Pollen Typhae Research Articles

Discovery of two new long chain fatty acid esters of flavonol glycoside from Pollen Typhae Carbonisatus and their protective effects on HBMECs

ABATRACT Two new compounds, namely, isorhamnenin-3-O-neohesperidin-6′′-linoleic acid ester (1) and isorhamnenin-3-O-neohesperidin-6′′-palmitate (2), along with 17 known compounds, were isolated from Pollen Typhae Carbonisatus (PTC). Their structures were elucidated on the basis of one-dimensional (1D)-, two-dimensional (2D)-nuclear magnetic resonance (NMR), and high-resolution electrospray ionization mass spectrometry (HRESIMS) spectroscopic data analyses. The two new compounds (1 and 2) exhibited a protective effect in a dose-dependent manner and promoted tube generation in the oxygen-glucose deprivation/reoxygenation (OGD/R)-induced human brain microvascular endothelial cells (HBMECs).

Integrated transcriptomics and lipidomics reveals protective effect in vascular endothelial barrier of a polysaccharide from Typhae Pollen

Endothelial cell dysfunction caused by inflammation and even vascular leakage are important manifestations of blood stasis syndrome (BSS). Reversible regulation of vascular integrity to cure BSS has attracted considerable interest. Herein, a novel acidic polysaccharide (TPP-4) was purified and characterized from Typhae Pollen, a typical traditional Chinese medicine for treating BSS, especially for bleeding caused by blood stasis. A series of structural characterization methods, including spectroscopic methods (FT-IR and UV), chromatographic methods (HPGPC, HPAEC-PDA and GC–MS) and NMR, have been used to reveal the fine structure of TPP-4. TPP-4 was a homogeneous heteropolysaccharide comprised with RG-I backbone. TPP-4 showed fantastic activities in vascular integrity regulation both in vitro (HUVECs) and in vivo (zebrafish). Transcriptomics revealed that SOX7 and lipid metabolism were the potential targets. Lipidomics showed that TPP-4 could regulate lipid metabolism disorders caused by vascular inflammation, particularly affecting LPE levels. The above regulatory effects were furtherly demonstrated to be related with VEGFA/PI3K/mTOR signaling pathway through various molecular biological experiments.

A novel strategy for the quality control of carbonized Typhae pollen using colorimeter, liquid chromatography-mass spectrometry, and efficacy evaluation coupled with multivariate statistical analysis.

In this study, a novel quality control strategy was proposed, aiming to establish a multivariate specification for the processing step by exploring the correlation between colors, chemical components, and hemostatic effects of the carbonized Typhae pollen (CTP) using multivariate statistical analysis. The CTP samples were stir-fried at different durations. Afterward, the colorimeter and LC-MS techniques were applied to characterize the CTP samples, followed by the determination of bleeding time and clotting time using mice to evaluate their hemostatic effect. Then, principal component analysis, hierarchical cluster analysis, and multi-block partial least squares were used for data analysis on colors, chemical components, and their correlation with the hemostatic effect. Consequently, 13 critical quality attributes (CQAs) of CTP were identified via multivariate statistical analysis-L*, a*, b*, 3,4-dihydroxybenzoic acid, 4-hydroxybenzoic acid, 3-hydroxybenzoic acid, quercetin-3-O-glucoside, azelaic acid, kaempferol-3-O-glucoside, quercetin, naringenin, kaempferol, and isorhamnetin. The multivariate specification method involving the 13 CQAs was developed and visualized in the latent variable space of the partial least squares model, indicating that the proposed method was successfully applied to assess the quality of CTP and the degree of carbonization. Most importantly, this study offers a novel insight into the control of processing for carbonized Chinese herbal medicines.

Identifying the active ingredients of carbonized Typhae Pollen by spectrum-effect relationship combined with MBPLS, PLS, and SVM algorithms

Typhae Pollen (TP) and its carbonized product (carbonized Typhae Pollen, CTP), as cut-and-dried herbal drugs, have been widely used in the form of slices in clinical settings. However, the two drugs exhibit a great difference in terms of their clinical efficacy, for TP boasts an effect of removing blood stasis and promoting blood circulation, while CTP typically presents a hemostatic function. Since the active ingredients of CTP, so far, still remain unclear, this study aimed at identifying the active ingredients of CTP by spectrum-effect relationship approach coupled with multi-block partial least squares (MBPLS), partial least squares (PLS), and support vector machine (SVM) algorithms. In this study, the chemical profiles of a series of CTP samples which were stir-fried for different duration (denoted as CTP0∼CTP9) were firstly characterized by UHPLC-QE-Orbitrap MS. Then the hemostatic effect of the CTP samples was evaluated from the perspective of multiple parameters—APTT, PT, TT, FIB, TXB2, 6-keto-PGF1α, PAI-1 and t-PA—using established rat models with functional uterine bleeding. Subsequently, MBPLS, PLS and SVM were combined to perform spectrum-effect relationship analysis to identify the active ingredients of CTP, followed by an in vitro hemostatic bioactivity test for verification. As a result, a total of 77 chemical ingredients were preliminarily identified from the CTP samples, and the variations occurred in these ingredients were also analyzed during the carbonizing process. The study revealed that all the CTP samples, to a varying degree, showed a hemostatic effect, among which CTP6 and CTP7 were superior to the others in terms of the hemostatic effect. The block importance in the projection (BIP) indexes of MBPLS model indicated that flavonoids and organic acids made more contributions to the hemostatic effect of CTP in comparison to other ingredients. Consequently, 9 bioactive ingredients, including quercetin-3-O-glucoside, kaempferol-3-O-rutinoside, quercetin, kaempferol, isorhamnetin, 2-methylenebutanedioic acid, pentanedioic acid, benzoic acid and 3-hydroxybenzoic acid, were further identified as the potential active ingredients based on PLS and SVM models as well as the in vitro verification. This study successfully revealed the bioactive ingredients of CTP associated with its hemostatic effect, and also provided a scientific basis for further understanding the mechanism of TP processing. In addition, it proposed a novel path to identify the active ingredients for Chinese herbal medicines.

Ultrasmall and highly biocompatible carbon dots derived from natural plant with amelioration against acute kidney injury

BackgroundAcute kidney injury (AKI) refers to a tricky clinical disease, known by its high morbidity and mortality, with no real specific medicine for AKI. The carbonization product from Pollen Typhae (i.e., Pu-huang in China) has been extensively employed in clinic, and it is capable of relieving the renal damage and other diseases in China since acient times.ResultsInspired by the carbonization process of Traditional Chinese Medicine (TCM), a novel species of carbon dots derived from Pollen Typhae (PT-CDs) was separated and then collected using a one-pot pyrolysis method. The as-prepared PT-CDs (4.85 ± 2.06 nm) with negative charge and abundant oxygenated groups exhibited high solubility, and they were stable in water. Moreover, the rhabdomyolysis (RM)-induced AKI rat model was used, and it was first demonstrated that PT-CDs had significant activity in improving the level of BUN and CRE, urine volume and kidney index, and histopathological morphology in RM-induced AKI rats. It is noteworthy that interventions of PT-CDs significantly reduced degree of inflammatory reaction and oxidative stress, which may be correlated with the basial potential mechanism of anti-AKI activities. Furthermore, cytotoxicity assay and biosafety evaluation exhibited high biocompatibility of PT-CDs.ConclusionThis study offers a novel relieving strategy for AKI based on PT-CDs and suggests its potential to be a related candidate for clinical applications.Graphical

An arabinogalactan isolated from Pollen Typhae induces the apoptosis of RKO cells by promoting macrophage polarization

An arabinogalactan (PTPS-1-2) was isolated and characterized from Pollen Typhae, and its potential antitumor effects on activating macrophages to produce immunomodulatory factors and promoting apoptosis in colorectal cancer cells were investigated. Structural characterization showed that PTPS-1-2 had a molecular weight of 59 kDa and was composed of rhamnose, arabinose, glucuronic acid, galactose, and galacturonic acid with a molar ratio of 7.6: 17.1: 6.5: 61.4: 7.4. Its backbone was predominantly composed of T-β-D-Galp, 1,3-β-D-Galp, 1,6-β-D-Galp, 1,3,6-β-D-Galp, 1,4-α-D-GalpA, 1,2-α-L-Rhap, additionally, branches contained 1,5-α-L-Araf, T-α-L-Araf, T-β-D-4-OMe-GlcpA, T-β-D-GlcpA and T-α-L-Rhap. PTPS-1-2 activated RAW264.7 cell by triggering the NF-kB signaling pathway and M1 macrophage polarization. Furthermore, the conditioned medium (CM) of Mφ pretreated with PTPS-1-2 exerted marked antitumor effects by inhibiting RKO cell proliferation and suppressing cell colony formation. Collectively, our findings suggested that PTPS-1-2 might be a therapeutic option for the prevention and treatment of tumors.

Identification of hub genes and candidate herbal treatment in obesity through integrated bioinformatic analysis and reverse network pharmacology

Obesity is a global epidemic elevating the risk of various metabolic disorders. As there is a lack of effective drugs to treat obesity, we combined bioinformatics and reverse network pharmacology in this study to identify effective herbs to treat obesity. We identified 1011 differentially expressed genes (DEGs) of adipose tissue after weight loss by analyzing five expression profiles (GSE103766, GSE35411, GSE112307, GSE43471, and GSE35710) from the Gene Expression Omnibus (GEO) database. We identified 27 hub genes from the protein–protein interaction (PPI) network by performing MCODE using the Search Tool for the Retrieval of Interacting Genes (STRING) database. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses revealed that these hub genes have roles in the extracellular matrix–receptor interaction, cholesterol metabolism, PI3K-Akt signaling pathway, etc. Ten herbs (Aloe, Portulacae Herba, Mori Follum, Silybum Marianum, Phyllanthi Fructus, Pollen Typhae, Ginkgo Semen, Leonuri Herba, Eriobotryae Folium, and Litseae Fructus) targeting the nine hub genes (COL1A1, MMP2, MMP9, SPP1, DNMT3B, MMP7, CETP, COL1A2, and MUC1) using six ingredients were identified as the key herbs. Quercetin and (-)-epigallocatechin-3-gallate were determined to be the key ingredients. Lastly, Ingredients-Targets, Herbs-Ingredients-Targets, and Herbs-Taste-Meridian Tropism networks were constructed using Cytoscape to elucidate this complex relationship. This study could help identify promising therapeutic targets and drugs to treat obesity.

Pollen Typhae-Based Magnetic-Powered Microrobots toward Acute Gastric Bleeding Treatment.

Traditional Chinese herbal medicine (TCHM) is the naturally available pharmaceutical with millennia of evolution from ancient China, capable of a superior therapeutic index and minimized unwanted effects on the human body. This work presents a therapeutic microrobotic platform based on pollen typhae (PT), a typical type of TCHM, fabricated by coating porous PT microspheres with Fe3O4 nanoparticles (PT robots) via electrostatic adsorption. The PT robots exhibit effective and controllable motion in various biological media upon external magnetic control and, meanwhile, preserve the inherent hemostasis property of PT. The blood clotting capacity of PT robots is attributed to their stimulation of the endogenous blood coagulation pathway and platelets with increased counts, which could be further improved by their effective magnetic propulsion. The remote magnetic control also allows the manipulation of PT robots in mice stomach, inducing enhanced binding and prolonged retention of PT robots in stomach mucosa. Moreover, PT robots upon magnetic control show an enhanced hemostatic effect in treating the mice bearing acute gastric bleeding compared with other passive groups. This work offers a facile and feasible route to integrate TCHM with manmade micromachines possessing the innate curative features of TCHM. Such a design expanded the versatility of microrobots and can be generalized to vast types of TCHM for broader biomedical applications.

Discovery of processing-associated Q-marker of carbonized traditional Chinese medicine: An integrated strategy of metabolomics, systems pharmacology and in vivo high-throughput screening model

BackgroundCarbonized traditional Chinese medicine (TCM) is a kind of distinctive traditional medicine, which has been widely used to cure various bleeding syndromes in clinic for over 2000 years. However, there are no effective quality control methods developed on carbonized TCM so far. PurposeThis study aimed at developing a processing-associated quality marker (Q-marker) discovery strategy, which would enable to promote the quality control study of carbonized TCM. MethodsCarbonized Typhae Pollen (CTP), a typical carbonized TCM with fantastic efficacy of stanching bleeding and removing blood stasis, was used as an example. First, a ultraperformance liquid chromatography with quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS) method was established to characterize four types of CTP in different processing degrees. Second, chemometric method was applied to screen candidate Q-markers. Third, peak area changes and Aratio changes of each candidate markers in 57 batches samples were described (Traceability and Transitivity). Fourth, systems pharmacology and two high-throughput zebrafish models: cerebral hemorrhage model and thrombus model were used to furtherly screen Q-markers (Effectiveness). Finally, a ultraperformance liquid chromatographic coupled with triple quadrupole tandem mass spectrometry (UPLC-TQ-MS) method was established and applied to quantify Q-markers in additional 10 batches of CTP samples (Measurability). ResultsThe chemical profiles of Typhae Pollen during the carbonized process were investigated. Then, 12 candidate compounds were screened in chemometric part. Six Q-markers (isorhamnetin-3-O-neohesperidoside, isorhamnetin-3-O-rutinoside, kaempferol-3-O-neohesperidoside, naringenin, quercetin and isorhamnetin) were subsequently screened out using three principles of Q-markers combined with content changes and two in vivo zebrafish models. Their average contents in additional 10 batches of CTP were 316.8 μg/g, 13.7 μg/g, 6.1 μg/g, 197.8 μg/g, 12.9 μg/g and 199.3 μg/g, respectively. Their content proportion was about 25: 1: 0.5: 15: 1: 15. ConclusionA processing-associated Q-marker discovery strategy was developed for carbonized TCM. It might provide a novel insight to solve the problem of ‘Chao Tan Cun Xing’ in carbonized process.

Effect of Shixiao San on inflammatory factors and pain in rats with endometriosis

Ethnopharmacological relevanceIn the practice of traditional Chinese medicine, endometriosis is believed to be caused by blood stasis and is characterised by dysmenorrhea, which is difficult to control. Shixiao San (SXS) has a long history of use in the treatment of gynaecological diseases. The prescriptions composed of SXS include Typhae Pollen and Faeces Trogopterori, both of which have anti-inflammatory activity. In addition, Typhae Pollen can be used to treat many kinds of blood stasis diseases. Aim of the studyThe purpose of the present study was to investigate the effect of SXS on pain relief in rats with endometriosis and to preliminarily explore its mechanism of action in alleviating pain. Material and methodsTen rats received sham operation as the Sham group, and 30 endometriosis model rats were randomly divided into three groups: the Model, Shixiao San-Low (SXS-L), and Shixiao San-High (SXS-H) groups. The rats were administered the appropriate treatment via intragastric gavage for 4 weeks. The thermal radiation pain and mechanical pain thresholds of the rats were measured every 7 days after treatment. Finally, the distribution density of nerve fibres in endometrial tissue, the inflammatory infiltration of the dorsal root ganglion (DRG), the expression of TRPV1 in the DRG, and the expression of IL-1β, TNF-α, and IL-6 in ectopic tissue were measured. ResultsAfter SXS treatment, the growth of ectopic tissue in rats with endometriosis was significantly suppressed, their thermal radiation pain and mechanical pain thresholds increased, the density of nerve fibres and the expression of inflammatory factors in ectopic tissues reduced, and inflammatory cells infiltration in the DRG of the animals alleviated. Meanwhile, the expression of TRPV1 in the DRG was downregulated in rats with endometriosis. ConclusionsSXS could possibly inhibit the development of endometriosis and relieve pain in patients with endometriosis by reducing inflammatory responses in ectopic tissue and the DRG.

Investigating the Mechanisms of Pollen Typhae in the Treatment of Diabetic Retinopathy Based on Network Pharmacology and Molecular Docking.

Objective To explore the main bioactive compounds and investigate the underlying mechanism of Pollen Typhae (PT) against diabetic retinopathy (DR) by network pharmacology and molecular docking analysis. Methods Bioactive ingredients and the target proteins of PT were obtained from TCMSP, and the related target genes were acquired from the SwissTargetPrediction database. The target genes of DR were obtained from GeneCards, TTD database, DisGeNET database, and DrugBank. The compound-target interaction network was established based on Cytoscape 3.7.2. The protein-protein interaction (PPI) network was constructed via STRING database and Cytoscape 3.7.2. Gene ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis were visualized through DAVID database and Bioinformatics. Ingredient-gene-pathway network analysis was conducted to further screen the ingredients, target proteins, and pathways closely related to the biological mechanism on PT for DR, and molecular docking analysis was performed by SYBYL-X 2.1.1 software. Finally, the mechanism and underlying targets of PT in the treatment of DR were predicted. Results A total of 8 compounds and 171 intersection targets were obtained based on the online network database. 7 main compounds were screened from compound-target network, and 53 targets including the top six key targets (PTGS2, AKT1, VEGFA, MAPK3, TNF, and EGFR) were further acquired from PPI analysis. The 53 key targets covered 80 signaling pathways, among which PI3K-Akt signaling pathway, focal adhesion, Rap1 signaling pathway, VEGF signaling pathway, and HIF-1 signaling pathway were closely connected with the biological mechanism involved in the alleviation of DR by PT. Ingredient-gene-pathway network shows that AKTI, EGFR, and VEGFA were core genes, kaempferol and isorhamnetin were pivotal ingredients, and VEGF signaling pathway and Rap1 signaling pathway were closely involved in anti-DR. The docking results indicated that five main compounds (arachidonic acid, isorhamnetin, quercetin, kaempferol, and (2R)-5,7-dihydroxy-2-(4-hydroxyphenyl)chroman-4-one) had good binding activity with EGFR and AKT1 targets. Conclusion The active ingredients in PT may regulate the levels of inflammatory factors, suppress the oxidative stress, and inhibit the proliferation, migration, and invasion of retinal pericytes by acting on PTGS2, AKT1, VEGFA, MAPK3, TNF, and EGFR targets through VEGF signaling pathway, PI3K-Akt signaling pathway, Rap1 signaling pathway, and HIF-1 signaling pathway to play a therapeutic role in diabetic retinopathy.

Ethanolic extract of Puhuang (Pollen Typhae) modulates lipopolysaccharide-induced inflammatory response through inducible nitric oxide synthase / cyclooxygenase-2 signaling in RAW 264.7 macrophage.

To evaluate the immune modulatory response of Puhuang (Pollen Typhae), ethanolic extract of dried pollens (TP-E) and charcoal activated pollens (CTP-E) were used for their phytochemical evaluation and their modulatory response against lipopolysaccharide (LPS) induced inflammatory activity on RAW264.7 macrophage cells. Biochemical assays were carried out to quantify the 1,1-Diphenyl-2-picrylhydrazyl Radical Scavenging Activity, Reducing Power, Ferrous ion chelating ability and total polyphenol content and flavonoids. Non-toxic dose of the extract (TP-E and CTP-E) was chosen based on 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Effect of TP-E and CTP-E on lipopolysaccharides-induced inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) expression was measured by Western blot and quantitative PCR (qRT-PCR). Expression of inflammatory cytokines, such as interleukins (IL-1β and IL-6) and tumor necrosis factor α (TNF-α), was quantified using qRT-PCR. Mitogen-activated protein kinase pathway was analyzed using Western blot. Phytochemical analysis revealed that both TP-E and CTP-E have strong antioxidant activities and high flavonoid and phenolic contents. TP-E and CTP-E effectively inhibit the expression of iNOS and COX-2, thereby inhibiting its downstream proinflammatory regulators, the extracellular signal-related kinase-1/2, that decreases the expression of IL-1β, IL-6 and TNF-α. Phytochemical constituents present in Typha angustifolia Linn could be used for treating inflammation-related diseases.

A gradient-based discriminant analysis method for process quality control of carbonized TCM via Fourier transform near infrared spectroscopy: A case study on carbonized Typhae Pollen

Carbonized traditional Chinese medicine (TCM) is a kind of distinctive traditional drug which has been widely used in various bleeding syndromes for over two thousand years, and most of them are still in clinical use. Although they share similar processing method: stir-frying, there are no specific quality standards and few quality control researches carried out on carbonized TCM up until now. Carbonized Typhae Pollen (CTP) is a typical carbonized TCM with efficacy of eliminating blood stasis and stanching bleeding. In this study, a novel process quality control model coupled with near infrared spectroscopy was established, called Gradient-based Discriminant Analysis method (GDA). Compared with conventional modeling methods (Convolutional Neural Network, Linear Discriminant Analysis, Standard Normal Variate-LDA), GDA model applied in fiber optic probe acquisition mode exhibited highest test accuracy (0.961), satisfactory correct identification (internal validation, 100%; external validation, 97.1%) and excellent model stability. This method provided a perfect guideline for process quality control of Carbonized TCM as well as ensured their clinical efficacy.

The study of neuroprotective effects and underlying mechanism of Naoshuantong capsule on ischemia stroke mice

BackgroundNaoshuantong capsule (NSTC) is an oral Chinese medicine formula composed of Typhae Pollen, Radix Paeoniae Rubra, Curcumae Radix, Gastrodiae Rhizoma and Radix Rhapontici. It has been widely used at the acute and recovery stage of ischemic stroke since 2001. Comparing with its wide clinical application, there are only few studies emphasize on investigating its pharmacological effects.MethodsTo more generally elucidate the underlying mechanisms in this study, we identified active ingredients in NSTC by a network pharmacology approach based on transcriptomics analysis and pharmacological experiments. Modified neurological severity scores and morphometric analysis using Nissl staining were employed to evaluate the neuroprotective effects of NSTC on ischemia stroke in mice.ResultsThe results showed that NSTC had preventive and protective effects on ischemia stroke, featuring repair of brain tissue during the sub-acute stage of stroke. This may attribute to the underlying mechanisms including anti-inflammatory, antioxidant, and anti-apoptotic activities, as well as an attenuation of excitatory amino acids (EAAs) toxicity of the active ingredients, especially the most active apigenin, from NSTC. Specifically, naringenin, calycosin, gastrodin, caffeic acid, paeoniflorin, and β-elemene seem to be also pharmacological active substances responsible for the anti-inflammatory effects. Meanwhile, 13-hydroxygemone, gastrodin, and p-hydroxybenzyl alcohol contributed to the attenuation of EAAs toxicity Furthermore, apigenin, naringenin, calycosin, gastrodin, and β-elemene accelerated the repair of brain ischemic tissue by up-regulating the expression of TGF-β1 levels.ConclusionsThe present study identifies the active ingredients of NSTC and illustrates the underlying mechanism using a combination of network pharmacology, transcriptomics analysis, and pharmacological experiments.

Evaluation of VEGF mediated pro-angiogenic and hemostatic effects and chemical marker investigation for Typhae Pollen and its processed product

Ethnopharmacological relevanceTyphae Pollen (TP) is a well-known Traditional Chinese Medicine (TCM) to remove blood stasis. Carbonized Typhae Pollen (CTP), a processed product of TP after being stir-fried, has been widely applied to clinical practice with its capability of hemostasis. However, the underlying mechanism of TP and CTP are still not fully elucidated and discrimination against TP and CTP remains a challenge. Aim of studyThe aim of this study is to investigate whether TP could remove blood stasis by promoting angiogenesis and the process of carbonizing it could enhance hemostatic effect. Meanwhile, some chemical markers for quality control of CTP had better to be found. Material and methodsThe changes of constituents between TP and CTP were analyzed by UPLC-QTOF-MS/MS. We investigated pro-angiogenic and hemostatic effects of TP and CTP in two zebrafish models: VRI-induced ISV insufficiency model and Ator-induced cerebral hemorrhage model. Subsequently, quantitative real-time PCR (qRT-PCR) was applied to investigate the mechanism of pharmacological effects. Finally, chemometric method was applied to find chemical markers. ResultsA total of 19 compounds were identified in qualitative analysis. The loss rate of each compound was calculated and compared. Two compounds (huaicarbon A/B) could only be detected in CTP and the content of flavonoid glycosides in CTP was significantly decreased compared with TP. The average content of the three identified flavonoid aglycones (quercetin, isorhamnetin and kaempferol) was increased about 30 percent in CTP. TP promoted pro-angiogenesis by up-regulating the expression of VEGFA, flt1 and kdr. After heating process, the pro-angiogenic activity was reduced and hemostatic activity was enhanced in CTP. Then qRT-PCR analysis found that CTP could significantly up-regulate the expression of VEGFA and vWF. In the discovery of markers, 6 chemical markers for discrimination of TP and CTP were obtained by chemometric method. ConclusionOur research indicated that the pro-angiogenic activity of TP was involved in VEGF signaling pathway. After processing, hemostatic activity of CTP has been enhanced by up-regulating the expression of VEGFA and vWF. A chemical marker database was established to provide a scientific evidence for quality control, mechanism and the clinical application of TP and CTP.

Shi Xiao San ameliorates the development of adenomyosis in an ICR mouse model

The use of Shi Xiao San (SXS), composed of Pollen Typhae Angustifoliae and Faeces Trogopterori, can be traced back to the Song dynasty. Traditionally, SXS has been used to treat irregular menstruation, pelvic pain, progressive dysmenorrhea, and postpartum lochiorrhea. The management of adenomyosis (AM) is challenging and to the best of our knowledge there are currently no effective therapeutic strategies. Therefore, the aim of the present study was to investigate the effect of SXS on the development of adenomyosis in a mouse model. AM was induced in 60 neonatal female ICR mice by administering tamoxifen; 10 randomly selected mice were used for model identification via histopathological examination and 10 mice treated with the solvent alone were used as the normal controls. A total of sixty days after birth, the mice treated with AM were randomly divided into four groups and administered one of the following treatments: Low-dose SXS (55 mg/kg); high-dose SXS (110 mg/kg); danazol (1 mg/20 g body weight); or no treatment (model group); at the same time, the normal control group received no treatment. After 2 months of treatment, hotplate and tail-flick tests were used to assess the response to noxious thermal stimuli in the mice, and plasma samples were collected to measure corticosterone levels. Hematoxylin and eosin staining scores of myometrial infiltration and the number of AM nodules were evaluated. Furthermore, the expression of genes associated with AM-related pain was also analyzed. The results from the present study indicated that treatment with SXS decreased myometrial infiltration, alleviated generalized hyperalgesia, and lowered plasma corticosterone levels in mice with induced AM. These findings suggest that SXS effectively attenuated the development of AM, and may serve as a promising treatment approach for AM treatment.

Quality assessment of Typhae Pollen Carbonisata based on chromaticity analysis combined with UPLC fingerprinting and thrombin activity.

Typhae pollen (TP) has been used as an anticoagulant in traditional Chinese medicine and throughout Asia. Typhae Pollen Carbonisata (TPC), a processed product of TP, has hemostatic properties. TPC is produced by frying TP, and the degree of processing can be judged by the colour change from yellow to brown. To establish a novel method for quality assessment of TPC and discriminate TPC from underdone products and overdone products. The Commission Internationale de l'Eclairage (CIE) L* a* b* colour space values of TP and TPC were measured to establish the colour model of TPC. Ultra-performance liquid chromatography was developed for fingerprinting. Thrombin activity promoting rates, clotting time, and bleeding time illustrated the difference in the hemostatic effect of the processed products. Chemometric approaches were performed to reveal the correlation between components and colour values or thrombin activity. Reference ranges of colour values and mathematical functions of TPC were established. The developed method was found to be fast, economic, sensitive, and stable. Fingerprints and thrombin activity in conjunction with partial least squares (PLS) demonstrated that peaks 2, 4, 7, 30, and 36 (isorhamnetin) were the main contributors for colour values and hemostatic activity of TPC. TPC and its unqualified products can be effectively distinguished based on chromaticity analysis, which provides a powerful tool for the comprehensive evaluation of the quality of Chinese herbal medicines.

Typhae pollen polysaccharides protect hypoxia-induced PC12 cell injury via regulation of miR-34a/SIRT1.

This current research was performed to investigate the role of typhae pollen polysaccharides (TPP) in hypoxia-treated PC12 cell which was an in vitro cell model of cerebral ischemia. Hypoxia-treated cells were treated with TPP for 12 h. Cell viability and apoptosis were detected by 3-(4,5-dimethylthiazol-2-yl)-2 5-diphenyl-2H-tetrazolium bromide (MTT) assay and flow cytometry, respectively. Cell apoptotic proteins and PI3K/AKT and Ras/Raf/MEK/ERK signal pathway–associated proteins were also examined by western blot. Furthermore, abnormal expression of miR-34a and silent information regulator 1 (SIRT1) was achieved by transfection. Besides, the expression of miR-34a and SIRT1 was examined by quantitative real-time polymerase chain reaction (qRT-PCR). The expression of SIRT1 was detected by qRT-PCR and western blot. The relationship between miR-34a and SIRT1 was verified by luciferase assay. We found that TPP enhanced cell viability and inhibited apoptosis in hypoxia-treated PC12 cells. Moreover, TPP increased the accumulated levels of Bcl-2 while decreased expression of Bax, cleaved Caspase-3, and cleaved PARP. TPP downregulated miR-34a expression while induced by hypoxia. Further results showed that miR-34a overexpression reversed the results led by TPP in cell viability, apoptosis, and its related proteins. In addition, SIRT1 was upregulated by TPP and was verified to be a target of miR-34a. Silence of SIRT1 led to the opposite results led by TPP. In the end, TPP activated PI3K/AKT and Ras/Raf/MEK/ERK signal pathways. In conclusion, TPP plays important roles in regulating cell viability and apoptosis in hypoxia-treated PC12 cells via modulating miR-34a/SIRT1, as well as activating PI3K/AKT and Ras/Raf/MEK/ERK signal pathways.

Near-infrared spectroscopy to assess typhaneoside and isorhamnetin-3-O-glucoside in different processed products of pollen typhae

With the ever increasing importance of testing drug quality, rapid analytical methods are needed for supervision of Chinese herbal medicines. Near-infrared spectroscopy is one of the most powerful tools in quality assessment of Chinese herbal medicines. In this work, near-infrared spectroscopy was applied to develop a rapid method for quantitative determination of typhaneoside and isorhamnetin-3-O-glucoside in different processed products of Pollen Typhae. A total of 71 batches of samples were collected from different regions in China. After acquisition of near-infrared spectra, different pre-processing methods were compared, and a competitive adaptive reweighted sampling algorithm was used to perform the variable selection. Then a partial least squares regression algorithm was applied to build the quantitative models. The root mean square error of calibration, root mean square error of cross validation, and root mean square error of prediction were 0.0190, 0.0364, and 0.0158%, respectively, for a quantitative model of typhaneoside. The root mean square error of calibration, root mean square error of cross validation, and root mean square error of prediction were 0.0190, 0.0377, and 0.0170%, respectively, for a quantitative model of isorhamnetin-3-O-glucoside. Moreover, the relative prediction deviation values of both quantitative models were larger than 3, indicating good performance of the partial least squares (PLS) models. The results demonstrated that high accuracy prediction of typhaneoside and isorhamnetin-3-O-glucoside could be obtained by near-infrared spectroscopy, to allow an alternative method for quality assessment of different processed products of Pollen Typhae.

Typhaneoside prevents acute myeloid leukemia (AML) through suppressing proliferation and inducing ferroptosis associated with autophagy

Leukemia remains a fatal disease for most patients and effective therapeutic strategies are urgently required. Typhaneoside (TYP) is a major flavonoid in the extract of Pollen Typhae, showing significant biological and pharmacological effects. In the present study, we explored the effects of TYP on acute myeloid leukemia (AML) progression. The results indicated that TYP markedly reduced the cell viability of AML cells and arrested the cell cycle at the G2/M phase by regulating the expression of associated proteins. In addition, TYP significantly induced apoptosis in AML cells by promoting the activation of Caspase-3. Intracellular and mitochondrial reactive oxygen species (ROS) accumulation were highly detected in AML cells after treatment with TYP. Moreover, TYP clearly induced ferroptosis in AML cells, and this process was iron-dependent and attendant with mitochondrial dysfunction. We also found that TYP significantly triggered autophagy in AML cells by promoting the activation of AMP-activated protein kinase (AMPK) signaling, contributing to ferritin degradation, ROS accumulation and ferroptotic cell death ultimately. In conclusion, the findings above provided solid evidences that TYP could be a promising therapeutic agent to prevent AML progression by inducing apoptosis, ROS production, autophagy and ferroptosis.