ABSTRACT
Pyrrosia petiolosa, Pyrrosia lingua and Pyrrosia sheareri are recorded as original plants of Pyrrosiae Folium (PF) and commonly used as Chinese herbal medicines. Due to the similar morphological features of PF and its adulterants, common DNA barcodes cannot accurately distinguish PF species. Knowledge of the chloroplast (cp) genome is widely used in species identification, molecular marker and phylogenetic analyses. Herein, we determined the complete cp genomes of three original species of PF via high-throughput sequencing technologies. The three cp genomes exhibited a typical quadripartite structure with sizes ranging from 158 165 to 163 026 bp. The cp genomes of P. petiolosa and P. lingua encoded 130 genes, whilst that of P. sheareri encoded 131 genes. The complete cp genomes were compared, and five highly divergent regions of petA-psbJ, matK-rps16, ndhC-trnM, psbM-petN and psaC-ndhE were screened as potential DNA barcodes for identification of Pyrrosia genus species. The phylogenetic tree we obtained indicated that P. petiolosa and P. lingua are clustered in a single clade and, thus, share a close relationship. This study provides invaluable information for further studies on the species identification, taxonomy and phylogeny of Pyrrosia genus species.
ABSTRACT
To screen the active fractions with lithagogue effects of Pyrrosia lingua from Guizhou province and preliminarily investigate its mechanism. The rats were fed with 1% ethylene glycol and 2% ammonium chloride to establish the nephrolithiasis models, which were used to evaluate thelithagogue effects of different polar fractions of P. lingua from Guizhou province. The level of urinary calcium and oxalic acid in urine, renal calcium, oxalic acid, superoxide dismutase (SOD), catalase(CAT) and malondialdehyde (MDA) in renal tissues,as well as crystalline deposit and lithogenesis in renal tissues and the levels of creatinine(Cr) and blood urea nitrogen (BUN) in the serum were detected. The effective compounds were inferred from the analysis of active fractions extract based on LC-MS technology. Petroleum ether fraction and dichloromethane fraction of P. lingua from Guizhou province can reduce renal oxalic acid and renal calcium concentration, increase urinary oxalic acid and urine calcium, with significant inhibitory effect on the formation of renal calculus in rats, significantly increase SOD and CAT activities in renal tissues, and significantly reduce MDA levels. LC-MS analysis showed that the caffeine, citric acid and tartaric acid among the compounds from petroleum ether fraction and dichloromethane fraction had lithagogue effects. Both the petroleum ether fraction and dichloromethane fraction of P. lingua from Guizhou province showed good effect on prevention and treatment of calculus in middle dose groups, and the mechanism may be associated with antioxidation, reducing calcium oxalate crystal deposition, and promoting calcium oxalatecrystal release, in addition, caffeine, citric acid and tartaric acid had lithagogue effects.
ABSTRACT
OBJECTIVE:To establish a method for the contents determination of 4 amino acids in Pyrrosia lingua. METHODS:The samples were analyzed by RP-HPLC. It was performed on a column of Welchrom C18 with the mobile phase A of 0.1 mol/L So-dium acetate buffer(pH6.5)- acetonitrile(93∶7,V/V)and mobile phase B of acetonitrile-water(4∶1,V/V)(gradient elution)at the flow rate of 1.0 ml/min,detection wavelength was 254 nm,column temperature was 30 ℃ and the sample size was 5 μl. RE-SULTS:The linear range was 4.92-49.2 μg/ml for glutamate(r=0.999 1),3.168-31.68 μg/ml for glycine(r=0.999 9),2.6-26 μg/ml for alanine(r=0.999 4)and 2.88-28.8 μg/ml for leucine(r=0.999 9);RSDs of precision,stability and reproducibility tests were no more than 2.2%;average recoveries were 98.4%(RSD=2.2%,n=9),99.6%(RSD=2.5%,n=9),98.3%(RSD=2.2%,n=9)and 100.1%(RSD=2.3%,n=9),respectively. CONCLUSIONS:The method is simple,accurate and reproducible,and can be used for the contents determination of 4 amino acids in P. lingua.
ABSTRACT
Objective To establish an HPLC method for simultaneous separation and determination of chlorogenic acid, quercetin and kaempferol inPyrrosia lingua (Thumb.) Farwell. Methods The separation was performed on Phenomenex C18 column (250 mm×4.6 mm, 5μm) with the mobile phase of methanol-acetic acid (pH=3.0) solution and gradient elution. Flow rate was 1.0 mL/min;the UV detection wavelength was 254 nm;column temperature was 35℃.Results The calibration curves for chlorogenic acid, quercetin, and kaempferol were in good linearity in the range of 0.000 24-3.00μg (r=0.999 9), 0.000 16-2.00μg (r=0.999 9), and 0.000 16-2.00μg (r=0.999 9), respectively. The limits of detection (S/N=3) were 3.29, 0.43 and 0.33 ng/mL, respectively. The average recovery rates were 97.73%, 98.07% and 96.92%, respectively. ConclusionThe method is simple, precise and sensitive. It provides scientific proof for separation and determination of chlorogenic acid, quercetin and kaempferol inPyrrosia lingua (Thumb.) Farwell.