ABSTRACT
OBJECTIVE：To establish the fingerprint of raw produc ts and different processed products of Pinellia pedatisecta ， to determine the contents of 5 kinds of nucleosides ，and to compare the differences of components between the raw products and processed products. METHODS ：P. pedatisecta raw products ，processed products by Processing Standard of Chinese Medicine in Henan Province （called“Standard processed product ”for short ）and processed products by new integrated processing technology in the production area （called“new integrated processed product ”for short ）were collected as investigation objects （12 batches of each）. The determination was performed on SymmetryShield RP 18 column with mobile phase consisted of acetonitrile （A）-0.1％ acetic acid aqueous water solution （B）（gradient elution ）at the flow rate of 0.8 mL/min，with the column temperature of 30 ℃， the detection wavelength of 270 nm，and the injection volume of 15 µL. HPLC fingerprints of 3 kinds of P. pedatisecta samples were established by using Similarity Evaluation System of TCM Chromatographic Fingerprints （2004 A version ） ，and the similarity of fingerprints was evaluated. The chromatographic peaks were identified by comparing with the reference chromatogram. Five nucleosides （adenine，hypoxanthine，uridine，xanthine，inosine）were quantitatively analyzed. SPSS 21.0 software was used for cluster analysis of 36 batches of samples. RESULTS ：The results of fingerprint and content determination met the relevant requirements. The similarity of 3 kinds of sample with their control fingerprint were all greater than 0.990. There were 22 common peaks in the raw products of P. pedatisecta ，and 16 common peaks were identified in the 2 kinds of processed products （the same 6 peaks disappeared from 2 kinds of processed products ）. Fivecomponents were identified in 3 kinds of samples ，such as adenine（peak 3），hypoxanthine（peak 7），uridine（peak 8）， 1064056472@qq.com xanthine（peak 9）and inosine （peak 11）. Results of content determination showed that total contents of 5 kinds of nucleosides in 2 kinds of proc essed products were all· decreased；the contents of them in descending order w as raw product ＞new i ntegrated processed products ＞Standard processed products. Results of cluster analysis showed that 36 batches of samples could be clustered into 2 categories，i.e. raw product was clustered into one category and 2 kinds of processed products into other one . CONCLUSIONS ：Established method is stable ， feasible and suitable for the quality evaluation of raw products and different processed products of P. pedatisecta . Fingerprints have changed significantly and the total content of 5 kinds of nucleosides in P. pedatisecta are all decreased after processing ，but that of new integrated processed products is slightly higher than that of Standard processed products .
ABSTRACT
The present study was designed to determine the chemical constituents of the stem tuber of Pinellia pedatisecta. The chemical constituents were isolated and purified by various chromatographic techniques, and their structures were elucidated on the basis of physicochemical properties and spectral data. Three new alkaloids (compounds 1, 2, and 3) were obtained and identified as 9-((5-methoxypyridin-2-yl)methyl)-9H-purin-6-amine (1), 4-(2-(2, 5-dioxopyrrolidin-1-yl)ethyl)phenyl acetate (2), and N-(9-((5-methoxypyridin-2-yl)methyl)-9H-purin-6-yl)acetamide (3). These compounds were evaluated for their cytotoxicity against human cervical cancer HeLa cells. Compounds 1 and 3 significantly inhibited the proliferation of HeLa cells with IC values being 3.02 ± 0.54 and 7.16 ± 0.62 μmol·L, respectively.
Subject(s)
Humans , Alkaloids , Chemistry , Pharmacology , Cell Proliferation , HeLa Cells , Pinellia , Chemistry , Plant Extracts , Chemistry , Pharmacology , Plant Stems , Chemistry , Plant Tubers , ChemistryABSTRACT
The present study was designed to determine the chemical constituents of the stem tuber of Pinellia pedatisecta. The chemical constituents were isolated and purified by various chromatographic techniques, and their structures were elucidated on the basis of physicochemical properties and spectral data. Three new alkaloids (compounds 1, 2, and 3) were obtained and identified as 9-((5-methoxypyridin-2-yl)methyl)-9H-purin-6-amine (1), 4-(2-(2, 5-dioxopyrrolidin-1-yl)ethyl)phenyl acetate (2), and N-(9-((5-methoxypyridin-2-yl)methyl)-9H-purin-6-yl)acetamide (3). These compounds were evaluated for their cytotoxicity against human cervical cancer HeLa cells. Compounds 1 and 3 significantly inhibited the proliferation of HeLa cells with IC values being 3.02 ± 0.54 and 7.16 ± 0.62 μmol·L, respectively.
Subject(s)
Humans , Alkaloids , Chemistry , Pharmacology , Cell Proliferation , HeLa Cells , Pinellia , Chemistry , Plant Extracts , Chemistry , Pharmacology , Plant Stems , Chemistry , Plant Tubers , ChemistryABSTRACT
To investigate the mechanism of lectin from Pinellia pedatisecta(PPL) on macrophage-induced inflammation and its association with inflammatory corpuscles NLRP3. Lectin from P. pedatisecta was isolated and purified by gel chromatography, and its purity was analyzed by using SDS-PAGE gel electrophoresis. ELISA was used to investigate the effect of PPL on inflammatory cytokines released by macrophages, with IL-1β as indicators;and fluorescence probe DCFH-DA fluorometer was used to determine changes in active oxygen ROS of macrophages after application of lectin from P. pedatisecta.RAW264.7 cells were pre-treated with ROS inhibitor N-acetylcysteine (NAC) to investigate the effect on ROS and the release of inflammatory factor IL-1β from macrophages to research the relationship between them. The protein levels of NLRP3, Caspase-1 p20, ASC and TXNIP were determined by Western blot.The results showed that isolated and purified PPL could reach electrophoretic purity; PPL stimulated macrophages and induced the excessive release of ROS, leading to strong oxidative stress reaction, and the levels of intracellular inflammatory factorsIL-1β were significantly increased. NAC could inhibit PPL-induced ROS excessive production and significantly reduce the release of IL-1β. In addition, PPL could induce the increase in protein expression levels of Caspase-1 p20, NLRP3 and ASC, and significantly reduce TXNIP expression. The results showed that PPL could cause a strong oxidative stress response by stimulating macrophages, activate inflammatory corpuscles NLRP3, and result in large amount of IL-1β release. That is, PPL could lead to inflammatory cascade reaction by promoting the maturation and secretion of IL-1β through ROS-TXNIP-NLRP3-IL-1β signaling pathway.
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This study was to investigate the mechanism of gingerols antagonizing the inflammatory effect of toxic raphides from Pinella pedatisecta. Mice peritonitis models induced by toxic raphides from P. pedatisecta were applied to observe the effect of gingerols on inflammatory mediators PGE2 in the exudates of abdominal inflammation in mice; rats peritoneal macrophage in vitro culture models were adopted to study the anti-inflammatory effects of gingerol against toxic raphides, with TNF-α and IL-1β in supernatant as indexes. Scanning electron microscopy was used to observe the changes in surface morphology of macrophages treated by raphides and gingerols. Macrophages-neutrophils co-cultured models were used to study the antagonism of gingerols against the effect of toxic raphides' stimulation on neutrophils migration. Results showed that gingerols could significantly inhibit the production of PGE2 in the exudates of abdominal inflammation induced by toxic raphides from P. pedatisecta in mice. Gingerols could significantly inhibit the toxic raphides from P. pedatisecta to induce the release of inflammatory factors, with certain dose dependence. Scanning electron microscopy showed that gingerols could significantly inhibit phagocytosis of macrophages, cytomembrane injury, and neutrophils migration induced by toxic raphides from P. pedatisecta. The results showed that the antagonism mechanism of gingerols against the toxic raphides from P. pedatisecta may be associated with inhibiting the pro-inflammatory toxicity including macrophage activation, inflammatory factors release, and neutrophils migration.
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AIM To investigate the anticonvulsive action of supercritical CO2 ethanol extract from Pinellia Pedatisecta Schott(SEE-CO2PP). METHODS The rat convulsive model was induced by penicillin localized injected in rat cortex. The effects of SEE-CO2PP on the latency of seizure and changes of convulsive behaviors were investigated. The latency of epileptiform discharge, and frequency and amplitude of highest wave in cortex and hippocampus were recorded by using RM6240C multichannel physiological signal collection and analysis recorder. At the same time, the contents of glutamic acid (Glu), aspartic acid (Asp), glycine (Gly) and γ-aminobutyric acid (GABA) in hippocampus were determined with high performance liquid chromatography. RESULTS SEE-CO2PP 15 and 30 g·kg-1, ig, prolonged the latent period of seizure and weakened the extent. SEE-CO2PP also prolonged the latent period of epileptiform discharge, reduced the frequency and decreased amplitude of the highest wave in both cortex and hippocampus. Moreover, SEE-CO2PP increased the content of GABA in hippocampus, but the levels of Gly,Asp and Glu had no obvious changes. CONCLUSION SEE-CO2PP inhibits the epileptiform discharge and convulsive behaviors of convulsive model rats, which suggests SEE-CO2PP has anticonvulsive action.
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Pinellia ternate and Pinellia pedatisecta lectins with biological functions could be obtained through procaryon expression.Their biological similarities and differences were analyzed and further experiments were conducted to discuss the agglutinative activity difference between polymer lectin and metamer lectin.The results showed that agglutinative activity of Pinellia pedatisecta was four times more than that of Pinellia ternate.The differences of the agglutinative activity and pharmacologic action were mainly because two site mutations of Pinellia pedatisecta in the third active site compared with the amino acids sequence in Pinellia ternate.In addition,the metamer lectin expressed by prokaryotic expression system did not agglutinate rabbit red blood cells.It conduct further discussion about the difference between two lectins and the difference between polymer lectin and metamer lectin.It would be applicable for solving the problem of lacking Pinellia resources.