water extraction. This study was undertaken to evaluate the effects of PRE and its major constituent, p-coumaric acid,on the growth of several human cancer cell lines (MKN-74, MKN-45, SNU-1, SNU-16, and HL-60). The ethyl acetate fraction of PRE and p-coumaric acid significantly inhibited the proliferation of MKN-74 and HL-60 cells, respectively, and induced cell apoptosis, down-regulated Bcl-2 and poly (ADP-ribose) polymerase levels, and up-regulated those of Bax and caspase-3. These results show the potential utility of S. quelpaertensis Nakai leaves in cancer prevention.

Antidepressant-like Effects of p-Coumaric Acid on LPS-induced Depressive and Inflammatory Changes in Rats

Depression causes mental and physical changes which affect quality of life. It is estimated to become the second most prevalent disease, but despite its commonness, the pathophysiology of depression remains unclear and medicine is not sufficiently protective. p-Coumaric acid (p-CA) is a dietary phenolic acid which has been proven to have antifungal, anti-HIV, anti-melanogenic, antioxidant and anti-inflammatory effects. Considering these effects, we investigated whether p-CA can prevent depressive symptoms by reducing inflammatory cytokines in animals injected with lipopolysaccharide (LPS). Changes in despair-related behaviors, inflammatory cytokines, neurotrophic factors and synaptic activity were measured. In these animals, p-CA improved despair-related behavioral symptoms induced by LPS in the forced swim test (FST), tail suspension test (TST) and sucrose splash test (SST). p-CA also prevented the increase of inflammatory cytokines in the hippocampus such as cycloxigenase-2 and tumor necrosis factor-α due to LPS. Similarly, it prevented the reduction of brain-derived neurotrophic factor (BDNF) by LPS. Electrophysiologically, p-CA blocked the reduction of long-term depression in LPS-treated organotypic tissue slices. In conclusion, p-CA prevented LPS-induced depressive symptoms in animals, as determined by behavioral, biochemical and electrophysiological measures. These findings suggest the potential use of p-CA as a preventive and therapeutic medicine for depression.

Research progress on pharmacological effects of p-coumaric acid / 中草药

p-coumaric acid is mainly found in fruits, vegetables, grains, and fungi, and is also abundant in Chinese herbal medicines. The pharmacological effects of p-coumaric acid has anti-oxidant, anti-inflammatory, immunomodulatory, antitumor effects, antiplatelet aggregation, cardiovascular protection, prevention and improvement of diabetes, and neuroprotection, while the anti-oxidant activities of p-coumaric acid is the important basis of other pharmacological effects. In addition, p-coumaric acid has a certain inhibitory effect on bacteria, and also can inhibit melanin formation and delay skin aging. This paper reviews the main pharmacological effects of p-coumaric acid and provides reference for the development of both medicinal and food resources.

Chemical constituents from roots of Eupatorium chinense / 中草药

Objective To study chemical constituents of the roots of Eupatorium chinense. Methods The chemical constituents were separated and purified by the normal phase silica gel column chromatography, preparative thin-layer chromatography, and preparative HPLC. Their structures were determined by various spectral data. Results Nineteen compounds were isolated from the acetic ether extract of E. chinense and the structures were identified as euparin (1), 1-[2-(1-acetoxymethyl-vinyl)-6-hydroxy-benzofuran-5-yl]- ethanone (2), 6-hydroxy-3β-methoxytrematone (3), euparone (4), 8-methoxy-9-hydroxythymol (5), dehydroespeleton (6), 8-methoxy-9-hydroxythymol 3-O-angelate (7), 9-hydroxythymol (8), 4-hydroxy cinnamic acid methyl ester (9), p-hydroxy benzaldehyde (10), 8,9-dehydro-10-hydroxythymol (11), diisobutyl phthalate (12), dibutyl phthalate (13), p-coumaric acid (14), dihydrocoumarin (15), methylcaffeate (16), 2,5-dimethylphenol (17), 1H-indazole (18), and (Z)-3-(hydroxymethyl)-7-methylocta- 2,6-dienly1 acetate (19). Conclusion The chemical constituents are investigated and identified from the roots of E. chinense for the first time. Among them, compounds 2, 3, 5-11 are isolated from E. chinense for the first time, and compounds 6, 9, 12, 14-19 are isolated from the genus of Eupatorium for the first time. Compounds 1-4 are benzofurans, which are the characteristic constituents in Eupatorium genus.

Effects of p-coumaric acid on preventing acute hypoxia-induced pulmonary edema mice / 中国药学杂志

OBJECTIVE: To study the preventing effects of p-coumaric acid(p-CA) on acute hypoxia-induced pulmonary edema by mice experiments. METHODS: Acute-hypoxia model was established using a normobaric hypoxia chamber in vivo. Salidroside was set as a positive control drug. And the test period was 7 d using a method of intragastric administration. The measurements including pulmonary water content, HE staining, inflammatory factors, anti-oxidative indexes and Na+, K+-ATPase were performed to determine the efficacies and mechanisms of p-CA on preventive against acute hypoxia-induced pulmonary edema. RESULTS: As compared with the normal group, pulmonary water contents increased significantly by 3.56% in the mice treated with acute hypoxia (9.5% O2) for 6 h (control group) (P<0.01), and administration with p-CA (25, 100 mg•kg-1•d-1) for 7 d could significantly reduce this index (P<0.05), which was as effective as the positive group. The action mechanisms of p-CA could be due to its abilities of improving the activity of Na+, K+-ATPase, enhancing antioxidant capacity (SOD↑, CAT↑ and MDA↓) and inhibiting inflammatory factors (IL-1β and IL-6). CONCLUSION: p-CA has greater preventive effects on acute hypoxia-induced pulmonary edema in mice.

Chemical constituents from aerial parts of Leonurus macranthus / 中草药

Objective: To investigate the chemical constituents from the aerial parts of Leonurus macranthus. Methods: The chemical constituents were separated and purified by silica gel, Sephadex LH-20, ODS column chromatographies, and semi-preparative HPLC. Their structures were determined by physicochemical properties and spectroscopic data. Results: Nineteen compounds were isolated from the CH2Cl2 layer of 70% aqueous acetone extract in the aerial parts of L. macranthus, and identified as (+)-syringaresinol (1), (+)-1-hydroxysyringaresinol (2), rayalinol (3), erythro-guaiacylglycerol-β-O-4'-coniferyl ether (4), (7R, 7'R, 7″S, 8S, 8'S, 8″S)-3', 4″- dihydroxy-3, 5, 4', 5″-tetramethoxy-7, 9': 7', 9-diepoxy-4, 8″-oxy-8, 8'-sesquineo-lignan-7″, 9″-diol (5), (7R, 7'R, 7″S, 8S, 8'S, 8″S)-4', 5″- dihydroxy-3, 5, 3', 4″-tetramethoxy-7, 9': 7', 9-diepoxy-4, 8″-oxy-8, 8'-sesquineo-lignan-7″, 9″-diol (6), genkwanin (7), 3'-hydroxy- genkwanin (8), eriodictyol (9), isoscopoletin (10), p-coumaric acid (11), caffeic acid methyl ester (12), trans-ferulic acid (13), syringic aldehyde (14), vanillic acid (15), oct-1-en-3-yl β-glucopyranoside (16), 5-hydroxy-2-pyrrolidone (17), pterolactam (18), and nicotinamide (19), respectively. Conclusion: Compounds 1-6 and 9-19 are isolated from the plants of genus Leonurus Linn. for the first time, and compounds 7 and 8 are found from L. macranthus for the first time.

Simultaneous Determination of P-coumaric Acid and Ferulic Acid in Different Compatibility Proportions of the Couple of Sparganium stoloni and Curcuma aeruginosa by HPLC / 中国药房

OBJECTIVE:To establish a method for simultaneous determination of P-coumaric acid and ferulic acid in the couple of Sparganium stoloni and Curcuma aeruginosa,and to investigate rational proportion of the couple.METHODS:HPLC method was adopted.The determination was performed on Agilent 20RBAX XDB-C18 column with mobile phase consisted of acetonitrile-0.1％ phosphoric acid (gradient elution) at the flow rate of 1.0 mL/min.The detection wavelength was set at 266 nm,and the column temperature was 30 ℃.The sample size was 10 μL RESULTS:The linear ranges of P-coumaric acid and ferulic acid were 4.218 6-21.093 μg/mL(r=0.999 8),1.836 0-9.180 μg/mL(r=0.999 9),respectively.RSDs of precision,stability and reproducibility tests were all lower than 2.0％.The recoveries were 98.72％-100.30％ (RSD=0.18％,n=9),99.11％-100.45％ (RSD=0.46 ％,n =9).Among 4 compatibility proportions of the couple of S.stoloni and C.aeruginosa (1 ∶ 1,2 ∶ 1,1 ∶ 2,1 ∶ 0,m/m),the dis-solution rate of P-coumaric acid and ferulic acid were the highest when the ratio of S.stoloni-C.aeruginosa (m/m)was 2 ∶ 1.CON-CLUSIONS:The method is simple,accurate,stable and reproducible,and can be used for content determination of P-coumaric acid and ferulic acid in the couple of S.stoloni and C.aeruginosa.when the proportion of the couple of S.stoloni and C.aeruginosa is 2∶1 and the contents of P-coumaric and ferulic acid are the highest,the contents of main etfective components are the highest.