RESUMEN
Variations in herb dosage due to species adulteration and dosing inaccuracies can substantially affect clinical safety and efficacy. Accurate species quantification remains challenging, as current methods often yield inconsistent results. This study introduces a novel pyrosequencing-based technique, termed herb molecular quantification (Herb-Q), designed to precisely quantify herbal products. We evaluated its effectiveness using Pinellia ternata and five of its adulterants. Initially, we assessed commonly used DNA barcodes with sequences from a public database, identifying two candidate regions, Maturase K (matK) and internal transcribed spacer 2 (ITS2), for screening specific single nucleotide polymorphism (SNP) loci, allowing for species-specific identification. These loci were validated by amplifying and sequencing genomic material from collected samples. Our validation studies showed that Herb-Q demonstrated excellent linearity, accuracy, repeatability, and detection limits. We established quantitative standard curves with high R2 values (> 0.99) to enable precise species quantification, which were combined with external standards to provide clear and accurate visual quantification results. The average bias in quantifying the tuber of P. ternata was 2.38%, confirming that Herb-Q can accurately identify and quantify herbal product constituents. Moreover, the entire quantification process took less than 4 h. This study presents a novel, rapid method for accurately quantifying species in herbal products and advances the application of DNA barcoding from species identification to quantitative detection.
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Código de Barras del ADN Taxonómico , Pinellia , Pinellia/genética , Pinellia/química , Código de Barras del ADN Taxonómico/métodos , Polimorfismo de Nucleótido Simple , ADN de Plantas/genética , Análisis de Secuencia de ADN/métodos , Medicamentos Herbarios Chinos/química , Contaminación de Medicamentos , Plantas Medicinales/genética , Plantas Medicinales/química , Plantas Medicinales/clasificaciónRESUMEN
ETHNOPHARMACOLOGICAL RELEVANCE: Chinese herb pairs are the most basic and compressed examples of Chinese herbal combinations and can be used to effectively explain the fundamental concepts of traditional Chinese medicine prescriptions. These pairings have gained significant interest due to their subtle therapeutic benefits, minimal side effects, and efficacy in treating complicated chronic conditions. The Banxia-Xiakucao Chinese herb pair (BXHP) consists of Pinellia ternata (Thunb.) Breit. (Banxia) and Prunella vulgaris L. (Xiakucao). This formula was documented in The Medical Classic of the Yellow Emperor approximately 2000 years agoï¼and clinical research has demonstrated that BXHP effectively treats insomnia. AIM OF THE STUDY: This study aimed to evaluate the efficacy and therapeutic mechanism of the BXHP through a comprehensive strategy involving network pharmacology, molecular docking, transcriptomics, and molecular biology experimental validation. MATERIALS AND METHODS: The composition of BXHP was characterized using the UPLC-Q-TOF-MS. The active compounds were screened to find drug-likeness compounds by analyzing the ADME data. To predict the molecular mechanism of BXHP in sleep deprivation (SD) by network pharmacology and molecular docking. We established a rat model of SD and the in vivo efficacy of BXHP was verified through the pentobarbital sodium righting reflex test, behavioral assays, enzyme-linked immunosorbent assay, transmission electron microscopy, HE staining, and Nissl staining, and the underlying molecular mechanism of BXHP in SD was revealed through transcriptomic and bioinformatic analyses in conjunction with quantitative real-time PCR, Western blot, and immunofluorescence staining. RESULTS: In the present study, we showed for the first time that BXHP reduced sleep latency, prolongs sleep duration, and improves anxiety; lowered serum CORT, IL6, TNF-α and MDA levels; decreased hypothalamic Glu levels; and elevated hypothalamic GABA and 5-HT levels in SD rats. We found 16 active compounds that acted on 583 targets, 145 of which are related to SD. By modularly dissecting the PPI network, we discovered three critical targets, Akt1, CREB1, and PRKACA, all of which play important roles in the effects of BXHP on SD. Molecular docking resulted in the identification of 16 active compounds that strongly bind to key targets. The results of GO and KEGG enrichment analyses of network pharmacology and transcriptomics focused on both the regulation of circadian rhythm and the cAMP signaling pathway, which strongly demonstrated that BXHP affects SD via the cAMP-PKA-CREB-Circadian rhythm pathway. Molecular biology experiments verified this hypothesis. Following BXHP administration, PKA and CREB phosphorylation levels were elevated in SD rats, the cAMP-PKA-CREB signaling pathway was activated, the expression levels of the biological clock genes CLOCK, p-BMAL1/BMAL1, and PER3 were increased, and the rhythmicity of the biological clock was improved. CONCLUSIONS: The active compounds in BXHP can activate the cAMP-PKA-CREB-Circadian rhythm pathway, improve the rhythmicity of the biological clock, promote sleep and ameliorate anxiety, which suggests that BXHP improves SD through a multicomponent, multitarget, multipathway mechanism. This study is important for the development of herbal medicines and clinical therapies for improving sleep deprivation.
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Medicamentos Herbarios Chinos , Simulación del Acoplamiento Molecular , Farmacología en Red , Pinellia , Ratas Sprague-Dawley , Privación de Sueño , Transcriptoma , Animales , Privación de Sueño/tratamiento farmacológico , Privación de Sueño/metabolismo , Masculino , Medicamentos Herbarios Chinos/farmacología , Medicamentos Herbarios Chinos/uso terapéutico , Medicamentos Herbarios Chinos/química , Ratas , Pinellia/química , Transcriptoma/efectos de los fármacos , Proteína de Unión a Elemento de Respuesta al AMP Cíclico/metabolismo , Proteína de Unión a Elemento de Respuesta al AMP Cíclico/genética , Sueño/efectos de los fármacos , Pentobarbital/farmacologíaRESUMEN
The present study investigates the chemical composition variances among Pinelliae Rhizoma, a widely used Chinese herbal medicine, and its common adulterants including Typhonium flagelliforme, Arisaema erubescens, and Pinellia pedatisecta. Utilizing the non-targeted metabolomics technique of employing UHPLC-Q-Orbitrap HRMS, this research aims to comprehensively delineate the metabolic profiles of Pinelliae Rhizoma and its adulterants. Multivariate statistical methods including PCA and OPLS-DA are employed for the identification of differential metabolites. Volcano plot analysis is utilized to discern upregulated and downregulated compounds. KEGG pathway analysis is conducted to elucidate the differences in metabolic pathways associated with these compounds, and significant pathway enrichment analysis is performed. A total of 769 compounds are identified through metabolomics analysis, with alkaloids being predominant, followed by lipids and lipid molecules. Significant differential metabolites were screened out based on VIP > 1 and p-value < 0.05 criteria, followed by KEGG enrichment analysis of these differential metabolites. Differential metabolites between Pinelliae Rhizoma and Typhonium flagelliforme, as well as between Pinelliae Rhizoma and Pinellia pedatisecta, are significantly enriched in the biosynthesis of amino acids and protein digestion and absorption pathways. Differential metabolites between Pinelliae Rhizoma and Arisaema erubescens are mainly enriched in tyrosine metabolism and phenylalanine metabolism pathways. These findings aim to provide valuable data support and theoretical references for further research on the pharmacological substances, resource development and utilization, and quality control of Pinelliae Rhizoma.
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Metabolómica , Pinellia , Rizoma , Cromatografía Líquida de Alta Presión/métodos , Metabolómica/métodos , Pinellia/metabolismo , Pinellia/química , Rizoma/metabolismo , Rizoma/química , Medicamentos Herbarios Chinos/química , Medicamentos Herbarios Chinos/metabolismo , Espectrometría de Masas/métodos , Contaminación de Medicamentos , Metaboloma , Redes y Vías MetabólicasRESUMEN
Polysaccharides from Pinelliae Rhizoma Praeparatum Cum Alumine (PPA) have various biological activities, but their properties after oral administration are not clear. In this study, the absorption, digestion and fermentation properties of PPA were studied using in vivo fluorescence tracking, in vitro simulated digestion and fecal fermentation experiments. The absorption experiment showed that fluorescence was only observed in the gastrointestinal system, indicating that PPA could not be absorbed. Simulated digestion results showed that there were no significant changes in the molecular weight, Fourier transform infrared spectroscopy (FT-IR) spectrum, monosaccharides and reducing sugar of PPA during the digestion process, showing that the overall structure of PPA was not damaged. However, the carbohydrate gel electrophoresis bands of PPA enzymatic hydrolysates after simulated digestion were significantly changed, indicating that simulated digestion might impact the configuration of PPA. In vitro fermentation showed that PPA could be degraded by microorganisms to produce short chain fatty acids, leading to a decrease in pH value. PPA can promote the proliferation of Bacteroideaceae, Megasphaera, Bacteroideaceae, and Bifidobacteriaceae, and inhibit the growth of Desulfobacteriota and Enterobacteriaceae. The results indicated that PPA could treat diseases by regulating gut microbiota, providing a scientific basis for the application and development of PPA.
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Digestión , Heces , Fermentación , Microbioma Gastrointestinal , Polisacáridos , Microbioma Gastrointestinal/efectos de los fármacos , Polisacáridos/química , Polisacáridos/farmacología , Humanos , Heces/microbiología , Heces/química , Digestión/efectos de los fármacos , Pinellia/química , AnimalesRESUMEN
ETHNOPHARMACOLOGICAL RELEVANCE: The combination of Aconitum carmichaelii Debx (Chuanwu, CW) and Pinellia ternata (Thunb.) Breit (Banxia, BX) forms an herbal pair within the eighteen incompatible medicaments (EIM), indicating that BX and CW are incompatible. However, the scientific understanding of this incompatibility mechanism, especially the corresponding drug-drug interaction (DDI), remains complex and unclear. AIM OF THE STUDY: This study aims to explain the DDI and potential incompatibility mechanism between CW and BX based on pharmacokinetics and cocktail approach. MATERIALS AND METHODS: Ultraperformance liquid chromatography-tandem mass spectrometry methods were established for pharmacokinetics and cocktail studies. To explore the DDI between BX and CW, in the pharmacokinetics study, 10 compounds were determined in rat plasma after administering CW and BX-CW herbal pair extracts. In the cocktail assay, the pharmacokinetic parameters of five probe substrates were utilized to assess the influence of BX on cytochrome P450 (CYP) isoenzyme (dapsone for CYP3A4, phenacetin for CYP1A2, dextromethorphan for CYP2D6, tolbutamide for CYP2C9, and omeprazole for CYP2C19). Finally, the DDI and incompatibility mechanism of CW and BX were integrated to explain the rationality of EIM theory. RESULTS: BX not only enhances the absorption of aconitine and benzoylaconine but also accelerates the metabolism of mesaconitine, benzoylmesaconine, songorine, and fuziline. Moreover, BX affects the activity of CYP enzymes, which regulate the metabolism of toxic compounds. CONCLUSIONS: BX altered the activity of CYP enzymes, consequently affecting the metabolism of toxic compounds from CW. This incompatibility mechanism may be related to the increased absorption of these toxic compounds in vivo.
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Aconitum , Interacciones de Hierba-Droga , Pinellia , Ratas Sprague-Dawley , Aconitum/química , Pinellia/química , Animales , Masculino , Ratas , Sistema Enzimático del Citocromo P-450/metabolismo , Espectrometría de Masas en Tándem , Extractos Vegetales/farmacocinética , Extractos Vegetales/farmacología , Extractos Vegetales/química , Medicamentos Herbarios Chinos/farmacocinética , Medicamentos Herbarios Chinos/química , Interacciones FarmacológicasRESUMEN
BACKGROUND: Pinellia pedatisecta Schott extract (PE) is extracted from Pinellia pedatisecta Schott (PPS), a traditional Chinese medicinal plant with the potential for direct anticancer effects or eliciting an anti-tumor response by activating the immune system. PURPOSE: To explore PE's ability and mechanism to reconstruct cisplatin's immunogenicity. METHODS: Cervical cancer cells were treated with cisplatin (CDDP) and/or PE. The exposure of calreticulin (CRT) on cell membrane was investigated by flow cytometry. The extracellular of ATP and HMGB1 was investigated by Western blot analysis, immunofluorescence and ELISA assay. Changes in immune profiles were using flow cytometry in vaccination and anti-tumor assays in vivo. Lastly, the mechanism of PE influenced the ROS/ERS pathway was examined by ROS assay kit, flow cytometry and Western blotting. RESULTS: PE treatment induced translocation of CRT from the endoplasmic reticulum to the cell membrane of tumor cells, concomitantly triggering immunogenic cell death (ICD). In terms of mechanisms, endoplasmic reticulum (ER) stress relievers could impede the ability of PE to induce immunogenicity. This indicates that PE is activated by ER stress, leading to subsequent induction of ICD. Upon analyzing RNA-seq data, it was observed that PE primarily induces programmed cell death in tumors by impeding upstream antioxidant mechanisms. Additionally, it transforms dying tumor cells into vaccines, activating a series of immune responses. CONCLUSIONS: This study observed for the first time that PE-induced CRT exposure on the membrane of cervical cancer cells compensates for the defect of nonimmunogenic cell death inducer CDDP thereby stimulating potent ICD. This ability restores the immunogenicity of CDDP through ER stress induced by the ROS signal. ROS played a role in PE's ability to induce ICD, leading to increased expression of ER stress-related proteins, including ATF3 and IRE-1α. PE exerted anti-cancer effects by increasing the ROS levels, and ROS/ERS signaling may be a potential avenue for cervical cancer treatment. Hence, the synergistic use of PE and CDDP holds potential for enhancing immunochemotherapy in cancer treatment.
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Calreticulina , Cisplatino , Estrés del Retículo Endoplásmico , Muerte Celular Inmunogénica , Pinellia , Especies Reactivas de Oxígeno , Neoplasias del Cuello Uterino , Cisplatino/farmacología , Neoplasias del Cuello Uterino/tratamiento farmacológico , Femenino , Pinellia/química , Estrés del Retículo Endoplásmico/efectos de los fármacos , Humanos , Muerte Celular Inmunogénica/efectos de los fármacos , Especies Reactivas de Oxígeno/metabolismo , Animales , Extractos Vegetales/farmacología , Proteína HMGB1/metabolismo , Ratones , Línea Celular Tumoral , Ratones Endogámicos BALB C , Células HeLa , Antineoplásicos/farmacologíaRESUMEN
Pinellia tuber, the dried tuber of Pinellia ternata, causes a very strong acridity sensation in the oral and laryngopharynx mucosa when taken orally in its unprocessed form. In traditional Chinese medicine (TCM), this sensation has been called "toxicity", and Pinellia tuber must be processed using ginger extract, licorice, or alum. In Japanese traditional Kampo medicine, since "toxicity" can be eliminated by decocting, it should not be processed. However, little is known about the mechanism underlying the "detoxification" of Pinellia tubers. In this study, we produced murine antiserum using recombinant P. ternata lectin (PTL), developed an immuno-fluorescence staining method for PTL in the needle-shaped crystals (raphides) that were prepared by petroleum ether extraction (PEX) from Pinellia tuber, and elucidated the mechanism of the processing of Pinellia tuber using heat or ginger extract. After heating the raphides in water, the amount of PTL contained in the raphides was significantly reduced by the immunostaining, although the shape of the raphides was not changed. Incubating raphides with dried ginger extract also significantly reduced the amount of PTL in the raphides in a concentration-dependent manner. By the activity-guided fractionation of ginger extract, the active ingredients in the ginger extract were oxalic acid, tartaric acid, malic acid, and citric acid. Among these four organic acids, oxalic acid mainly contributed to the effect of dried ginger extract by its content in ginger extract and its activity. These results exhibit scientific evidences for the traditional theories of processing to "detoxify" Pinellia tuber in TCM and Kampo medicine.
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Pinellia , Ratones , Animales , Pinellia/química , Calefacción , Extractos Vegetales/farmacología , Extractos Vegetales/química , Lectinas , Ácido OxálicoRESUMEN
ETHNOPHARMACOLOGICAL RELEVANCE: Pinellia ternata (Thunb.) Breit. (PT) has been demonstrated to be effective against the allergic airway inflammation (AAI) in clinical practices, especially in cold asthma (CA). Until now, the active ingredients, protective effect, and possible mechanism of PT against CA remain unknown. AIM OF THE STUDY: The aim of this investigation was to examine the therapeutic impact and elucidate the underlying mechanism of PT on the AAI of CA. METHODS: The compositions of PT water extract were determined via the UPLC-Q-TOF-MS/MS. The ovalbumin (OVA) and cold-water baths were used to induce CA in female mice. Morphological characteristic observations, expectorant effect, bronchial hyperreactivity (BHR), excessive mucus secretion, and inflammatory factors were used to uncover the treatment effect of PT water extract. In addition, the mucin 5AC (MUC5AC) mRNA and protein levels and the aquaporin 5 (AQP5) mRNA and protein levels were detected via qRT-PCR, immunohistochemistry (IHC), and western blotting. Moreover, the protein expressions associated with the TLR4, NF-κB, and NLRP3 signaling pathway were monitored by western blot analysis. RESULTS: Thirty-eight compounds were identified from PT water extract. PT showed significant therapeutic effects on mice with cold asthma in terms of expectorant activity, histopathological changes, airway inflammation, mucus secretion, and hyperreactivity. PT exhibited good anti-inflammatory effects in vitro and in vivo. The expression levels of MUC5AC mRNA and protein decreased significantly, while AQP5 expression levels increased significantly in the lung tissues of mice after administration with PT as compared to mice induced by CA. Furthermore, the protein expressions of TLR4, p-iκB, p-p65, IL-1ß, IL-18, NLRP3, cleaved caspase-1, and ASC were markedly reduced following PT treatment. CONCLUSIONS: PT attenuated the AAI of CA by modulating Th1- and Th2-type cytokines. PT could inhibit the TLR4-medicated NF-kB signaling pathway and activate the NLRP3 inflammasome to reduce CA. This study provides an alternative therapeutic agent of the AAI of CA after administration with PT.
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Asma , Pinellia , Femenino , Ratones , Animales , FN-kappa B/metabolismo , Proteína con Dominio Pirina 3 de la Familia NLR/metabolismo , Pinellia/química , Receptor Toll-Like 4/metabolismo , Expectorantes/uso terapéutico , Espectrometría de Masas en Tándem , Asma/patología , Transducción de Señal , Pulmón , Inflamación/patología , ARN Mensajero/metabolismo , Ovalbúmina/farmacologíaRESUMEN
The dried tuber of Pinellia ternata (Thunb.) Breit, Pinelliae Rhizoma (PR, also named 'Banxia' in Chinese), is widely used in traditional medicine. This review aims to provide detail summary of active ingredients, pharmacological effects, toxic ingredients, detoxification strategies, and omic researches, etc. Pharmacological ingredients from PR are mainly classified into six categories: alkaloids, amino acids, polysaccharides, phenylpropanoids, essential oils, and glucocerebrosides. Diversity of chemical composition determines the broad-spectrum efficacy and gives a foundation for the comprehensive utilization of P. ternata germplasm resources. The pharmacological compounds are involved in inhibition of cancer cells by targeting various pathways, including activation of immune system, inhibition of proliferation and cycle, induction of apoptosis, and inhibition of angiogenesis. The pharmacological components of PR act on nervous system by targeting neurotransmitters, activating immune system, decreasing apoptosis, and increasing redox system. Lectins, one major class of the toxic ingredients extracted from raw PR, possess significant toxic effects on human cells. Inflammatory factors, cytochrome P450 proteins (CYP) family enzymes, mammalian target of rapamycin (mTOR) signaling factors, transforming growth factor-ß (TGF-ß) signaling factors, and nervous system, are considered to be the target sites of lectins. Recently, omic analysis is widely applied in Pinellia genus studies. Plastome genome-based molecular markers are deeply used for identifying and resolving phylogeny of Pinellia genus plants. Various omic works revealed and functional identified a series of environmental stress responsive factors and active component biosynthesis-related genes. Our review summarizes the recent progress in active and toxic ingredient evaluation, pharmacological effects, detoxification strategies, and functional gene identification and accelerates efficient utilization of this traditional herb.
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Alcaloides , Medicamentos Herbarios Chinos , Pinellia , Humanos , Medicamentos Herbarios Chinos/farmacología , Pinellia/química , Multiómica , LectinasRESUMEN
Butylated hydroxytoluene (BHT), as an emerging contaminant in ecosystems, has potential influences on animals, aquatic organisms, and public health, and has been proven to be a major allelochemical of Pinellia ternata. In this study, Bacillus cereus WL08 was used to rapidly degrade BHT in liquid culture. Strain WL08 immobilized on tobacco stem charcoal (TSC) particles notably accelerated BHT removal in contract to its free cells, and exhibited excellent reutilization and storage capacities. The optimal removal parameters of TSC WL08 were ascertained to be pH 7.0, 30 °C, 50 mg L-1 BHT and 0.14 mg L-1 TSC WL08. Moreover, TSC WL08 significantly accelerated the degradation of 50 mg L-1 BHT in sterile and non-sterile soils compared to that of free WL08 or natural dissipation, and notably shortened their half-lives by 2.47- or 362.14- fold, and 2.20- or 14.99- fold, respectively. Simultaneously, TSC WL08 was introduced into the continuous cropping soils of P. ternata, which accelerated the elimination of allelochemical BHT, and notably enhanced the photosynthesis, growth, yield, and quality of P. ternata. This study provides new insights and strategies for the rapid in situ remediation of BHT-polluted soils and effective alleviation of P. ternata cropping obstacles.
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Pinellia , Suelo , Animales , Suelo/química , Hidroxitolueno Butilado/metabolismo , Bacillus cereus , Pinellia/química , Pinellia/metabolismo , Carbón Orgánico/metabolismo , Nicotiana , EcosistemaRESUMEN
To use bioinformatics and network analysis to reveal the mechanism of "Rhizoma Pinelliae-Rhizoma Coptidis" herb pair in the treatment of lung adenocarcinoma. The target and pathway of "Rhizoma Pinelliae-Rhizoma Coptidis" herb pair in the treatment of lung adenocarcinoma were explored by online databases and network analysis tools, and the potential biomarkers of "Rhizoma Pinelliae-Rhizoma Coptidis" herb pair in the treatment of lung adenocarcinoma were predicted in reverse. A total of 59 traditional Chinese medicine compounds and 510 drug targets were screened in this study. A total of 25 micro-RNAs and 15,323 disease targets were obtained through GEO2R software analysis. In the end, 294 therapeutic targets and 47 core targets were obtained. A total of 186 gene ontology enrichment assays were obtained, and core therapeutic targets play multiple roles in biological processes, molecular functions, and cellular composition. Kyoto encyclopedia of genes and genomes pathway enrichment analysis showed that the core targets were mainly enriched in cancer-related pathways, immune-related pathways, endocrine-related pathways, etc, among which the non-small cell lung cancer pathway was the most significant core pathway. Molecular docking shows that the compound and the target have good binding ability. "Rhizoma Pinelliae-Rhizoma Coptidis" herb pair plays a mechanism of action in the treatment of lung adenocarcinoma through multiple targets and pathways. miR-5703, miR-3125, miR-652-5P, and miR-513c-5p may be new biomarkers for the treatment of lung adenocarcinoma.
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Adenocarcinoma del Pulmón , Carcinoma de Pulmón de Células no Pequeñas , Medicamentos Herbarios Chinos , Neoplasias Pulmonares , MicroARNs , Pinellia , Humanos , Medicamentos Herbarios Chinos/farmacología , Pinellia/química , Simulación del Acoplamiento Molecular , Neoplasias Pulmonares/tratamiento farmacológico , Neoplasias Pulmonares/genética , Adenocarcinoma del Pulmón/tratamiento farmacológico , Adenocarcinoma del Pulmón/genéticaRESUMEN
Brassinolide (BR) is the "sixth class" plant hormone, which plays an important role in various physiological and biochemical processes of plants. The wide variety of functions of Pinellia ternata means that there is huge demand for it and thus it is in short supply. This paper mainly assessed the changes of yield and quality in P. ternata at different stages after BR treatments by principal component analysis, in order to improve the yield and quality of P. ternata and at the same time determine the best harvest time. The results showed that the tuber yield of P. ternata was significantly increased by BR treatments at different stages (except for the 15th day). After the 15th, 45th, 60th, 75th, 90th, and 105th day of treatments, the tuber yield of P. ternata reached peak values at 0.10 (0.65 g), 0.50 (1.97 g), 0.50 (1.98 g), 1.00 (2.37 g), 1.00 (2.84 g), and 2.00 mg/L (3.76 g) BR treatment, respectively. The optimal harvest time was the 75th day after 0.10, 0.50, and 1.00 mg/L BR treatments, which not only significantly improved the yield of P. ternata, but also retained high level of total alkaloids in the tubers (20.89, 5.37, and 13.44%) and bulbils (9.74, 20.42, and 13.62%), high total flavone content in the tubers (17.66, 16.26, and 12.74%) and bulbils (52.63, 12.79, and 38.69%), and high ß-sitosterol content in the tubers (25.26, 16.65, and 0.62%) of P. ternata, compared with the control, respectively.
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Alcaloides , Pinellia , Pinellia/química , Análisis de Componente Principal , Tubérculos de la Planta/química , Reguladores del Crecimiento de las Plantas/análisis , Alcaloides/análisisRESUMEN
BACKGROUND: Pinelliae Rhizoma (PR), a toxic medication, with long history, is commonly used for eliminating phlegm. Due to the shortage of wild resources and the relative lacking of cultivation technology, it is often confused with its counterfeit species in the market, such as Typhonii Rhizoma (TR), Arisaematis Rhizoma (AR) and tubers of Typhonium flagelliforme (TF) and Pinellia pedatisecta (PP). PURPOSE: It was aimed to screen signature enzymatic peptides from toxic proteins to identify PR and its four counterfeit species. STUDY DESIGN: A comparative proteogenomics strategy based on open-source transcriptome data was applied for screening signature peptides from toxic proteins, which were applied for species authentication of PR and its counterfeit species. METHODS: Firstly, the open-source transcriptome data was used for constructing the annotated protein database, which was used for peptides identification. Secondly, the toxicity of different fractions of PR were evaluated by the rat peritoneal inflammation model. Furthermore, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) were used to profile the main proteins bands of five species, whose sequences were identified based on the in-gel digestion experiment by using ultra-high-performance liquid chromatography/quadrupole-Orbitrap mass spectrometry. Finally, the label-free proteomic analysis was performed to character the proteins and screen the signature peptides of five species, which were validated in commercially available products by dynamic multi reaction monitor (DMRM). RESULTS: The results in this study confirmed that protein was the main toxic components of PR. Both Pinellia ternata agglutinin (PTA) and trypsin inhibitor (TI) like proteins are the main proteins, which were characterized by proteomic analysis based on four annotated protein database. Meanwhile, seven signature peptides from toxic proteins were screened and validated with good repeatability and specificity in commercial products. CONCLUSION: Seven signature enzymatic peptides from toxic protein screened by the comparative proteogenomics strategy based on open-source transcriptome data achieved good identification ability of PR and its four counterfeit species.
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Medicamentos Herbarios Chinos , Pinellia , Aglutininas , Animales , Medicamentos Herbarios Chinos/farmacología , Péptidos , Pinellia/química , Proteómica , Ratas , Dodecil Sulfato de Sodio , Inhibidores de TripsinaRESUMEN
ETHNOPHARMACOLOGICAL RELEVANCE: The rhizome of Pinellia ternata (Thunb.) Breit, called Pinelliae Rhizoma (PR) and Banxia in Chinese, is a well-known traditional Chinese medicine (TCM) with the functions of "removing dampness-phlegm" and "downbear counterflow and check vomiting". PR has potential toxic effects that can be detoxified by Fuzhi processing (repeated processing using one or multiple adjuvants) with specific adjuvants. AIM OF THE STUDY: This paper aims to provide a summary of traditional and current processing methods used to detoxify PR. MATERIALS AND METHODS: The available references of the processing methods of PR from the classic books of Materia Medica, literature, online databases and masters or doctoral theses are collected and summarized. We also discussed the possible processing mechanisms of how we can achieve a safer and effective application of PR via these processing methods. RESULTS: PR cannot be administered orally before processing. PR contains nucleoside alkaloids, cerebrosides, fatty acids, lectin, polysaccharides, and calcium oxalate crystals. To date, although the active substances of PR are still unclear, the toxic components are almost completely clarified as needle-like calcium oxalate crystals (NCOCs) and lectin proteins. Furthermore, the toxic effects of PR include causing death in animals, inflammation, conjunctival irritation, pregnancy toxicity, teratogenicity, visceral toxicity, aphonia and vomiting. From ancient times to now, Fuzhi methods have remained the predominant method for PR processing, and the main adjuvants used are ginger juice, alum, licorice and lime. In addition, detoxification mechanisms are related to removing or damaging the NCOC and lectin in PR based on processing with adjuvants. Currently, Fuzhi processing has been greatly improved, and novel processing technologies with novel adjuvants have been used for PR processing. However, there are still some flaws in PR processing, which should be urgently solved in the future, and clarifying the characteristic bioactive compounds in PR corresponding to its function or effects is the most important step for PR processing. CONCLUSION: Our present paper reviewed the previous literature regarding all aspects of the processing of PR, and this paper will be helpful for achieving a safer and effective application of PR and its processed products and will also be beneficial for the further optimization of processing technology and clinical medication safety of PR.
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Alcaloides , Medicamentos Herbarios Chinos , Materia Medica , Pinellia , Adyuvantes Farmacéuticos , Alcaloides/análisis , Animales , Oxalato de Calcio , Cerebrósidos/análisis , Medicamentos Herbarios Chinos/química , Ácidos Grasos/análisis , Lectinas/análisis , Materia Medica/análisis , Medicina Tradicional China , Nucleósidos/análisis , Pinellia/química , Rizoma/química , Tecnología Farmacéutica/métodos , VómitosRESUMEN
ETHNOPHARMACOLOGICAL RELEVANCE: Pinellia ternata tuber (PTT), the dried tuber of Pinellia ternata (Thunb.) Breit., has a long history of use in traditional Chinese medicine for drying dampness, resolving phlegm, down-bearing counterflow to check vomiting and dissipating masses. Modern pharmacology studies have revealed that PTT has diverse pharmacological effects such as antitussive and expectorant, anti-emetic, anti-tumor, and anti-inflammatory effect, etc. AIM OF THE REVIEW: This review aims to provide a critical and comprehensive evaluation on ethnopharmacological uses, chemical constituents, pharmacological and toxicological effects, analytical methods and quality control of PTT, which would provide scientific evidence for exploring future therapeutic, and formulating quality and safety criteria of PTT. MATERIALS AND METHODS: Pertinent information was systematically collected from several electronic scientific databases including Web of Science, Science Direct, PubMed, Elsevier, Wiley Online Library and China national knowledge infrastructure (CNKI), as well as the classic Chinese medical books. RESULTS: PTT is reported to be widely used traditionally for the treatment of cough, vomiting, infection, and inflammatory diseases in many southeast Asian countries. Phytochemical studies have revealed the presence of a total of 233 compounds belonging to alkaloids, nucleosides, organic acids, polysaccharides, volatile oils, amino acids, proteins, starches, etc. The extracts and components of PTT have possessed diverse pharmacological activities, such as antitussive, antiemetic, antitumor, antibacterial, and sedative-hypnotic activities. Raw P. ternata tuber (RPTT) with a pungent taste causes acrid irritation of the oral and laryngopharynx mucosa when taken by mistake, while its toxicity and side effects of RPTT can be dramatically reduced with proper processing. Three kinds of processed P. ternata tuber with different processing methods are available and traded in market, as well as applied in clinical treatments. Additionally, although raw or processed PTT have been recorded in several mainstream pharmacopoeias such as Chinese Pharmacopoeia, Japanese Pharmacopoeia, and Korean Pharmacopoeia, the quality items and requirements varies a lot. Therefore, a unified international standard of raw and processed PTT is urgent need to be done. CONCLUSIONS: The ethnopharmacological, phytochemical, pharmacological and toxicological and quality evaluation of PTT were highlighted in this review, which provides potential reference information to future investigate and commercially explore for pharmaceutical applications. Nevertheless, an efficient method for chemical profiling is still unavailable to find potent bioactive markers for quality control, and then comprehensive pharmacological effects and mechanisms and toxicological evaluation of PTT require further detailed research to ensure their quality and safety.
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Antitusígenos , Pinellia , Etnofarmacología , Fitoquímicos/uso terapéutico , Fitoquímicos/toxicidad , Pinellia/química , Control de Calidad , VómitosRESUMEN
ETHNOPHARMACOLOGICAL RELEVANCE: Pinelliae Rhizoma Praeparatum (PRP) is a traditional processed product of Pinellia ternata (Thunb.) Berit., which mainly used for treating cold asthma (CA). However, the mechanism of action of PRP for treating CA have not been fully elucidated. AIM OF THE STUDY: To investigate the core active constituents and the pharmacological mechanism of PRP against CA. MATERIALS AND METHODS: Ovalbumin (OVA) and cold water-induced cold asthma model were established in male mice. The effects of water extract from PRP were evaluated by general morphological observation, expectorant activity, airway hyperresponsiveness, mucus hypersecretion, inflammatory cytokines, etc. Additionally, the mRNA and protein expression of mucin 5AC (MUC5AC) and aquaporin 5 (AQP5) in vivo and in vitro were detected by immunohistochemistry (IHC), qRT-PCR, and western blotting. The mechanisms of action were investigated through network pharmacology and transcriptomic, and validated through western blotting and molecular docking. RESULTS: PRP exhibited a favorable expectorant activity, and significantly reduced the airway inflammation, mucus secretion, and hyperresponsiveness in cold asthma model. It also reduced the levels of IL-4, IL-5, IL-8, and IL-13 in bronchoalveolar lavage fluid (BALF) and IL-4 and total IgE in serum, while obviously increased the levels of IL-10 and IFN-γ in serum for asthmatic mice. Meanwhile, PRP also attenuated the pathological changes and mucus production in cold asthmatic mice. Moreover, the downregulation of MUC5AC and upregulation of AQP 5 were detected by western blotting and qRT-PCR after administration with PRP both in vivo and in vitro. PRP expectedly inhibited the protein expression of PKC-α, SRC, p-EGFR, p-ERK1/2, p-JNK, p-p38, p-PI3K, and p-Akt levels in vivo. CONCLUSIONS: These combined data showed that PRP suppressed the allergic airway inflammation of CA by regulating the balance of Th1 and Th2 cytokines and the possible involvement of the PKC/EGFR/MAPK/PI3K-Akt signaling pathway. Pentadecanoic acid, licochalcone A, ß-sitosterol, etc. were considered as main active ingredients of PRP against CA. This study provides a novel perspective of the classical herbal processed product PRP in the treatment of CA.
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Asma , Pinellia , Animales , Asma/patología , Líquido del Lavado Bronquioalveolar/química , Citocinas/metabolismo , Receptores ErbB/metabolismo , Expectorantes/uso terapéutico , Inflamación/metabolismo , Interleucina-4/metabolismo , Pulmón , Sistema de Señalización de MAP Quinasas , Masculino , Ratones , Ratones Endogámicos BALB C , Simulación del Acoplamiento Molecular , Moco/metabolismo , Ovalbúmina/farmacología , Fosfatidilinositol 3-Quinasas/metabolismo , Pinellia/química , Proteínas Proto-Oncogénicas c-akt/metabolismo , Transducción de Señal , Agua/farmacologíaRESUMEN
This study was designed to identify the pathogen causing soft rot of Pinellia ternata in Qianjiang of Hubei province and screen out the effective bactericides, so as to provide a theoretical basis for the control of soft rot of P. ternata. In this study, the pathogen was identified based on molecular biology and physiological biochemistry, followed by the detection of pathogenicity and pathogenicity spectrum via plant tissue inoculation in vitro and the indoor toxicity determination using the inhibition zone method to screen out bactericide with good antibacterial effects. The control effect of the bactericide against P. ternata soft rot was verified by the leave and tuber inoculation in vitro. The phylogenetic tree was constructed based on the 16 S rDNA, dnaX gene, and recA gene sequences, respectively, and the result showed that the pathogen belonged to the same branch as the type strain Dickeya fangzhongdai JS5. The physiological and biochemical tests showed that the pathogen was identical to D. fangzhongdai, which proved that the pathogen was D. fangzhongdai. The pathogenicity test indicated that the pathogen could obviously infect leaves at 24 h and tubers in 3 d. As revealed by the indoor toxicity test, 0.3% tetramycin, 5% allicin, and 80% ethylicin had good antibacterial activities, with EC_(50) values all less than 50 mg·L~(-1). Tests in tissues in vitro showed that 5% allicin exhibited the best control effect, followed by 0.3% tetramycin and 10% zhongshengmycin oligosaccharide, and their preventive effects were better than curative effects. Therefore, 5% allicin can be used as the preferred agent for the control of P. ternata soft rot, and 0.3% tetramycin and 10% zhongshengmycin oligosaccharide as the alternatives. This study has provided a certain theoretical basis for the control of P. ternata soft rot.
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Pinellia , Filogenia , Pinellia/química , Hojas de la Planta , Tubérculos de la PlantaRESUMEN
Benzene can impair peripheral immunity and immune organs; however, the recovery of benzene impairment has rarely been reported. In this study, we developed an immune dysfunction mouse model using a benzene gavage (500 mg/kg). Female Balb/c mice were treated with Bombyx batryticatus (BB, 5 g/kg), raw pinellia (RP, 5 g/kg), or a combination of Valproic acid and Coenzyme Q10 (CM, 150 mg/kg VPA & 100 mg/kg CoQ10) medication for four weeks. The immune function of the peripheral blood mononuclear cells (PBMCs), spleen, and thymus was determined to evaluate whether the observed impairment could be altered by medications in the mouse model. Results showed that medications could alleviate benzene-induced structural and functional damage of spleen and thymus. Benzene exposure decreased the ATP level of PBMC, which can be improved by BB, RP or CM. Importantly, BB, RP or CM could relieve benzene induced-oxidative stress by increasing the activities of glutathione peroxidase (GSH) and superoxide dismutase (SOD) and decreasing the contents of malondialdehyde (MDA). In conclusion, BB, RP, and CM were able to alleviate the benzene-induced immune dysfunction and redox imbalance. Improvement of the oxidative and antioxidant imbalance may represent a mechanism by which medicine prevents benzene-induced immune dysfunction.
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Benceno/toxicidad , Inmunidad/efectos de los fármacos , Leucocitos Mononucleares/efectos de los fármacos , Bazo/efectos de los fármacos , Timo/efectos de los fármacos , Adenosina Trifosfato/sangre , Animales , Bombyx/química , Femenino , Glutatión Peroxidasa/efectos de los fármacos , Glutatión Peroxidasa/metabolismo , Leucocitos Mononucleares/metabolismo , Masculino , Ratones , Ratones Endogámicos BALB C , Pinellia/química , Extractos Vegetales/farmacología , Organismos Libres de Patógenos Específicos , Superóxido Dismutasa/efectos de los fármacos , Superóxido Dismutasa/metabolismo , Ubiquinona/farmacología , Ácido Valproico/farmacologíaRESUMEN
This study was designed to identify the pathogen causing soft rot of Pinellia ternata in Qianjiang of Hubei province and screen out the effective bactericides, so as to provide a theoretical basis for the control of soft rot of P. ternata. In this study, the pathogen was identified based on molecular biology and physiological biochemistry, followed by the detection of pathogenicity and pathogenicity spectrum via plant tissue inoculation in vitro and the indoor toxicity determination using the inhibition zone method to screen out bactericide with good antibacterial effects. The control effect of the bactericide against P. ternata soft rot was verified by the leave and tuber inoculation in vitro. The phylogenetic tree was constructed based on the 16 S rDNA, dnaX gene, and recA gene sequences, respectively, and the result showed that the pathogen belonged to the same branch as the type strain Dickeya fangzhongdai JS5. The physiological and biochemical tests showed that the pathogen was identical to D. fangzhongdai, which proved that the pathogen was D. fangzhongdai. The pathogenicity test indicated that the pathogen could obviously infect leaves at 24 h and tubers in 3 d. As revealed by the indoor toxicity test, 0.3% tetramycin, 5% allicin, and 80% ethylicin had good antibacterial activities, with EC_(50) values all less than 50 mg·L~(-1). Tests in tissues in vitro showed that 5% allicin exhibited the best control effect, followed by 0.3% tetramycin and 10% zhongshengmycin oligosaccharide, and their preventive effects were better than curative effects. Therefore, 5% allicin can be used as the preferred agent for the control of P. ternata soft rot, and 0.3% tetramycin and 10% zhongshengmycin oligosaccharide as the alternatives. This study has provided a certain theoretical basis for the control of P. ternata soft rot.
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Filogenia , Pinellia/química , Hojas de la Planta , Tubérculos de la PlantaRESUMEN
This study investigated the material basis and mechanism of Pinelliae Rhizoma Decoction in the treatment of airway inflammation. The cigarette smoke combined with lipopolysaccharide(LPS) was used to induce an airway inflammation model in mice. The expression levels of IL-6 and IL-8 in the bronchoalveolar lavage fluid(BALF) and the phosphorylation levels of p38 and IκB in the lungs of mice were taken as indexes to screen the effective extracts by system solvent extraction from Pinelliae Rhizoma Decoction(dichloromethane extract, ethyl acetate extract, n-butanol extract, etc.). Meanwhile, the human bronchial epithelial(16-HBE) cell model of cigarette smoke extract(CSE)-induced injury was established, and the mRNA expression levels of IL-6 and IL-8 and the phosphorylation levels of p38 and IκB proteins were also taken as indexes to evaluate the anti-inflammatory effect of different extracts of Pinelliae Rhizoma Decoction. The results showed that Pinelliae Rhizoma Decoction significantly antagonized airway inflammation in mice by down-regulating the expression levels of IL-6 and IL-8 in mice with airway inflammation and 16-HBE cells with CSE-induced injury and inhibiting the phosphorylation levels of p38 and IκB. The dichloromethane and ethyl acetate extracts of Pinelliae Rhizoma Decoction showed significant anti-inflammatory effects, while such effects of other extracts were not prominent. Furthermore, the database of Pinelliae Rhizoma composition was constructed, and the components in effective extracts were analyzed by HPLC-TOF-MS and Nano-LC-MS/MS. As revealed by the results, the compositions of the two effective extracts were similar with 36 common components. They were combined and then divided into Pinelliae Rhizoma alkaloids(PTAs) and Pinelliae Rhizoma non-alkaloids(PTNAs) by 732 cation-exchange resin. Further in vitro investigation confirmed the significant anti-inflammatory effect of PTNAs, while such effect of PTAs was not manifest. The MS analysis showed 172 peptides and 7 organic acids in PTNAs. The peptide content in PTNAs was 63.5% measured by quantitative analysis of BCA assay, and the organic acid content was 9.92% by potentiometric titration method. The findings of this study suggested that Pinelliae Rhizoma Decoction could antagonize airway inflammation in mice by inhibiting phosphorylation of p38 and IκB and blocking the activation of MAPK and NF-κB signaling pathways, and the effective components were related to the peptides and organic acids in PTNAs. The above results lay a foundation for the research on the mechanism and material basis of Pinelliae Rhizoma in antagonizing airway inflammation.