Effect and mechanism of Panax notoginseng saponins on wound healing after anal fistula surgery in rats / 中国药房

OBJECTIVE To investigate the effect and mechanism of Panax notoginseng saponins （PNS） on wound healing after anal fistula surgery in rats by regulating the hypoxia-inducible factor-1α （HIF-1α）/vascular endothelial growth factor （VEGF）/ vascular endothelial growth factor receptor-2 （VEGFR2） signaling pathway. METHODS SD rats were selected to establish a postoperative rat model of anal fistula by infecting wound with Escherichia coli. The model rats were randomly grouped into model group， PNS low-dose and high-dose groups （15， 30 mg/cm2）， high-dose of PNS+2-methoxyestradiol （2ME2） group （PNS 30 mg/cm2+HIF-1α inhibitor 2ME2 4 mg/kg）， with 10 rats in each group. Another 10 normal rats were selected for back hair removal treatment as the control group. Each drug group was injected with the corresponding drug solution intramuscularly or （and） intraperitoneally， once a day， for 3 weeks. After the last administration， the wound healing rate （excluding the control group）， microvascular density （MVD）， the expression of collagen Ⅰ and fibronectin （FN） in the wound tissue were detected in each group； the levels of angiogenic factors [VEGF， E-mail：842710813@qq.com angiopoietin-Ⅰ （Ang-Ⅰ）， Ang-Ⅱ] in serum， the levels of inflammatory factors [interleukin-6 （IL-6） and IL-2] in serum binggui7183@163.com and wound tissue as well as the expressions of the related proteins of HIF-1α/VEGF/VEGFR2 signaling pathway in the wound tissue of rats were also detected in each group. RESULTS The MVD， the expression of collagen Ⅰ and FN in the wound tissue， and the levels of IL-6 and IL-2 in serum and wound tissue of rats increased significantly in the model group， compared to the control group （P＜0.05）， while the serum levels of VEGF， Ang- Ⅰ and Ang-Ⅱ decreased significantly （P＜0.05）. The wound healing rate， the MVD in wound tissue， the serum levels of VEGF， Ang-Ⅰ and Ang-Ⅱ， the expressions of collagen Ⅰ and FN in the wound tissue， and protein expressions of HIF-1α， VEGF and VEGFR2 in the PNS low-dose and high-dose groups increased significantly， compared to the model group （P＜0.05）， while the levels of IL-6 and IL-2 in serum and wound tissue decreased significantly （P＜0.05）； the high-dose PNS had a stronger effect （P＜ 0.05）. 2ME2 could weaken the effect of PNS on above indicators of rats after anal fistula surgery （P＜0.05）. CONCLUSIONS PNS can promote the production of angiogenic factors and inhibit the production of pro-inflammatory factors， thereby promoting wound healing in rats after anal fistula surgery. The above effects are related to the activation of HIF-1α/VEGF/VEGFR2 signaling pathway.

Panax notoginseng saponin relieving the inflammatory pain caused by complete Freund’s adjuvant by inhibiting the activation of astrocytes in mice / 解剖学报

Objective To analyse the analgesic effect and possible mechanism of panax notoginseng saponin (PNS) on mouse models of chronic inflammatory pain caused by complete Freund’s adjuvant (CFA). Methods A total of 48 male C57BL/ 6J mice were divided randomly into four groups: normal saline control group (Ctrl), CFA group (CFA), CFA + PNS group (CFA+PNS), CFA + dexamethasone (DEX) group (CFA+DEX). Von Frey filaments were used to detect mechanical pain in mice. Immunohistochemistry was used to detect the number and morphological changes of glial fibrillary acidic protein (GFAP) positive astrocytes. Western blotting was used to detect the expressions of GFAP, nucleotide-binding and oligomerization domain(NOD)-like receptor thermal protein domain associated protein 3 (NLRP3), apoptosis-associated speck-like protein containing a CARD (ASC), Caspase-1, interleukin (IL)-1β, and IL-18 in mice’s spinal cord segments in each group. Results Compared with the Ctrl group, mice in the CFA group showed a significant decrease in mechanical pain thresholds at day 1, day 3, day 5, day 7, and day 14. Additionally, there was a significant decrease in NLRP3, ASC, Caspase-1, IL-1β and IL-18 in the spinal cord of the mice. PNS intervention could relieve mechanical pain and down-regulate the expressions of NLRP3, ASC, Caspase-1, IL-1β and IL-18 in the spinal cord of mice, with no significant difference compared with the CFA+DEX group. CFA group mice had significantly more GFAP positive cells in their posterior horns than Ctrl group mice, as measured by immunohistochemistry; PNS intervention decreased the number of GFAP positive cells in the posterior horn of the spinal cord in model mice;DEX had no effect on the number of GFAP positive cells in the dorsal horn of spinal cord. According to Western blotting results, GFAP expression in the spinal cord of the CFA group was significantly more than that of the Ctrl group; PNS intervention significantly reduced GFAP expression in the spinal cord of CFA group mice;DEX had no effect on the expression of GFAP in the posterior horn of spinal cord. Conclusion PNS has a good alleviating effect on inflammatory pain, and its mechanism may be related to inhibition of astrocyte activation and NLRP3 inflammasome activation.

Mechanism of active components of "Notoginseng Radix et Rhizoma-Drynariae Rhizoma" in treatment of osteoporosis based on network pharmacology and in vitro cell experiment / 中国中药杂志

The present study aimed to explore the main active components and potential mechanisms of Panax notoginseng saponins(PNS) and osteopractic total flavone(OTF) in the treatment of osteoporosis(OP) through network pharmacology, molecular docking and in vitro cell experiments, which was expected to provide a theoretical basis for clinical applications. The blood-entering components of PNS and OTF were obtained from literature search and online database, and their potential targets were obtained from Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP) and SwissTargetPrediction. The OP targets were obtained by means of searching Online Mendelian Inheritance in Man(OMIM) and GeneCards. The common targets of the drug and disease were screened by Venn. Cytoscape was used to construct a "drug-component-target-disease" network, and the core components were screened according to the node degree. The protein-protein interaction(PPI) network of the common targets was constructed by STRING and Cytoscape, and the core targets were screened according to the node degree. GO and KEGG enrichment analysis of potential therapeutic targets were carried out by R language. Molecular docking was used to determine the binding activity of some active components to key targets by AutoDock Vina. Finally, HIF-1 signaling pathway was selected for in vitro experimental verification according to the results of KEGG pathway analysis. Network pharmacology showed that there were 45 active components such as leachianone A, kurarinone, 20(R)-protopanaxatriol, 20(S)-protopanaxatriol, and kaempferol, and 103 therapeutic targets such as IL6, AKT1, TNF, VEGFA and MAPK3 involved. PI3K-AKT, HIF-1, TNF and other signaling pathways were enriched. Molecular docking revealed that the core components had good binding ability to the core targets. In vitro experiments found that PNS-OTF could up-regulate the mRNA expression levels of HIF-1α, VEGFA and Runx2, indicating that the mechanism of PNS-OTF in treating OP may be related to the activation of HIF-1 signaling pathway, and thus PNS-OTF played a role in promoting angiogenesis and osteogenic differentiation. In conclusion, this study predicted the core targets and pathways of PNS-OTF in treating OP based on network pharmacology and carried out in vitro experimental verification, which reflected the characteristics of multi-component, multi-target and multi-pathway synergy of PNS-OTF, and provided new ideas for the future clinical treatment of OP.

Targeted trace ingredients coupled with chemometric analysis for consistency evaluation of Panax notoginseng saponins injectable formulations / 中国天然药物

Evaluating the consistency of herb injectable formulations could improve their product quality and clinical safety, particularly concerning the composition and content levels of trace ingredients. Panax notoginseng Saponins Injection (PNSI), widely used in China for treating acute cardiovascular diseases, contains low-abundance (10%-25%) and trace saponins in addition to its five main constituents (notoginsenoside R1, ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1, and ginsenoside Rd). This study aimed to establish a robust analytical method and assess the variability in trace saponin levels within PNSI from different vendors and formulation types. To achieve this, a liquid chromatography-triple quadrupole mass spectrometry (LC-MS/MS) method employing multiple ions monitoring (MIM) was developed. A "post-column valve switching" strategy was implemented to eliminate highly abundant peaks (NR1, Rg1, and Re) at 26 min. A total of 51 saponins in PNSI were quantified or relatively quantified using 18 saponin standards, with digoxin as the internal standard. This study evaluated 119 batches of PNSI from seven vendors, revealing significant variability in trace saponin levels among different vendors and formulation types. These findings highlight the importance of consistent content in low-abundance and trace saponins to ensure product control and clinical safety. Standardization of these ingredients is crucial for maintaining the quality and effectiveness of PNSI in treating acute cardiovascular diseases.

Panax notoginseng saponins prevent colitis-associated colorectal cancer via inhibition IDO1 mediated immune regulation / 中国天然药物

Colorectal cancer (CRC) is the third most lethal cancer and leading cause of cancer mortality worldwide. A key driver of CRC development is colon inflammatory responses especially in patients with inflammatory bowl disease (IBD). It has been proved that Panax notoginseng saponins (PNS) have anti-inflammatory, anti-oxidant and anti-tumor effects. The chemopreventive and immunomodulatory functions of PNS on colitis-associated colorectal cancer (CAC) have not been evaluated.This present study was designed to study the potential protective effects of PNS on AOM/DSS-induced CAC mice to explore the possible mechanism of PNS against CAC. Our study showed that PNS significantly alleviated colitis severity and prevented the occurrence of CAC. Functional assays revealed that PNS relieved immunosuppression of Treg cells in the CAC microenvironment by inhibiting the expression of IDO1 mediated directly by signal transducer and activator of transcription 1 (STAT1) rather than phosphorylated STAT1. Ultimately, Rh1, one of the PNS metabolites, exhibited the best inhibitory effect on IDO1 enzyme activity. Our study showed that PNS exerted significant chemopreventive function and immunomodulatory properties on CAC. It could reduce macrophages accumulation and Treg cells differentiation to reshape the immune microenvironment of CAC. These findings provided a promising approach for CAC intervention.

Panax notoginseng saponins ameliorated LPS-induced acute lung injury in mice by inhibiting the activation of NF-κB / 药学学报

Acute lung injury (ALI) is a kind of lung disease mainly caused by excessive inflammatory reaction. At present, there is a lack of effective therapeutic drugs in clinic. The aim of this study was to investigate the improvement effect of Panax notoginseng saponins (PNS) on ALI and its potential mechanism. The model of wild-type C57BL/6J mice was established by intratracheal instillation of 50 μL 25 mg·mL-1 lipopolysaccharide (LPS). 24 h later, 200 and 400 mg·kg-1 PNS was given intragastric, respectively. 24 h after administration, the improvement effect of PNS on ALI mice was evaluated by lung function, wet-to-dry weight ratio (W/D), total protein, interleukin 6 (IL6) and tumor necrosis factor α (TNFα) concentration of bronchoalveolar lavage fluid (BALF), expression levels of IL6 and TNFα in lung tissues, pathological changes of lung tissues and expression of inflammatory cells in BALF. The protein expression levels of NF-κB and its upstream kinases in Raw264.7 cells and ALI mice lung tissues were further detected to evaluate the potential mechanism of PNS improving ALI mice. The experimental scheme was approved by the Animal Experiment Ethics Committee of Shanghai University of Traditional Chinese Medicine. It was found that 400 mg·kg-1 PNS could significantly improve the lung function of ALI mice, reduce the contents of W/D, BALF total protein, IL6 and TNFα, neutrophils expression in BALF and the infiltration of inflammatory cells in lung tissue. In Raw264.7 cells and ALI mice lung tissue, PNS significantly reduced the expression of NF-κB, reduced the protein expression and phosphorylation of NF-κB, promoted the expression of IκBα, and inhibited the inflammatory response. This study showed that PNS can improve ALI by inhibiting the activity of NF-κB, inhibiting the release of inflammatory factors and inflammatory cells infiltration, alleviating lung inflammation.

Panax notoginseng Saponins Alleviate TGF-β1-induced Renal Tubular Epithelial Cell Injury by Inhibiting Autophagy / 中国实验方剂学杂志

ObjectiveTo investigate the role and mechanism of Panax notoginseng saponins （PNS） in inhibiting transforming growth factor-β1 （TGF-β1）-induced renal tubular epithelial cell injury. MethodNRK-52E renal tubular epithelial cells were cultured and divided into control group， TGF-β1 group，TGF-β1+12.5 mg·L-1 PNS group，TGF-β1+25 mg·L-1 PNS group and TGF-β1+50 mg·L-1 PNS group. After 48 hours of PNS intervention， the cells and the supernatant were collected， and cell morphology was observed by inverted microscope. Western blot was used to detect the expression of epithelial-mesenchymal transition （EMT）-related proteins and autophagy-related proteins. Flow liquid chromatography for multiple protein quantification and flow cytometry were employed to determine the content of inflammatory factors and apoptosis rate， respectively. ResultCompared with the conditions in the control group， after TGF-β1 induction， the cells showed a spindle-shaped change and the expression of E-cadherin was down-regulated （P<0.05）， while the expression of α-smooth muscle actin （α-SMA） was up-regulated （P<0.05）. After PNS treatment， most of the cells tended to be normal and reversed the occurrence of EMT. In addition， compared with the conditions in the control group， the level of TNF-α was increased while that of IL-10 was decreased， with elevated apoptosis rate （P<0.05） in the TGF-β1 group. After PNS treatment， the level of TNF-α was lowered while that of IL-10 was boosted with the increase of the dose， with reduced apoptosis rate （P<0.05）. Moreover， after TGF-β1 induction， the expression of autophagy-related proteins Beclin 1 and LC3Ⅱ/Ⅰ in renal tubular epithelial cells were up-regulated， while PNS inhibited their expression（P<0.05，P<0.01）. ConclusionPNS had a protective effect on TGF-β1-induced renal tubular epithelial cells， and the mechanism might be that it reduced inflammation and apoptosis by inhibiting autophagy， thus alleviating TGF-β1-induced injury.

Preparation and intestinal absorption of Panax notoginseng saponins chitosan nanoparticles / 中国中药杂志

In this experiment, Panax notoginseng saponins chitosan nanoparticles(PNS-NPs) were prepared by self-assembly and their appearance, particle size, encapsulation efficiency, drug loading, polydispersity index(PDI), Zeta potential, and microstructure were characterized. The prepared PNS-NPs were intact in structure, with an average particle size of(209±0.258) nm, encapsulation efficiency of 42.34%±0.28%, a drug loading of 37.63%±0.85%, and a Zeta potential of(39.8±3.122) mV. The intestinal absorption of PNS-NPs in rats was further studied. The established HPLC method of PNS was employed to investigate the effects of pH, perfusion rate, and different drugs(PNS raw materials, Xuesaitong Capsules, and PNS-NPs). The absorption rate constant(K_a) and apparent permeability coefficient(P_(app)) in the duodenum, jejunum, ileum, and colon were calculated and analyzed. As illustrated by the results, the intestinal absorption of PNS-NPs was increased in the perfusion solution at pH 6.8(P<0.05), and perfusion rate had no significant effect on the K_a and P_(app) of PNS-NPs. The intestinal absorption of PNS-NPs was significantly different from that of PNS raw materials and Xuesaitong Capsules(P<0.05), and the intestinal absorption of PNS-NPs was significantly improved.

Prevention of Deep Vein Thrombosis by Panax Notoginseng Saponins Combined with Low-Molecular-Weight Heparin in Surgical Patients / 中国结合医学杂志

OBJECTIVE@#To evaluate the efficacy of deep vein thrombosis (DVT) prevention among real-world surgical inpatients who received panax notoginseng saponins (PNS) combined with low-molecular-weight heparin (LMWH).@*METHODS@#A prospective cohort study was conducted among surgical patients between January 2016 and November 2018 in Xuanwu Hospital, Capital Medical University, Beijing, China. Participants received LMWH alone or PNS combined with LMWH for preventing DVT. The primary outcome was incidence of lower extremity DVT, which was screened once a week. Participants in the LMWH group were given LMWH (enoxaparin) via hypodermic injection, 4000-8000 AxalU once daily. Participants in the exposure group received PNS (Xuesaitong oral tablets, 100 mg, 3 times daily) combined with LMWH given the same as LMWH group.@*RESULTS@#Of the 325 patients screened for the study, 281 participants were included in the final analysis. The cohort was divided into PNS + LMWH group and LMWH group with 134 and 147 participants, respectively. There was a significant difference of DVT incidence between two groups (P=0.01), with 21 (15.7%) incident DVT in the PNS + LMWH group, and 41 (27.9%) incident DVT in the LMWH group. Compared with participants without DVT, the participants diagnosed with DVT were older and had higher D-dimer level. The multivariate logistic regression model showed a significant lower risk of incident DVT among participants in the PNS + LMWH group compared with the LMWH group (odds ratio 0.46, 95% confidence interval, 0.25-0.86). There were no significant differences in thromboelaslography values (including R, K, Angle, and MA) and differences in severe bleeding between two groups. No symptomatic pulmonary embolism occurred during the study.@*CONCLUSION@#Combined application of PNS and LMWH can effectively reduce the incidence of DVT among surgical inpatients compared with LMWH monotherapy, without increased risk of bleeding.

Panax notoginseng saponins regulates hematopoietic function of bone marrow in aplastic anemia mice via TLR4/TLR2-NF-kB pathway / 中国药理学通报

Aim To explore the effects of Panax notog- inseng saponins( PNS) on hematopoietic functions anrl regulation on the TLR4/TLR2-NF-kB signaling path¬way in immune-mediated aplastic anemia ( AA ) C57 mice.Methods C57BL/6 mice were randomly divid¬ed into control group, total body irradiation group ( TBI) , model group, cyclosporine treatment group, PNS low-dose group, medium-dose group and high-dose group.The immune-mediated A A mice model was es¬tablished by total body irradiation with 5.0 Gy X-ray and mixed lymphocyte infusion.The body weight was measured, the spleen and thymus index was calculated , bone marrow pathology, the levels of peripheral blood triline cells,bone marrow nucleated cells( BMCs) and the levels of serum TNF-cx , 1L-2 , 1L-10 were detected, and the expression of CD1 lc and proteins related to the TLR4/TLR2-N F- k B pathway were detected 15 days later.Results Compared with control group, body weight, thymus index, the number of peripheral blood triline cells, BMCs and serum 1L-10 levels of the mice in model group significantly decreased ( P < 0.05 ) , while spleen index, the serum TNF-a, IL-2 levels and the protein expression of CD 11 c, TLR4, TLR2 , MvD88 , Akt and NF-kB in hone marrow significantly increased ( P <0.05).Compared with model group, after PNS treatment, hodv weight, thymus index, the number of peripheral blood triline cells, BMCs and serum IL-10 levels increased.Spleen index,serum TNF-cx,lL-2 lev¬els and the expression of CD11 c, TLR4, TLR2, NF-kB and Akt in bone marrow decreased, and the therapeutic effect was not dose-dependent.There was no signifi¬cant change in the expression of MvD88 and MAPK proteins.Conclusions PNS can improve AA bone marrow injury, regulate immune disoders and promote hematopoiesis, which may be related to the regulation of the number of DCs and the TLH4/TLH2 - Akt- NF-kB pathway in bone marrow.

Preventive Effect of Panax notoginseng Saponins Combined With Bacillus subtilis on Colorectal Cancer Based on Microbiome / 胃肠病学

Background: Dysregulation of intestinal flora is a key risk factor for colorectal cancer (CRC). In recent years, traditional Chinese medicine preparations and probiotics have been increasingly applied in the prevention of CRC. Aims: To investigate the preventive effect of Panax notoginseng saponins (PNS) combined with Bacillus subtilis on CRC. Methods: Thirty female BALB/c mice were randomly divided into normal control group (NC group), model group, PNS group, Bacillus subtilis group and PNS combined with Bacillus subtilis group (PaB group). CRC mice model was constructed by azoxymethane (AOM)/dextran sulfate sodium (DSS) method. During the experiment, the mice were weighed, and disease activity index (DAI) score was evaluated. The length of colorectum and tumor number were measured. Serum interleukin (IL) - 6 and IL - 10 contents were determined by ELISA. 16S rRNA sequencing was used to analyze the composition of intestinal flora. Results: Compared with model group, DAI score was significantly decreased (P<0.001), colorectal length was significantly increased (P<0.001), number of tumor was significantly decreased (P<0.001), tumor volume was significantly decreased (P<0.01), serum IL-6 content was significantly decreased (P<0.000 1), and serum IL-10 content was significantly increased in PaB group (P<0.000 1). The results of intestinal flora sequencing showed that Simpson index was significantly decreased in PaB group than in model group (P<0.05), Shannon index and Chao index were significantly increased (P<0.05), abundance of Bacteroidota was significantly increased (P<0.01), abundances of Firmicutes, Helicobacter and Oscillibacter were significantly decreased (P all <0.05), abundance of Lactococcus was significantly increased (P<0.05). Conclusions: The combination of PNS and Bacillus subtilis can effectively alleviate the occurrence of CRC caused by AOM/DSS, and its mechanism may be related to the improvement of composition of intestinal microbial community.

Microscopic morphology observation of fracture healing process promoted by panax notoginseng saponins / Observación microscópica de la morfología del proceso de curación de fracturas promovida por panax notoginseng saponins

SUMMARY: The aim of this study was to explore promoting effect of external applying Panax Notoginseng Saponins (PNS) on fractures. For this analysis 18 New Zealand male rabbits were divided into control group, splintage group and PNS group. All rabbits were performed left radius fractures and natural healing, splintage healing and splintage coated with PNS healing. 2 rabbits in each group were sacrificed on day 14, day 28 and day 42 after surgery, separately. Atomic force microscope scanning and nanoindentation tests were performed on the callus sections. The particle size and roughness in PNS group was both less than that in splintage group. The elastic modulus of callus in PNS group was consistent with normal bone tissue started from day 28 after surgery, two weeks earlier than that in splintage group. PNS could significantly reduce fracture healing time and increase strength of callus.

RESUMEN: El objetivo de este estudio fue evaluar el efecto de la aplicación externa de Panax Notoginseng Saponins (PNS) en fracturas óseas. Se usaron 18 conejos machos de raza Nueva Zelanda divididos en grupos control, entablillado y PNS. Se realizaron fracturas del radio izquierdo y cicatrización natural en todos los animales, además de la cicatrización con entablillado y entablillado recubierto con PNS. Se sacrificaron, posterior a la cirugía, dos conejos de cada grupo los día 14, 28 y 42. Se realizaron pruebas de escaneo con microscopio de fuerza atómica y nanoindentación en las secciones de callos. El tamaño de la partícula y la rugosidad en el grupo de PNS fue menor que en el grupo entablillado. El módulo elástico del callo en el grupo de PNS fue consistente con el tejido óseo normal iniciado el día 28 después de la cirugía, dos semanas antes que en el grupo de entablillado. El PNS podría redu- cir significativamente el tiempo de curación de la fractura y aumentar la fuerza del callo.

Overview of systematic reviews of Panax notoginseng saponins in treatment of acute cerebral infarction / 中国中药杂志

To overview the systematic reviews of Panax notoginseng saponins in the treatment of acute cerebral infarction. CNKI, CBM, Wanfang, VIP, PubMed, Cochrane Library and EMbase databases were retrieved to collect the systematic reviews of the efficacy of P. notoginseng saponins in the treatment of acute cerebral infarction. The retrieval time was from the time of database establishment to January 2021. After two researchers independently screened out the literature and extracted the data, AMSTAR-2 scale was used to evaluate the methodological quality of the included systematic reviews, GRADE system was used to grade the quality of evidences of the outcome indicators, and the efficacy evaluation was summarized. A total of 5 systematic reviews were included. AMSTAR-2 evaluation results showed that 3 items were relatively complete, while 4 items had a poor overall quality. P. notoginseng saponins combined with conventional Western medicine therapy was superior to single conventional therapy in the recovery of neurological function, enhancement of the total effective rate in clinic, and improvement of activities of daily living. GRADE evaluation results showed that the quality of evidence was from low quality to very low quality. In conclusion, in the treatment of acute cerebral infarction, P. notoginseng saponins can improve the clinical efficacy, with a good safety but a not high methodological quality and a low evidence quality. It is suggested that high-quality clinical studies shall be further carried out to provide evidence-based basis for the application of P. notoginseng saponins in the treatment of acute cerebral infarction.

Panax Notoginseng Saponin Attenuates Gastric Mucosal Epithelial Cell Injury Induced by Dual Antiplatelet Drugs through COX and PI3K/Akt/ VEGF-GSK-3β-RhoA Network Pathway / 中国结合医学杂志

OBJECTIVE@#To elucidate the underlying mechanism of Panax notoginseng saponin (PNS) on gastric epithelial cell injury and barrier dysfunction induced by dual antiplatelet (DA).@*METHODS@#Human gastric mucosal epithelial cell (GES-1) was cultured and divided into 4 groups: a control, a DA, a PNS+DA and a LY294002+PNS+DA group. GES-1 apoptosis was detected by flow cytometry, cell permeability were detected using Transwell, level of prostaglandins E2 (PGE2), 6-keto-prostaglandin F1α (6-keto-PGF1α) and vascular endothelial growth factor (VEGF) in supernatant were measured by enzyme linked immunosorbent assay (ELISA), expression of phosphatidylinositide 3-kinase (PI3K), phosphorylated-PI3K (p-PI3K), Akt, phosphorylated-Akt (p-Akt), cyclooxygenase-1 (COX-1), cyclooxygenase-2 (COX-2), glycogen synthase kinase-3β (GSK-3β) and Ras homolog gene family member A (RhoA) were measured by Western-blot.@*RESULTS@#DA induced apoptosis and hyper-permeability in GES-1, reduced supernatant level of PGE2, 6-keto-PGF1α and VEGF (P<0.05). Addition of PNS reduced the apoptosis of GES-1 caused by DA, restored the concentration of PGE2, 6-keto-PGF1α and VEGF (P<0.05). In addition, PNS attenuated the alteration of COX-1 and COX-2 expression induced by DA, up-regulated p-PI3K/p-Akt, down-regulated RhoA and GSK-3β. LY294002 mitigated the effects of PNS on cell apoptosis, cell permeability, VEGF concentration, and expression of RhoA and GSK-3β significantly.@*CONCLUSIONS@#PNS attenuates the suppression on COX/PG pathway from DA, alleviates DA-induced GES-1 apoptosis and barrier dysfunction through PI3K/Akt/ VEGF-GSK-3β-RhoA network pathway.

Effect of Panax Notoginseng Saponins on Breast Cancer Cell Line 4T1 in Tumor-bearing Mice Through MEKK1/SEK1/JNK1/AP-1 Pathway / 中国实验方剂学杂志

Objective:To explore the potential mechanisms of Panax Notoginseng Saponins （PNS） on growth inhibition of breast cancer cell line 4T1 in tumor-bearing mice by investigating the mitogen-activated protein kinase kinase kinase 1 （MEKK1）/stress activated protein kinase （SAPK）/extracellular regulated protein kinases （Erk） Kinase （SEK1）/c-Jun N-terminal kinase 1 （JNK1）/activator protein-1 （AP-1） signaling pathways. Method:The 4T1 breast cancer mice model was established. Forty-eight mice with successful modeled and randomly divided into the low， medium and high-dose PNS groups （10， 20， 40 mg·kg-1） and the model control group （12 mice in each group）. The PNS groups received intraperitoneal injection with dosage of 10 mL·kg-1， while the controlled group was given the same dosage of saline. After administration with PNS for 28 days， tumor tissues were isolated， weighed， sliced and homogenized. Tumor cell apoptosis was detected by TdT mediated-dUTP nick end labeling （TUNEL） staining. The mRNA expressions of MEKK1， SEK1， JNK1 and AP-1 in tumor tissue were detected by Real-time polymerase chain reaction（Real-time PCR）. The protein expressions of MEKK1， SEK1， JNK1 and AP-1 in tumor tissue were detected by immunofluorescence staining and Western blot. Result:Compared with model group， the tumor weights of medium-dose and high-dose PNS groups were decreased significantly （P<0.05）. TUNEL staining showed that the number of apoptotic tumor cells increased with the rise of dosage of PNS （P<0.05）. The medium-dose and high-dose PNS groups showed a significant increase in the mRNA expressions of MEKK1， SEK1， JNK1 and AP-1 as well as the protein expressions of MEKK1， SEK1， JNK1 and AP-1 in tumor tissues （P<0.05）， with statistically significant differences （P<0.05）. Conclusion:PNS could inhibit the tumor growth of breast cancer cell line 4T1 in tumor-bearing mice， which may be related to the activation of MEKK1/SEK1/JNK1/AP-1 signaling pathways.

Exploratory study of APTT bioactivity detection assay for Xuesaitong injection / 中国药理学通报

Aim To explore the feasibility of APTT bioactivity detection for Xuesaitong injection. Methods The APTT values of rabbit plasma of samples/reference were detected by automatic coagulometer, and the data were analyzed by bioassay statistics. Results The results of repeated test of different rabbit plasma, multiple enterprises and various concentrations of Xuesaitong injection were stable. The APTT values of standard ginsenoside Rb1, ginsenoside Rg1, notoginsenoside R1 and Panax notoginseng saponins were compared with that of Xuesaitong injection, and the APTT values of the former three were not correlated with Xuesaitong injection; while the Panax notoginseng saponins were significantly correlated with Xuesaitong injection. The panax notoginseng saponins were used as the reference material, and the results were consistent with those of the physical and chemical analysis. Conclusions The active ingredients in Xuesaitong injection do not represent the effective ingredients, and its medicinal effect is the result of the combined action of various ingredients. Therefore, it is not suitable to simply look for some monomer as the control. This paper suggests using panax notoginseng saponins as the control substance to explore the biological activity study of injections for promoting blood circulation and removing blood stasis.

Improvement effect of panax notoginseng saponins on cardiac function in rats with chronic heart failure and its mechanism / 吉林大学学报(医学版)

Objective: To observe the improvement effect of panax notoginseng saponins on the cardiac function of the rats with chronic heart failure (CHF), and to investigate the possible mechanism. Methods: Abdominal aortic constriction was used to establish the CHF rat models. Sixty model rats were randomly divided into model group, positive control group, low, medium and high doses of panax notoginseng saponins groups ( n= 12). Another 12 rats were taken as sham operation group. Color floppier ultrasound was used to detect the left ventricular end-diastolic diameter (LVEDD)» left ventricular end-systolic diameter (LVESD), left ventricular posterior wall diastolic thickness (LVPWD), left ventricular ejection fraction (LVEF), cardiac output (CO) , left ventricular end-diastolic pressure (LVEDP), left ventricular systolic pressure (LVSP), maximal rate of increase of left ventricular pressure (+dp/dt max) and maximal rate of decrease of left ventricular pressure ( dp/dt max) of the rats in various groups∗ HE staining was used to observe the pathomorphology of myocardium tissue of the rats in various groups∗ TUNEL method was used to detect the apoptotic rates of cadiomyocytes of the rats in various groups∗ and Western blotting method was used to detect the expression levels of extracellular signal-regulated kinase (ERK), phosphorylation ERK (p-ERK), c-Jun NH2-terminal protein kinase (JNK), phosphorylation JNK (p-JNK), p38, and p-p38 proteins in myocardium tissue of the rats in various groups. ELISA was used to determine the serum levels of tumor necrosis factor-a (TNF-a) and interleukin-6 (IL-6) of the rats in various groups. Results: Compared with sham operation group, the LVEDI), LVESD, LVPWD, LVEDP and dp/dt max of the rats in model group were significantly increased (P

Neuroprotective effect of borneol combined with astragaloside IV and Panax notoginseng saponins in cerebral ischemia reperfusion injury rat model through Notch signaling pathway / 中草药

Objective: To observe the neuroprotective effect of borneol combined with astragaloside IV (AST IV) and Panax notoginseng saponins (PNS) on cerebral ischemia reperfusion injury (CIRI) rat model through Notch signaling pathway. Methods: SD rats were randomly divided into sham group, model group, borneol (7.5 mg/kg) group, AST IV (25 mg/kg) group, PNS (10 mg/kg) group, AST IV (10 mg/kg) + PNS (25 mg/kg) group, borneol (7.5 mg/kg) + AST IV (25 mg/kg) + PNS (10 mg/kg) low dose group, borneol (15 mg/kg) + AST IV (20 mg/kg) + PNS (50 mg/kg) high dose group and edaravone (4 mg/kg) group. Rats in sham group and model group were ig 0.5% CMC-Na, edaravone group was ip drug, and the other groups were ig corresponding drugs, twice a day with an interval of 12 h. The right middle cerebral artery of rat was blocked by a suture method 2 h after last administration to establish a CIRI model. After 2 h of ischemia and 22 h of reperfusion, the eurological function scores were scored and pathological changes of ischemic cortex in brain tissues of rats were observed by HE staining. The expressions of neuron specific nuclear (NeuN) and endothelial barrier antigen (EBA) in ischemic cortex of brain tissue were detected by immunohistochemistry. The expressions of vascular endothelial growth factor (VEGF), Notch1, and intracellular domain of Notch (NICD) in ischemic cortex of brain tissue were detected by Western blotting. Results: The score of neural dysfunction and cell damage rate in model group were significantly increased (P < 0.01); The score of nerve function defect and rate of cell damage in each administration group were significantly reduced (P < 0.05, 0.01), the effect of borneol + AST IV + PNS group was better than that of single drug and AST IV + PNS group (P < 0.05, 0.01). NeuN and EBA protein expressions were significantly decreased in the ischemic cortex of model group (P < 0.01), while NeuN and EBA protein expressions were significantly enhanced in each administration group (P < 0.05, 0.01), and the effect of borneol + AST IV + PNS group was better than that of single drug and AST IV + PNS group (P < 0.05, 0.01). In model group, VEGF protein expression was increased significantly (P < 0.05), while NICD and Notch1 protein expression had no significant change. The expression of VEGF, NICD and Notch1 protein were significantly up-regulated in borneol + AST IV + PNS group (P < 0.01), and the effect of combination of three drugs was better than that of single drug and AST IV + PNS (P < 0.05, 0.01). Conclusion: Borneol, AST IV, and PNS have the effects of preventing neuronal and cerebral microvascular damage after CIRI, and the effect of combination of three drugs was better than that of single drug and AST IV + PNS, which may be related to the activation of the Notch signaling pathway and up-regulation of VEGF expression, thereby, exerting protective effects on ischemic brain tissue.

Study on chemical constituents in acid hydrolysates of Panax notoginseng saponins / 中草药

Objective: To study the chemical constituents in acid hydrolysates of Panax notoginseng saponins (PNS). Methods: These compounds were separated and purified by column chromatography, and their structures were elucidated based on spectroscopic analyses (HR-ESI-MS, ESI-MS, 1H-NMR, 13C-NMR, HSQC and HMBC). Results: Eighteen compounds were obtained from the acid hydrolysates of PNS and characterized as dammar-25-ene-24-hydroperoxyl-3β,6α,12β,20S-tetraol (1), 6α,12β,20S-trihydroxy- dammarane-24-ene-3-O-β-D-glucopyranosyl-(1→2)-β-D-glucopyranoside (2), 6α,12β,20R-trihydroxy-dammarane-24-ene-3-O-β-D- glucopyranosyl-(1→2)-β-D-glucopyranoside (3), vina-ginsenoside-R8 (4), 24(S)-pseudo-ginsenoside-GQ (5), ginsenoside Rg5 (6), 20 (R)-ginsenoside Rg3 (7), 20(R)-ginsenoside Rk2 (8), 3β,12β-dihydroxy-dammar-(E)-20(22),24-diene-6-O-β-D-xylopyranosyl- (1→2)-β-D-glucopyranoside (9), 20(S)-ginsenoside Rg2 (10), ginsenoside SL1 (11), 20(R)-ginsenoside Rh1 (12), 20(22) E-ginsenoside Rh4 (13), 25-hydroxy-20(R) ginsenoside-Rh1 (14),3β,6α,12β,20(S)-20,25-epoxy-3,12-dihydroxy-dammarane-6-O-β-D-glucopyranoside (15), 20(R)-protopanaxadiol (16), 20(R)-protopanaxatriol (17), and 20(S)-protopanaxatriol (18). Conclusion: Compound 1 is a new triterpen saponin, and compounds 2-5 are isolated from P. notoginseng and acid dydrolysates of PNS for the first time.

Panax notoginseng saponins prevent colitis-associated colorectal cancer development: the role of gut microbiota / 中国天然药物

Gut microbiota dysbiosis is a risk factor for colorectal cancer (CRC) in inflammatory bowel disease (IBD). In this study, the effects of Panax notoginseng saponins (PNS) on colitis-associated CRC progression were evaluated on an azoxymethane (AOM)/dextran sulfate sodium (DSS) mouse model. In vivo, PNS significantly relieved AOM/DSS-induced colon tumorigenesis and development by reducing the disease activity index (DAI) scores and colon tumor load. The 16S rRNA data of fecal samples showed that the microbiome community was obviously destructed, while PNS could recover the richness and diversity of gut microbiota. Especially, PNS could increase the abundance of Akkermansia spp. which was significantly decreased in model group and negatively correlated with the progression of CRC. Moreover, ginsenoside compound K (GC-K) was evaluated on the effects of human CRC cells, which was the main bio-transformed metabolite of PNS by gut microbiota. Our data showed that PNS played important role in the prevention of the progression of CRC, due to their regulation on the microbiome balance and microbial bio-converted product with anti-CRC activity.