Codonopsis Pilosula Polysaccharide Research Articles

Codonopsis pilosula polysaccharide suppresses the progression of non-small cell lung cancer by triggering NLRP3/GSDMD-dependent pyroptosis

BackgroundNon-small cell lung cancer (NSCLC) represents a prevalent and challenging malignancy, often posing difficulties in treatment due to late-stage diagnosis and limited therapeutic options.ObjectiveThis study aims to elucidate the impact of Codonopsis pilosula polysaccharide (CPP) on NSCLC, exploring its potential anticancer molecular mechanisms.Materials and methodsPurchase A549 cells and treat them with varying concentrations of CPP, dividing them into groups. Use CCK-8 assays for cell proliferation and TUNEL assays for apoptosis detection. Perform Western blot (WB) to detect NF-κB p65, pNF-κB p65, and pyroptosis-related proteins. Measure ROS, inflammatory factors, and LDH levels using kits. Observe pyroptosis morphology under a microscope. Inoculate A549 cells subcutaneously into nude mice to create a xenograft model and treat them with daily CPP injections. Collect tumor tissues, measure tumor volume and mass. Perform TUNEL immunohistochemistry for apoptosis and H&E staining for tumor histology. Detect pyroptosis-related proteins with Western blot and immunohistochemistry, and assess inflammatory factors using ELISA and immunohistochemistry.ResultsThe research reveals a concentration-dependent inhibitory effect of CPP on A549 cell viability, with optimal efficacy observed at 40 μmol/L CPP. CPP demonstrates the capacity to induce apoptosis, mediate NF-κB activation, and accumulate reactive oxygen species (ROS). Notably, CPP promotes pyroptosis in A549 cells and in vivo, upregulating pyroptosis-related proteins. In vivo administration of CPP significantly inhibits tumor growth, accompanied by notable morphological and histological alterations. Additionally, both in vitro and in vivo studies confirm elevated levels of IL-1β and IL-18 in tumor tissues following CPP treatment.Discussion and conclusionIn conclusion, CPP, as a multifaceted agent, suppresses the progression of NSCLC by inducing NLRP3/GSDMD-dependent pyroptosis. This research provides valuable insights into the therapeutic potential of CPP and stimulates further exploration for its integration into future NSCLC treatments.

Neuroprotective Effect of Codonopsis pilosula Polysaccharide on Aβ25-35-Induced Damage in PC12 Cells via the p38MAPK Signaling Pathways.

Plant polysaccharides have attracted increasing attention due to their high efficiency and low toxicity. Codonopsis pilosula polysaccharide (CPP) is an essential substance extracted from Codonopsis pilosula, known for its excellent antioxidant and neuroprotective effects. However, it is still unclear how CPP improves nerve protection and what its underlying molecular mechanisms are. This study aimed to investigate the neuroprotective effect of CPP on Aβ25-35-induced damage in PC12 cells and its underlying molecular mechanisms. The neuroprotective effect of CPP was evaluated using Aβ25-35-induced damage in pheochFfromocytoma (PC12) cells as an in vitro cell model. The cells were treated with CPP alone or in combination with SB203580 (an inhibitor of p38MAPK) in Aβ25-35 culture. The cell viability was assessed using a 3-(4,5-Dimethylthiazol-2-yl)-2,diphenyltetrazolium (MTT) assay. Furthermore, reactive oxygen species (ROS) were detected using flow cytometry. The production levels of intracellular superoxide dismutase (SOD), dismutase (SOD), glutathione (GSH), catalase (CAT), and malondialdehyFde (MDA) were determined using the colorimetric method. Annexin V-FITC and propidium iodide (PI) staining, as well as 33258 were performed using fluorescence microscopy. Moreover, the effect of adding SB203580 was studied to determine the changes in cell apoptosis induced by CPP treatment and Aβ25-35 induction. The CPP markedly inhibited Aβ25-35-induced reduction in the viability and apoptosis of PC12 cells. CPP also reduced the Aβ25-35-induced increase in the expression of the apoptosis factors and the levels of free radicals (ROS and MDA) and reversed the Aβ25-35-induced suppression of antioxidant activity. Additionally, inhibition of p38MAPK via the addition of their antagonists reversed the observed anti-apoptosis effects of CPP. CPP can efficiently provide neuroprotection against Aβ25-35-induced damage in PC12 cells brought about via oxidation and apoptosis reactions, and the underlying mechanisms involve the p38MAPK pathways. Therefore, CPP could potentially be useful as a neuroprotective agent in natural medicine, pharmacy, and the food industry.

MiR-221 on protective oxidative induced by selenium modified Codonopsis pilosula polysaccharide

Oxidative stress plays an important role in various diseases. miR-221 has been reported to regulate oxidative stress. However, the mechanism of miR-221 in regulating oxidative stress induced by sCPPS5 remains unclear. This study aimed to investigate the protective effects and mechanisms of miR-221 on oxidative stress induced by sCPPS5. The expression of SOD, CAT, MDA, LDH, MMP, caspase-3 activity and apoptosis were measured. In addition, the key signaling factors in the Keap1-Nrf2-ARE signaling pathway were determined by real-time PCR and Western blot. Mice were employed to evaluate the effects of sCPPS5 and the possible mechanism in vivo. sCPPS5 promoted the expression of SOD and CAT and activated Keap1-Nrf2-ARE signaling pathway inhibit the MDA content, MMP, caspase-3 activity, apoptosis and LDH release rate after transfection with miR-221 mimics and inhibitors. Consistently, sCPPS5 has the potential to enhance the expression of antioxidant enzymes as well as upregulate mRNA expression of crucial signal proteins in vivo. miR-221 on oxidative stress protection induced by sCPPS5 possibly through regulating the Keap1-Nrf2-ARE signaling pathway in macrophages.

Study on the mechanism of Codonopsis pilosula polysaccharide inhibiting gastric cancer precancerous lesions by regulating Wnt/β-catenin signaling pathway

AimTo investigate the effect of Codonopsis pilosula polysaccharide (CPP) on precancerous lesions of gastric cancer (PLGC) and its mechanism. MethodsThe CPP with more than 90 % sugar content was prepared by water extraction and alcohol precipitation combined with column chromatography. MTT assay was used to analyze the effect of CPP on the survival rate of human normal gastric epithelial cells (GES-1) and the proliferation of human gastric adenocarcinoma cells (AGS). The PLGC animal model was induced by chemical combined with irregular diet. The alleviate effect and related mechanism of CPP at high and low doses (110 mg/kg and 440 mg/kg) on PLGC were further analyzed. ResultsCPP can significantly promote the proliferation of hypoxia-induced GES-1 cells and after 48 h of CPP treatment, the expressions of Wnt-1, β-catenin and TCF-4 in GES-1 cells were significantly increased (P < 0.05). CPP can significantly inhibited the growth of AGS cells and after 48 h of CPP treatment, the expression of Wnt-1, β-catenin and TCF-4 in AGS cells was significantly decreased (P < 0.05). In the PLGC animal model, CPP could alleviate the damage degree of PLGC, inhibit the weight loss of rats, and significantly improve the levels of serum markers. Further, Western blot results showed that in the gastric tissue of rats, CPP could significantly increase the expression of Wnt-1β-catenin and TCF-4, and significantly decreased the ratio of Bcl-2/Bax, and significantly increased the expression level of caspase-3. The results of serum metabolomics analysis of rats showed that CPP may improve PLGC by affecting glycine metabolism, serine metabolism, threonine metabolism and other pathways. ConclusionsThis study confirms that CPP can alleviate PLGC. At the cellular level, CPP can play a protective role in gastric mucosa by activating Wnt/β-catenin signaling pathway under inflammatory or oxidative stress conditions, and can promote the apoptosis of abnormal cells of gastric glands by inhibiting Wnt/β-catenin signaling pathway. At the animal level, CPP can not only inhibit PLGC by inhibiting Wnt/β-catenin signaling pathway, but also alleviate PLGC by promoting abnormal apoptosis in gastric tissue.

Effect of Codonopsis pilosula polysaccharide on the quality of sheep semen preservation at 4°C.

This study aimed to investigate the effect of Codonopsis pilosula polysaccharide (CPP) on the motility, mitochondrial integrity, acrosome integrity rate, and antioxidant ability of sheep sperm after preservation at 4°C. Semen from healthy adult rams were collected and divided into four groups with separate addition of 0, 200, 400, and 1,000 mg/L CPP. Sperm motility was analyzed using the Computer-Assisted Semen Analysis software after preservation at 4°C for 24, 72, 120, and 168 h. Sperm acrosome integrity rate was analyzed by Giemsa staining at 24, 72, and 120 h, and mitochondrial membrane integrity was analyzed by Mito-Tracker Red CMXRos. The total antioxidant capacity (T-AOC) and malondialdehyde (MDA) content of spermatozoa were measured after 120 h of preservation. The sperm viability and forward-moving sperm under 200 mg/L CPP were significantly higher than that in the control group at 72 h (61.28%±3.89% vs 52.83%± 0.70%, 51.53%±4.06% vs 42.84%±1.14%), and 168 h (47.21%±0.85% vs 41.43%±0.37%, 38.68%±0.87% vs 31.68%±0.89%). The percentage of fast-moving sperm (15.03%±1.10% vs 11.39%±1.03%) and slow-moving sperm (23.63%±0.76% vs 20.29%±1.11%) in the 200 mg/L group was significantly higher than control group at 168 h. The mitochondrial membrane integrity of the sperm in the group with 200 mg/L CPP was significantly higher than those in the control group after storage at 4°C for 120 h (74.76%±2.54% vs 65.67% ±4.51%, p<0.05). The acrosome integrity rate in the group with 200 mg/L (87.66%±1.26%) and 400 mg/L (84.00%±2.95%) was significantly higher than those in the control group (80.65%±0.16%) after storage for 24 h (p<0.05). CPP also increased T-AOC and decreased the MDA concentration after preservation at 4°C (p<0.05). Adding CPP could improve the T-AOC of sperm, inhibit lipid peroxidation, and facilitate semen preservation.

Codonopsis pilosula polysaccharide alleviates rotenone-induced murine brain organoids death through downregulation of gene body DNA methylation modification in the ZIC4/PGM5/CAMTA1 axis

Here, the protective mechanism of Codonopsis pilosula polysaccharide (CpP) against mouse brain organoids (mBO) damage was analyzed, and the rotenone affected the genomic epigenetic modifications and physiological activity of mouse brain organoids was examined. Pathological experiments have shown that rotenone significantly damaged the subcellular organelles of mouse brain organoids. According to RRBS-Seq, rotenone significantly promoted gene body hypermethylation modifications in mouse brain organoids. Molecular biology experiments have confirmed that rotenone significantly promoted the hypermethylation modification of Zic4, Pgm5, and Camta1 gene bodies in mouse brain organoids, and their expression levels were significantly lower than those of the control group. Bioinformatic analysis suggested that multiple binding motif of transcription factors ZIC4 (Zinc finger protein of the cerebellum 4) were present at the promoters of both the Pgm5 (Phosphoglucomutase 5) and Camta1 (Calmodulin binding transcription activator 1) genes. When the expression of Zic4 was silenced, the proliferation of mouse brain organoids was significantly reduced and the expression level of PGM5 was also significantly decreased. In addition, Codonopsis pilosula polysaccharide treatment of mouse brain organoids significantly reduced the cytotoxicity of rotenone, promoted cell cycle progression, increased intracellular glutathione activity, significantly induced the demethylation modification of the Zic4, Pgm5, and Camta1 gene bodies, and promoted the high expression of ZIC4 and PGM5. Therefore, the study confirmed that Codonopsis pilosula polysaccharide alleviated rotenone-induced mouse brain organoids death by downregulating DNA gene bodies methylation modification of the Zic4/Pgm5/Camta1 axis.

Injectable hydroethanolic physical gels based on Codonopsis pilosula polysaccharide for sustained anticancer drug delivery

The development of biocompatible carriers based on hydroethanolic physical gels for effectively encapsulating and delivering hydrophobic drug molecules is of particular interest. In this paper, we reported a novel hydroethanolic physical gel based on Codonopsis pilosula polysaccharide (CPP) prepared from the roots of C. pilosula. The gelation behaviors of the graded CPP fractions in a water-ethanol solvent system were evaluated, and the physicochemical and mechanical properties of the CPP-based gel (CPP-G) were characterized. The results indicated that CPP-G had consisted of a random physically crosslinked network formed by hydrophobic association of CPP chains and exhibited good mechanical strength, higher shear-thinning sensitivity and rapid, highly efficient self-recovering characteristics, ensuring superior performance in constructing injectable and self-recovering drug-loaded gels. Hydrophobic paclitaxel (PTX) and hydrophilic doxorubicin (DOX) were used as representative drugs to investigate the encapsulation and in vitro release behaviors of CPP-G, which exhibited long-term sustained release properties. Additionally, the evaluation of drug activity in drug-loaded gels further revealed the synergistic effect of CPP-G with the selected drugs on tumor inhibition against 4T1 and MCF-7 breast cancer cell lines. This work evaluated the feasibility of using the natural polysaccharide CPP to construct hydroethanolic physical gels and the applicability of the injectable drug-loaded gels for hydrophobic drug delivery.

Preparation, characterization and immune activity of Codonopsis pilosula polysaccharide loaded in chitosan-graphene oxide

The purpose of this study was to investigate the effects of chitosan graphene oxide Codonopsis pilosula polysaccharide (CS-GO-CPP) complex on the immune function of macrophage cells (RAW264.7). In this experiment, chitosan (CS) was combined with graphene oxide (GO) by electrostatic action to prepare CS-GO nanocomposites, and it was used as a carrier to load Codonopsis pilosula polysaccharide (CPP) onto CS-GO to prepare CS-GO-CPP. Using infrared spectroscopy detection, zeta potential detection, and thermogravimetric analysis, we conduct a preliminary analysis of the structure of CS-GO-CPP. Macrophages were employed to evaluate CS-GO-CPP immunomodulatory activity and the possible mechanism responsible for the activation of macrophages in vitro. The results showed that compared with CPP, CS-GO-CPP did not change the basic structure of polysaccharide, and its thermal stability was improved. 0.78– 12.5 μg·mL−1 of CS-GO-CPP could significantly promote the phagocytic activity of RAW264.7 cells (P < 0.05) and significantly increase NO content, IL-4 and IFN-γ secretion, the expression of CD40, CD86, and F4/80 (P < 0.05). CS-GO-CPP might activate the NF-κB signaling pathway and induce the nuclear translocation of NF-κB p65. In conclusion, CS-GO-CPP has a capacity to activate RAW264.7 cells for an improvement of immunomodulation activities, which might be through NF-κB signaling pathway.

Codonopsis pilosula Polysaccharide Improved Spleen Deficiency in Mice by Modulating Gut Microbiota and Energy Related Metabolisms.

Codonopsis Radix (CR) is an important traditional Chinese medicine used for the treatment of spleen deficiency syndrome (SDS). Codonopsis pilosula polysaccharides (CPP) in CR are considered to be responsible for tonifying the spleen function; however, the mechanisms of the polysaccharides have remained unclear. This study aimed to investigate the treatment mechanisms of CPP in SDS mice using a combinational strategy of 16S rRNA gene sequencing and targeted metabolomics. Here, studies demonstrated that CPP had invigorating effect in vivo in Sennae Folium-induced SDS in mice by organ indexes, D-xylose determination, gastrointestinal hormones levels and goblet cells observation. Antibiotic treatment revealed that the intestinal microbiota was required for the invigorating spleen effect of CPP. Furthermore, gut microbiota analysis found that CPP significantly enriched probiotic Lactobacillus and decreased the abundance of some opportunistic pathogens, such as Enterococcus and Shigella. The metabolic profile analysis of the colonic content revealed that 25 chemicals were altered significantly by CPP, including amino acids, organic acids, fatty acids, carbohydrates and carnitine etc., which are mainly related to “energy conversion” related processes such as amino acids metabolism, tricarboxylic acid cycle, and nitrogen metabolism. Spearman’s correlation assays displayed there were strong correlations among biochemical indicators-gut microbiota-metabolomics. In summary, these results provided a new perspective for CPP improving SDS by regulating energy metabolism related bacteria and pathways.

Preparation of Codonopsis pilosula polysaccharide microcapsules and its effect and mechanism on skin wound healing in rats.

In this study, after optimizing the extraction process of CPP (Codonopsis pilosula polysaccharides), CPPM (CPP microcapsules) were prepared. Subsequently, the structural characteristics and physicochemical properties were studied. The results showed that CPPM is a hollow sac-like structure with rough folds and protuberances and comes in spherical or ellipsoidal shapes with uniform particle size. CPPM has certain swelling degree, low hardness, good adhesion, and stability. Then, the effect of CPPM on wounds repair was investigated by a rat model. The results showed that CPPM could improve the wound healing rate. Histological evaluation showed CPPM could promote neovascularization and fibroblast proliferation. By investigating the healing mechanism, it was found that CPPM increased the hydroxyproline content in granulation tissue and had an excellent antioxidant ability, and then inhibited lipid peroxidation, in addition, it significantly increased the transcript levels of VEGF and miRNA-21 genes, indicating that CPPM play an influential role in vascular remodeling during wound healing by up-regulating the expression of VEGF and miRNA-21 genes.

Extraction, purification, and biological activities in vivo of a novel fructose-rich polysaccharide from Codonopsis pilosula

The Codonopsis pilosula polysaccharide have been widely extracted through water dissolution and ethanol precipitation, and presented immunoregulatory and antioxidant activities. However, the polysaccharide from the supernatant was always discarded and rarely reported. In the present study, the extraction process, preliminary structural characteristics, immunoregulatory and antioxidant activities in vivo of C. pilosula polysaccharide prepared from the supernatant after 80% ethanol precipitation were evaluated and analyzed. Results showed that the optimized conditions for fructose-rich polysaccharide (Fru/Glc ratio of 2.73:1.00 with average molecular weight of 3.24 × 103 Da) extraction were determined as follows: liquid to material ratio of 40 mL/g, extraction time of 60 min, ultrasonic power of 370 W, and extraction temperature of 60 °C, and the yields reached to 6.52 ± 0.36%. Animal experiments demonstrated that the polysaccharide could effectively protect the immune organs, enhance the immune cells activities, and improve the leukocytes amounts and the expression levels of immune-related cytokines dose-dependently in CTX-treated mice, which was mainly attributed to the enhancement of antioxidant enzymatic activities and suppression of oxidative damage. These results suggested that the polysaccharide had the potential to be a natural antioxidant supplement due to the feasibility of industrial production and practical application.

Codonopsis pilosula polysaccharide in synergy with dacarbazine inhibits mouse melanoma by repolarizing M2-like tumor-associated macrophages into M1-like tumor-associated macrophages

BackgroundThe incidence and associated mortality of melanoma have increased significantly in recent years but treatment options are plagued with many undesirable side effects. Traditional Chinese herbal medicine polysaccharides are gaining increasing attention due to their potential role in the treatment of chronic diseases including tumors and the regulation of the immune system. MethodsIn this study, the potential effects of Ganoderma lucidum crude polysaccharides (GLCP) and Codonopsis pilosula crude polysaccharides (CPCP) on melanoma in C57 mice were explored. In addition, the inhibition and repolarization effect of digested Codonopsis pilosula polysaccharide (dCPP) on the proliferation of tumor-associated macrophages (TAMs) with M2-like phenotype induced by IL-4 were investigated. ResultsThe results showed that the various polysaccharides could significantly reduce tumor volume in melanoma mice. GLCP and GLCP + CPCP could further significantly reduce the number of CD68+ macrophages in tumors and also prolong survival in melanoma mice to a certain extent. Significantly, dCPP could inhibit the proliferation of IL-4-induced M2-like TAMs, and significantly increase the mRNA expression levels of IL-1, IL-6, iNOS and TNF-a, thereby promoting the repolarization of M2-like TAMs to M1-like TAMs. ConclusionOverall, it could be deduced that GLCP, CPCP and dCPP hold great potential as safe therapeutic options for melanoma and an immune-modulator which may require further exploration.

Stimulation of Codonopsis pilosula Polysaccharide on Bifidobacterium of Human Gut Bacteria In Vitro.

Objective To evaluate the prebiotic effects of Codonopsis pilosula polysaccharide (CPP) on human gut bacteria in vitro. Methods Codonopsis Radix was extracted with water at 100°C, and the extract was precipitated by 80% ethanol to obtain CPP. Human fresh fecal samples were collected from three healthy adults and used to ferment CPP. The fermented samples were collected to be analyzed by 16S rRNA sequencing. Results The results showed that CPP exhibited significantly the stimulation on the growth of genus Bifidobacterium of human gut bacteria (Padj < 0.05). Although CPP also exhibited regulative trends on the genera including Acidaminococcus, Bilophila, Dorea, and Eggerthella, no significant differences were observed (Padj > 0.05), which was likely associated with the limited samples (n = 3). Conclusion CPP has the potential to stimulate the growth of Bifidobacterium of the human gut bacteria and to be benefit to human health.

Hot Water Extraction of Crude Polysaccharide from Codonopsis pilosula and Determination of the Rheological Properties

The extraction condition of crude polysaccharides from Codonopsis pilosula by hot water reflux method was studied and the results analyzed by kinetic model. Then, the extraction rate and maximum extraction temperature were determined. furthermore, the thermal and pH stability and the rheological properties of the crude polysaccharide were also studied. crude Codonopsis pilosula polysaccharide (CCPP) under different conditions was extracted by hot water reflux method, the content was determined by phenol-sulfuric acid and DNS method, and the kinetic model of the extraction process was established. The viscosity was determined under different solute concentration, salt concentration, rotation speed, pH, and temperature. The thermal stability and pH stability were determined; the protein content and sulfate ion content were determined by the coomassie brilliant blue method and barium sulfate turbidimetry method, respectively. The yield of CCPP by the hot water reflux method was 25.72%. At 50 oC, the maximum extraction concentration, the rate constant and the linear relationship between the rate constant and the time were obtained at different ratios of material to liquid. The extraction process of this experiment coincided with Fick`s second law, R2 was greater than 0.88 and with the increase of extraction time, the dissolution rate increased, but with the increase of the ratio of material to liquid, the dissolution rate was decreased. CCPP showed a loss of polysaccharide fluidity as temperature increased and was affected by pH change. The apparent viscosity was affected by a change in solute concentration, temperature, pH, rotation speed and salt and sucrose concentrations.

The feeding of dietary Codonopsis pilosula polysaccharide enhances the immune responses, the expression of immune-related genes and the growth performance of red swamp crayfish (Procambarus clarkii)

Polysaccharides have many functions in aquatic animals and are widely used as immunopotentiators. However, despite the emergence of serious diseases, few studies have explored the effects of Codonopsis pilosula polysaccharide (CPP) on crustaceans. We studied the effects of CPP on the growth performance, nonspecific immunity, antioxidant activity and disease resistance of red swamp crayfish (Procambarus clarkii). Healthy crayfish (5.80 ± 0.1 g) were fed diets supplemented with 0% (control), 0.05%, 0.1%, 0.15%, 0.20%, and 0.30% CPP for 8 weeks. At the end of the 8-week feeding trial, the optimal final body weight (FBW), weight gain (WG), specific growth rate (SGR), and feed conversion ratio (FCR) were observed in the crayfish fed the diets with 0.15% and 0.20% CPP, followed by those fed the diet with 0.30% CPP and then those fed the diet with 0.10% CPP, whereas the values of these parameters were obtained with the control crayfish (P < 0.05). The crayfish fed the diets with 0.15% and 0.20% CPP exhibited a significantly higher total hemocyte count (THC) and significantly increased phenoloxidase (PO), lysozyme (LZM), hemocyte (Hc), acid phosphatase (ACP) and alkaline phosphatase (AKP) compared with those belonging to the other groups (P < 0.05). The crayfish fed the diets with 0.15% and 0.2% CPP exhibited significantly higher total superoxide dismutase (T-SOD) and glutathione peroxidase (GPx) activities, a significantly increased total antioxidant capacity (T-AOC) and a significantly lower malondialdehyde (MDA) content compared with the other groups (P < 0.05), which indicated that antioxidant capacity was significantly induced by the CPP-supplemented diets. Significantly upregulated expression of immune-related genes (anti-lipopolysaccharide factors (alf), peroxiredoxin (prx5), cathepsin B (ctsb), mitochondrial manganese superoxide dismutase (mtMnsod), cyclophilin A (cypa), glutathione peroxidase (gpx), Toll-like receptor 3 (tlr3), and heat shock protein 70 (hsp70)) was detected in the crayfish fed the diets supplemented with 0.15% and 0.20% CPP diet compared with the levels observed in the control crayfish. These results showed that dietary CPP supplementation greatly improved the growth, immunity and antioxidant capacities of crayfish, and according to the observed results, 0.15%–0.2% is the recommended optimal level of CPP dietary supplementation for crayfish.

The structural characterization and immune modulation activitives comparison of Codonopsis pilosula polysaccharide (CPPS) and selenizing CPPS (sCPPS) on mouse in vitro and vivo

Codonopsis pilosula polysaccharide (CPPS) and selenizing CPPS (sCPPS) were prepared and identified by a combination of chemical and instrumental analysis. Their immune modulation activities were compared by lymphocyte proliferation and flowcytometry tests in vitro or serum antibody responses and cytokines with immunization against OVA mice in vivo. The results showed that the sCPPS was successfully modified in selenylation. In vitro, the sCPPS were more effective compared with CPPS in promoting lymphocyte proliferation synergistically with PHA or LPS and increasing the ratio of CD4+ to CD8 + T cells. In vivo, sCPPS could significantly raised IgG, IgM, IFN-γ, IL-2 and IL-4 contents in the serum of mouse against OVA in comparison with CPPS. These results indicate that selenylation modification can enhance the immune modulation activitives of CPPS. sCPPS would be as a component drug of new-type immunoenhancer.

The inhibitory effect of phosphorylated Codonopsis pilosula polysaccharide on autophagosomes formation contributes to the inhibition of duck hepatitis A virus replication

Duck hepatitis A virus type 1 (DHAV) infection causes duck viral hepatitis and results in enormous loss to poultry farming industry. We reported that phosphorylated Codonopsis pilosula polysaccharide (pCPPS) inhibited DHAV genome replication. Here we further explored its underlying antiviral mechanisms. Autophagosomes formation is essential for the genome replication of picornaviruses. In this study, Western blot, confocal microscopy observation, and ELISA methods were performed to analyze polysaccharides' effects on autophagy by the in vitro and in vivo experiments. Results obtained from in vitro and in vivo experiments showed that Codonopsis pilosula polysaccharide did not play a role in regulating autophagy and had no therapeutic effects on infected ducklings. However, pCPPS treatment downregulated LC3-II expression level activated by DHAV and rapamycin, indicating the inhibition of autophagosomes formation. The interdiction of autophagosomes formation resulted in the inhibition of DHAV genome replication. Further study showed that pCPPS treatment reduced the concentration of phosphatidylinositol-3-phosphate (PI3P), an important component of membrane, in cells and serum, and consequently, autophagosomes formation was downregulated. In vivo experiments also verified the therapeutic effect of pCPPS. Phosphorylated Codonopsis pilosula polysaccharide treatment increased the infected ducklings' survival rate and alleviated hepatic injury. Our studies verified the effects of pCPPS against DHAV infection in duck embryo hepatocytes and ducklings and confirmed that phosphorylated modification enhanced the bioactivities of polysaccharides. The results also stated pCPPS's antiviral mechanisms, provided fundamental basis for the development of new anti-DHAV agents.

Effect of Codonopsis pilosula Polysaccharides on the Growth and Motility of Hepatocellular Carcinoma HepG2 Cells by Regulating β-Catenin/TCF4 Pathway

Objective. To study the effect of Codonopsis pilosula polysaccharide (CPP) on the growth and motility of HepG2 cells and its possible mechanism. Methods. Cells were randomly divided into Control group, CPP (5 μM) group, CPP (10 μM) group, and CPP (20 μM) group. The proliferation, invasion, migration ability, and expression of proteins involved in the epithelial-mesenchymal transition (EMT) and signaling pathway of HepG2 cells were detected by CCK8 assay, BrdU staining, Transwell, Scratch test, and Western blot, respectively. Results. Codonopsis pilosula polysaccharide inhibited the proliferation of HepG2 cells cultured in vitro along with the expression level of Ki67 and PCNA protein (P&lt;0.05), decreased the number of invasive cells (P&lt;0.05), and reduced the scratch closure rate (P&lt;0.05). It also adjusted the expression of vascular endothelial growth factor (VEGF), E-cadherin, and N-cadherin (P&lt;0.05). Other than that, downregulation of β-catenin, TCF4, and c-Myc protein expression (P&lt;0.05) was observed as well. Conclusion. Codonopsis pilosula polysaccharide can inhibit the proliferation and motility of HepG2 cells cultured in vitro, and the underlying mechanism is proposed to be related to the inhibition of the β-catenin/TCF4 pathway.

In vitro and in vivo immunomodulatory effects of fucoidan compound agents

Fucoidan extracted from brown algae displays diverse biological activities. In the present study, fucoidan was mixed with Chinese herb extracts, and the in vitro and in vivo immunomodulatory effects of two fucoidan compound agents were evaluated. The results showed that fucoidan from Kjellmaniella crassifolia (KF) and Undaria pinnatifida (UF) were sulfated polysaccharides, Astragalus polysaccharide (AP) was composed of α-d-glucose, and Codonopsis pilosula polysaccharide (CPP) was a furanose. Furthermore, fucoidan compound agents stimulated mouse macrophage RAW264.7 cell proliferation and enhanced the secretion of granulocyte-macrophage colony stimulating factor (GM-CSF) and tumour necrosis factor-α (TNF-α) in vitro. In addition, KCA (KF + AP + CPP) and UCA (UF + AP + CPP) could improve the nonspecific immunity and the specific immunity of BALB/c mice. Fucoidan compound agents also increased the secretion of GM-CSF, TNF-α, interleukin (IL)-4 and IL-10 in vivo. Therefore, we confirmed that fucoidan compound agents have promise for development as supplementary immunopotentiators.

Codonopsis pilosula Polysaccharide Attenuates Tau Hyperphosphorylation and Cognitive Impairments in hTau Infected Mice.

Codonopsis pilosula polysaccharide (CPPs), a natural products with potentially lower toxicity and better bioavailability has been used in traditional Chinese medicine for 1000s of years and a neuroprotective polysaccharide mitigates tau pathology in Alzheimer’s disease (AD) mouse model. However, whether CPPs can relieve AD pathology and cognitive defects remains poorly understood. Here we reported that CPPs remarkably increased the cell viability and PP2A activity, decreased tau phosphorylation in HEK 293/tau cells. Next, we employed an adeno-associated virus serotype 2 (AAV2)-induced expression of human full length tau (hTau) in C57/BL6 mice to mimic AD tau pathology. One month intragastric administration of CPPs significantly increased PP2A activity and reduced tau phosphorylation at Ser199, Ser202/Thr205 (AT8) and Thr231 in hippocampus of AAV2-hTau infected mice. Furthermore, behavioral tests revealed that CPPs rescued hTau overexpression induced cognitive defects while CPPs significantly increased the fEPSP slope and synaptic proteins including synaptotagmin and synaptophysin. Together, our data suggest that CPPs might prevent AD-like tau hyperphosphorylation via activation of PP2A and attenuates AD-like cognitive impairments through restoring the synaptic plasticity and synaptogenesis. In conclusion, our findings suggest that CPPs might be a potential candidate compound for the treatment of tau related diseases.