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Objective: To study the constituents from the roots of Paeonia lactiflora. Methods: The chemical constituents were isolated by various chromatographic techniques and their structures were elucidated by physicochemical properties and NMR data. Results: Eleven compounds were obtained and characterized as (4S)-perillic acid 6-O-α-L-arabinopyranosyl-(1'→6'')- β-D-glucopyranosyl (1), 4,9-dihydroxy-8,10-dehydrothymol-1-O-β-D-glucoside (2), emodin-8-O-β-D-glucoside (3), resveratrol (4), β-D-glucopyranosyl benzoate (5), ilexperphenoside A (6), catechol (7), methyl gallate (8), ethyl gallate (9), 3-methoxygallic acid (10), 1,2,3,4,6-penta-O-galloyl-β-D-glucopyranoside (11). Conclusion: Compound 1 is identified as a new compound, named perillic acid glycoside, compounds 2 and 5 are identified from the genus Paeonia for the first time.
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OBJECTIVE: To establish an HPLC method for simultaneous determination of 15 monoterpene glycosides in Radix Paeoniae Alba and seed cake of peony. METHODS: The separation was performed on an Agilent Zorbax SB C18-Aq column, using acetonitrile (A) and potassium dihydrogen phosphate solution (B) (pH 2.8) as the mobile phase by gradient elution. The elution program was as follows: 0 min(A, 9%)→8 min(A, 9%)→10 min(A, 15%)→25 min(A, 20%)→42 min(A, 35%)→50 min(A, 35%). The flow rate was 1.0 mL•min-1. The detection wavelength was maintained at 260 nm. The column temperature was set at 30 ℃. RESULTS: The linear range was 5.1-81.6 μg•mL-1 for pyrindylpaeoniflorin, 12.95-207.2 μg•mL-1 for mudanpioside F, 6.2-99.2 μg•mL-1 for oxyalbiflorin, 12.2-195.2 μg•mL-1 for oxypaeoniflorin, 8.0-128.0 μg•mL-1 for 10-hydroxypaeoniflorin, 5.5-88.0 μg•mL-1 for albiflorin, 6.0-96.0 μg•mL-1 for paeoniflorin, 6.0-96.0 μg•mL-1 for oxypaeonidanin, 5.6-89.6 μg•mL-1 for 4-O-methyl-oxypaeoniflorin, 4.7-75.2 μg•mL-1 for galloylpaeoniflorin, 5.4-86.4 μg•mL-1 for 4-O-methyl-paeoniflorin, 5.0-80.0 μg•mL-1 foralbiflorin R1, 5.7-91.2 μg•mL-1 for paeonidanin, 5.4-86.4 μg•mL-1 for benzoyloxypaeoniflorin and 6.7-107.2 μg•mL-1 for benzoylpaeoniflorin. The average recoveries of the 15 monoterpene glycosides, were between 96.8%-105.6% and the RSD values were 1.05%-3.41%. CONCLUSION: The method is simple, accurate and can be used for the quality control of peony.
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Objective This study focused on the secondary metabolites of endophytic fungus Alternaria alternate in Paeonia lactiflora. Methods Compounds were isolated from the EtOAc extract by chromatography technology and their structures were elucidated on the basis of comprehensive spectroscopic analysis. Results A total of 15 compounds were isolated and their structures were identified as methyl 4-acetamido-3-hydroxybenzoate (1), (E)-methyl5-hydroxy-3-methylpent-2-eno (2), isobenzofuranone A (3), 4-hydroxyacetophenone (4), 6-hydroxy-isosclerone (5), talarojlavone (6), 7-hydroxy-2-hydroxymethyl-5-methyl-4H-chromen-4-one (7), alternarienonic acid (8), 7-hydroxy-2,5-dimethyl-4H-1-benzopyran-4-one (9), 5-hydroxy-epialtenuene (10), alternariol (11), methyl 3-hydroxybenzoate (12), stemphyperylenol (13), altenusin (14), and altenuene (15). Conclusion Compounds 1, 3, 5-10, 12, and 13 are isolated from Alternaria alternatefor the first time.
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Objective To develop a method for the measurement of the content of paeoniflorin and albiflorin in red peony root (the roots of Paeonia lactiflora, RPR) by near-infrared spectroscopy (NIR) for quality rapid assessment. Methods A total of 65 RPR samples were collected from Sichuan, Zhejiang and Anhui of China, and then quantified the content of paeoniflorin and albiflorin by NIR and ultra-high performance liquid chromatography (UPLC). Fifty-three samples were chosen as the calibration set, while the remaining 12 samples were assigned to external validation set. The calibration model of paeoniflorin and albiflorin was further developed by modified partial least squares (MPLS) using UPLC data as reference based on optimized spectral regions, pre-treament of spetrum and the number of principal divisors. The content of paeoniflorin and albiflorin in unknown samples (validation model) was predicted according to the validation set of NIR. Results The standard errors of prediction (SEP) for the contents between the predicted value of NIR method and the estimated value of UPLC method were 1.437 2% for paeoniflorin and 0.784 3% for albiflorin, and their correlation coefficient (R) were 0.990 2 for paeoniflorin) and 0.994 4 for albiflorin in 53 calibration set samples. Moreover, the SEP and R of that in 12 validation set samples were 0.714 5%, 0.988 0% for paeoniflorin, and 0.632 4, 0.994 6 for albiflorin, respectively. The sum of paeoniflorin and albiflorin in RPR cultivated in Sichuan, Anhui, and Zhejiang province in China were 5.49%, 5.33%, and 4.81%, respectively. Conclusion NIR can quantify the amounts of paeoniflorin and albiflorin quickly and simultaneously in RPR for quality rapid assessment. The quality of RPR cultivated in Sichuan, Anhui, and Zhejiang is similar based on the sum of these two bioactive compounds without significant differences.
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Objective: To improve the dissolution of Salviae Miltiorrhizae Radix et Rhizoma (SMRR) and Paeoniae Radix Alba (PRA) in Tongmai Dasheng Tablets (TDT), and to construct a good powder dispersion. Methods: SMRR, PRA, and Placenta Hominis (PH) were crushed by common method and ultramicro method using calgon as a dispersant to prepare 19 powder dispersion samples. The particle size distribution, infrared spectrum, dispersion, specific surface area, and porosity of the powder dispersions were investigated. The contents of salvianolic acid B and paeoniflorin were measured by HPLC, and the dissolution of salvianolic acid B and paeoniflorin in different powder dispersions was evaluated. Results: Infrared spectrum results showed that superfine pulverization, mixed comminution or adding the auxiliary pulverizing cannot produce new substances in medicinal powder. The dissolution rate of salvianolic acid B and paeoniflorin in the mixed ultramicro powder dispersions of SMRR and PRA was the highest, salvianolic acid B 83.57%, RSD 2.03%, paeoniflorin 80.75%, RSD 0.61%. Conclusion: The dissolution rate is affected not only by the specific surface area of the powder but also by the dispersity of the powder. In order to improve the dissolution rate of active ingredients in powder, it is important to increase the specific surface area of the powder and improve the dispersity of the powder. In the mixed powder of SMRR, PRA, and PH, the dissolution rate of salvianolic acid B and paeoniflorin was lower than that of the mixed powder of SMRR and PRA. Hence, PH was smashed alone, and then mixed with the other two kinds of medicinal powder.
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Objective: To establish an ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method for the simultaneous determination of 23 plant growth regulator (PGR) residues in Chinese materia medica (CMM). Methods: Samples were extracted with acetontrile, and then determined by UPLC-MS/MS directly without further clean up. The matrix-matched external standard method was used for quantitative analysis. Results: The calibration curves showed good linearity in each range with correlation coefficients greater than 0.99. The limits of detection (LODs) were 0.01-20.80 ng/mL for the 23 PGRs spiked in Codonopsis pilosula and Angelica dahurica. The recoveries of the 23 PGRs spiked in C. pilosula and A. dahurica at three levels of 0.01, 0.04, and 0.1 mg/kg were in the range of 71.0%-101.4%, the relative standard deviation (RSDs) were 0.8%-15.2%. Commonly used CMM including eight species (63 batches) was analyzed by this method. Conclusion: The method proved to be simple, rapid, sensitive, accurate, and suitable for the simultaneous determination of 23 PGR residues in CMM
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Objective: To clone the full-length cDNA sequence which encodes one of the key enzymes of terpene biosynthesis, farnesyl pyrophosphate synthase (FPPS) and analyze the bioinformation. Methods: Based on the transcriptome data of Paeonia lactiflora (Pl), a pair of specific PCR primers were designed. The objective PCR band of PlFPPS was successfully amplified using RT-PCR technique and then it was cloned to pMD18-T vector. After clone and plasmid PCR characterization, the recombinants were sequenced and then a series of bioinformatic analysis was carried out including sequence homology search, construction of molecular phylogenetic tree, domain search, and 3D structure prediction, etc. Results: Sequencing results showed that the full-length PlFPPS was 1 315 bp, which contained 60 bp 5'-UTR, 1 050 bp CDS, and 205 bp 3'-UTR. It encoded 349 amino acids and the accession number of GenBank was KP708571.Blastn and Blastp analysis revealed that PlFPPS (KP708571) and its encoding protein (AKJ26301) had high homology with FPPS genes and proteins from several plants at the nucleotide and amino acid levels. Phylogenetic tree analysis indicated that the genetic relationship of PlFPPS with FPPS from other plants was relatively far. It predicted that the molecular weight and isoelectric point of PlFPPS were 40200 and 5.33, which was a hydrophilic and stable protein, located in cytoplasm without transmembrane domain and signal peptide. It contained seven conservative domains like substrate binding pocket, substrate-Mg2+ binding site, catalytic site, aspartate-rich region 1, and aspartate-rich region 2.The proportion of α-helix in its 3D structure was high and the α-helix was linked by β-loop. The central cavity was composed of 10 core α-helixes. Conclusion: The full-length cDNA sequence of FPPS is cloned firstly from P. lactiflora and the bioinformatic analysis is then carried out.
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Objective: To construct the prokaryotic expression vector of Actin gene and to establish the high-level expression of Actin of Paeonia lactiflora in Escherichia coli. Methods: Based on cDNA sequence of Actin of P. lactiflora and the polyclonal sites on the prokaryotic expression vector pET-32a (+), a pair of PCR primers whose 5' end was inserted with BamH I and Hind III, respectively were designed. Subcloning of Actin was carried out using RT-PCR technique. The recombinant plasmid pMD18-T-PlActin and the prokaryotic expression vector pET-32a (+) was digested by BamH I and Hind III and the objective gene and the empty vector were purified. After ligation and transformation, the recombinant pET-32a (+)-PlActin, which was characterized by colony PCR, plasmid PCR, and double enzyme digestion, was transformed into BL21 (DE3), and the engineering expression strain was obtained. The expression of recombinant Actin protein in E. coli was induced at different concentration of inducer IPTG, different bacteria density, and different expression time. After SDS-PAGE and Coomassie brilliant blue R250 staining, the dry gel was prepared and the expression level of recombinant Actin protein was analyzed. Results: Subcloning sequencing result showed that the sequence of PlActin was exactly same with the objective sequence and the 5' and 3' ends were successfully inserted with BamH I and Hind III sites. The prokaryotic expression vector of Actin gene pET-32a (+)-PlActin was constructed successfully. The best concentration of IPTG was 0.1 mmol/L and the bacteria density A600 was 0.4 to 0.6. The optimal expression time was 4 h. Conclusion: The prokaryotic expression vector of Actin gene pET-32a (+)-PlActin is constructed and the high-level expression of Actin of P. lactiflora in E. coli is established.
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Objective: Genomic gene encoding Actin in Paeonia lactiflora was cloned in order to clarify the gene organization and its expression levels in different tissues in herbaceous peony. Methods: Based on cDNA sequences of Actin genes isolated from P. lactiflora reported by our laboratory, one pair of PCR primers was designed. PCR products of Actin genomic gene were successfully amplified with total genomic DNA extracted from herbecous peony cv. "Taohuafeixue" as template by high-fidelity DNA polymerase KOD-Plus, and then they were cloned to pMD18-T vector and be sequenced. The exons and introns of Actin gene were predicted with bioinformatic softwares. The homology was analyzed by Blastn at the nucleotide level and the molecular phylogenetic tree was constructed with MEGA5.0 software. Based on sequencing results, one pair of PCR primers was designed, and the expression levels of Actin gene in the roots, stems, flowers, and leaves in herbaceous peony were semi-quantified. Results: The sequencing results showed that Actin gene of herbaceous peony was 1405 bp length, which contained four exons and three introns. All the splicing sites in three introns were conservative at 5' donor with GU and 3' receptor with AG. It encoded 377 amino acids, and GenBank accession number was KF363830. A pair of PCR primers was designed and one of them was spaned the first intron, which could effectively prevent the false postive by genomic DNA contamination. The semi-quantitative RT-PCR analysis showed that the expression levels of Actin gene in the roots, stems, leaves, and flowers of herbaceous peony were almost constant. Conclusion: It is the first report on cloning and gene organization clarification of genomic gene encoding Actin in P. lactiflora. The semi-quantitative analyses indicate that Actin gene can be used as the internal standard gene for the expression analysis of the functional genes in herbaceous peony.
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Paeonia lactiflora Pall. has a long history of utilization, and is widely used in pharmaceutical, food and cosmetic industry. In this paper, the invention patents of traditional medicine P. lactiflora before 2014 were retrieved, and totally 18 192 patent families were obtained. And we formed the patent analysis report of medicine P. lactiflora based on a multi-angle analysis. Results show that, as to the patent number of medicine P. lactiflora, China is much more than any other country, and the applications mainly came from pharmaceutical enterprises. But the technologi-cal quality of patents and international protection ratio are low in our country. We need to strengthen in treatment of cardiovascular and liver disease. The patents mainly focused on the use of tonic health, which is compatible with Traditional Chinese Medicine. On the product development, dosage forms need to be enhanced. As to the compre-hensive utilization of resources, the flower and seed of P. lactiflora have relatively larger research space and value. This work will be helpful for researchers in deeply understanding the research achievements of medicine P. lactiflora, and provides the reference data for the future research.
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Background: We previously evaluated the biochemical changes induced by the local product TCM for diabetes (TCM-D™) on blood glucose levels and other biochemical changes in normal mice fed orally with the recommended human dose (30 ml/kg daily) and ten times this dose for eight weeks. TCM-D™ is an aqueous extract of the roots of Trichosanthes kirilowii Maxim, Paeonia lactiflora Pall, Glycyrrhiza uranlensis Fisch. and Panax ginseng Meyer (red) combined at the dry weight proportions of 36%, 28%, 18% and 18% respectively. The study showed that at these dosages the blood glucose levels as well as the body weights in treated mice were significantly reduced when compared with pretreatment values and control animals. The present study evaluated the effect of the extract in a mouse model of Type 1 diabetes mellitus. Methods: TCM-D™ extract was prepared as a 10x concentrate and given orally at 0.3 ml/100 g and 1.5 ml/100 g to mice which were experimentally induced diabetic with intraperitoneal injections of streptozotocin (5 mg/100g) in sodium citrate (pH 4.5). Control diabetic mice were dosed with extract diluent (distilled water). Results: At the doses studied the compound did not show any significant lowering of the glucose levels in a mouse model of Type 1 diabetes. There were significant increases in the alanine aminotransferase (ALT) and creatinine levels which were most likely due to the treatment with the compound. There were no significant changes in the aspartate aminotransferase (AST) and blood urea levels due to the treatment. Neither was there any significant effect on the weight of the treated animals due to the treatment. Conclusions: It is concluded that TCM-D™ did not have any significant blood glucose lowering effect on streptozotocin induced diabetic mice when fed orally at 1-5 times the recommended human dose. Further work is needed to determine if the extract has any significant effect in a mouse model with Type 2 diabetes mellitus.
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Objective: Cloning of Actin gene from Paeonia lactiflora and expression analysis of Actin gene with RT-PCR technique. Methods: Based on cDNA sequences of Actin genes isolated from the related species reported in GenBank, one pair of PCR primers was designed. PCR products of Actin gene were successfully amplified with RT-PCR technique using cDNA synthesized from the total RNA extracted from the roots of P. lactiflora 'Taohuafeixue' as template, and then they were cloned to pMD18-T vector with TA cloning method. The positive clones which were characterized with colony PCR, plasmid PCR, and enzyme digestion method were sequenced. Based on the sequencing results, one pair of PCR primers was designed, and the expression profile of Actin gene in the roots, stems, flowers, and leaves in P. lactiflora were semi-quantified with RT-PCR technique. Results: The sequencing results showed that the length of Actin gene of P. lactiflora was 1134 bp, encoding 377 amino acids, whose GenBank accession number was JX310002. Sequence analysis showed that Actin of P. lactiflora contained three kinds of characteristic signal sequences, whose homologous similarity in amino acid level with other plants was up to 99%. Based on homology modeling analysis, it revealed that there were four domains in its 3D structure. Bioinformatic software predicted that the molecular weight of Actin of herbaceous peony was 4.17 × 104, and the isoelectric point (pI) was 5.31. It was a hydrophilic and stable protein which was located in cytoplasm, without the transmembrane domain or signal peptide. Semi-quantitative RT-PCR analysis showed that the gene expression levels of Actin gene in the roots, stems, flowers, and leaves of P. lactiflora were almost constant. Conclusion: It is the first report on the cloning of Actin gene from P. lactiflora and the semi-quantitative analysis indicates that Actin gene can be used as the internal standard gene for the expression analysis of functional genes in P. lactiflora. This project plays a base for the effective application of Actin gene in P. lactiflora.
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Background: A number of Traditional Chinese Medicine (TCM) preparations are being used for the treatment of diabetes mellitus. Some components of these preparations have biochemical effects other than those of lowering blood glucose and indeed have been used for other medical indications in traditional practice. The primary objective of the study was to determine the effect of the oral mixture of Traditional Chinese Medicine for diabetes (TCM-D™ complex) on blood glucose level and the biochemical changes if any, on the liver (ALT, AST, gamma-GT, albumin, globulin) and renal (blood creatinine, urea) functions in normal mice. The oral mixture is an aqueous extract of four wellknown traditional Chinese medicinal herbs and consists of Trichosanthes kirilowii Maxim., Paeonia lactiflora Pall.,Glycyrrhiza uranlensis Fisch., and Panax ginseng (red) CA Meyer in the proportion of 36%, 28%, 18%, and 18% respectively of the dry weight. These herbs have been shown to have blood glucose lowering activity and have been used for other traditional medicinal purposes. The safety of the combination was evaluated in the present study. Methods: Experimental Balb/c mice were treated orally via gastric tube with the extract at daily doses equivalent to 1 and 10 times the recommended human dose for 8 weeks. Blood glucose and other biochemical profiles were monitored at pre-treatment and monthly posttreatment until killed. Results: When compared to pre-treatment levels, the blood glucose levels were significantly lower in treated animals compared to those in the control group. At the recommended TCM-D™ dose the levels in treated animals were significantly lower than that of control animals and at pre-treatment. When compared with pre-treatment, the glucose levels were lowest at Week 8 of treatment, the mean levels being 111.23%, 83.32% and 70.33% in control, and in animals given 1 x and 10 x the recommended TCM-D™ dosage respectively. The blood glucose lowering effect was also associated with a significant weight loss in treated animals. There were transient increases in AST and ALT levels but these reverted to normal at Week 8 of treatment. The levels of bilirubin, g-GT, albumin, creatinine and blood urea were also not significantly different at Week 8 from pre-treatment levels in all groups. Conclusion: Even at 10 times the dosage recommended for humans, TCM-D™ did not affect the liver and renal functions of treated animals. Treated and control animals remained healthy and normal throughout the period of observation.
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ObjectiveTo research the metabolism of components in the Radix of Paeonia lactiflora Pall. Methods(①) we established the HPLC fingerprint of water extract of Paeonia lactiflora Pall. and real-time monitored the chemical composition. (②) We established the HPLC fingerprint of rats' serum samples from hepatic portal vein, serum samples from aorta abdominalis and samples of intestinal absorption of Paeonia lactiflora Pall. (③) On this basis, using the established methods, I-IPLC fingerprint spectrum of serum samples,the sample of herb, the sample after intestinal metabolism, rats' serum samples from hepatic portal vein and rats'serum samples from aorta abdominalis were analyzed and compared in order to infer the metabolism of components in the Radix of Paeonia lactiflora Pall. Results24 compositions were detected, seven of which were metabolized by intestinal flora and could not be absorbed into blood; six of them could not be absorbed directedly into intestinal; eight new compounds were absorbed into blood after bowel metabolism while they were not detected in water extract in Paeonia lactiflora Pall. ConclusionWe could infer the metabolic processes of chemicals of Paeonia lactiflora Pall. in oral administration with this method.
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Objective To establish a HPLC method for determining the content of Paeonol in Paeonia lactiflora Pall.. Methods Ultrasound and reflux methods were used to extract Paeonol in Paeonia lactiflora Pall., and then the content of Paeonol was determined by HPLC. It was performed on Agilent column ZORBAX SB-C18 (Stable Bond Analytical 4.6 mm?250 mm, 5 ?m), mobile phase was methanol-water (45∶55), flow rate was 1 mL/min, column temperature was 25 ℃, and detection wavelength was 274 nm. Results The regression equation was Y=1.01?105X-9.84?102, r=0.999 9 (n=5). The standard curve was linear within the range of 0.104～0.52 ?g for Paeonol. Conclusion The method is simple and accurate with good reproducibility. It is suitable for determining the content of Paeonol in Paeonia lactiflora Pall. and control the quality control of Paeonia lactiflora Pall. in process.
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This study was conducted to investigate the antioxidative effects of Paeonia lactiflora (PL) seeds on antioxidative defense system and lipid peroxidation of liver in rats fed high-cholesterol diet. Sprague-Dawley male rats weighing 100+/-10g were randomly assigned into five experimental groups fed 0.5% cholesterol ; HC group which was not supplemented PL seeds extract, 0.1% methanol extract diet group (MP1 group), 0.2% methanol extract diet group (MP2 group), 0.05% ether-souble fraction diet group (EP1 group) and 0.1 % ether-souble fraction diet group (EP2 group). Experimental diets were fed ad libitum to the rats for 3 weeks. The activity of hepatic superoxide dismutase (SOD) was not significantly different among all the high cholesterol diet groups. The hepatic glutathione peroxidase (GSHpx) activity in MP2 group was increased to 27% compared to HC group. The activity of hepatic catalase (CAT) was not significantly different among the all high cholesterol diet groups. The hepatic glutathione S-transferase (GST) activity in the EP1 and EP2 groups were increased to 12% and 13%, respectively, as compared to HC group. The levels of hepatic TBARS in the MP1, MP2, EP1 and EP2 groups were reduced by 18%, 21%, 20% and 23%, respectively, as compared with HC group. The contents of lipofuscin in liver was not significantly different among all the experimental groups. The results indicated that PL seeds extract may be reduced oxidative damage by activating antioxidative defense system of hepatic in rats fed high-cholesterol diets.
Subject(s)
Animals , Humans , Male , Rats , Catalase , Cholesterol , Diet , Glutathione Peroxidase , Glutathione Transferase , Lipid Peroxidation , Lipofuscin , Liver , Methanol , Paeonia , Rats, Sprague-Dawley , Superoxide Dismutase , Thiobarbituric Acid Reactive SubstancesABSTRACT
Objective To investigate the dormancy reason of the seeds by studying on intrinsic inhibitor in the seed of Paeonia lactiflora.Methods Biological methods and embryo culture were carried out to study the activity of intrinsic inhibitor in the testa and endosperm of the P.lactiflora seeds.ResultsThe Activity of intrinsic inhibitor exists in the testa and endosperm,and both of the intrinsic inhibitors showed stronger inhibitory effect on the growth of younger roots of Brassica pekinensis than that on their seed germination.The Rf 0.5 section of the ether extract of endosperm showed the strongest inhibition on the growth of younger roots of B.pekinensis(relative rate: 36.36%).All embryos that inoculated on different culture medium could root,but the epicotyls of embryos that inoculated on culture medium without GA3 could not elongate,which showed that the embryo in vitro also exists the dormancy of epicotyl,or embryo with inhibitor.Conclusion Intrinsic inhibitor in the seed of P.lactiflora is the main factor that results in its dormancy.
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Total glucosides of paeony ( TGP ) , an extract of Paeonia Lacti-flora Pall were investigated for its effect on the synthesis of leukotriene B_4 ( LTB_4 ) by peritoneal macrophages ( PM?). LTB4, generated by 10~9~15 ? 10~10/L PM? with ionophore A_23187 , were measured with RP-HPLC. Suppression of releasing LTB_4 from PM? by TGP ( 100 mg/L ) was as much as the same dose of Flufenamic acid, but the action of TGP was slower. TGP significantly inhibited the synthesis of LTB_4 by PM? in a dose-dependent manner. The concentration of TGP required to obtain 50% inhibition ( IC_50 ) of formation of LTB_4 was 0.66mg/L. The data suggested antiinflam-matory and immunomodulatory action of TGP were related to its suppression of releasing LTB_4