Extraction method of polysaccharide of overground part of cichorium intybus, polysaccharide and related application and evaluation method

Abstract

The invention relates to the field of biological extraction, substance analysis, application and effect evaluation, in particular to an extraction method of polysaccharide of an overground part of chicory, the polysaccharide and a related application and evaluation method. The method comprises the following steps: providing an extraction method of the polysaccharide of the overground part of the chicory, carrying out reflux extraction, concentration, precipitation, centrifugation, washing and freeze-drying to obtain the polysaccharide, and carrying out molecular weight distribution and monosaccharide composition analysis on the polysaccharide, and the polysaccharide has antioxidant and anti-inflammatory activity, can be used for preparing products for protecting myocardial cells and the like, and has wide application prospects. The invention also discloses a composition containing the polysaccharide and an evaluation method for comparing the myocardial protection activity of the polysaccharide at different parts of the cichorium intybus. The application achieves the technical effects of efficiently extracting the polysaccharide of the overground part of the chicory, determining the characteristics and effects of the polysaccharide, providing a basis for development of products related to cardiovascular disease prevention and treatment, and providing a method for evaluating the myocardial protection activity of the polysaccharide of different parts of the chicory.

Description

Technical Field

[0001] This application relates to the fields of biological extraction, material analysis, application and effect evaluation, and in particular to a method for extracting polysaccharides from the aerial parts of chicory, the polysaccharides and related applications and evaluation methods. Background Technology

[0002] Chicory, a widely cultivated plant used both as food and medicine, is gradually gaining prominence in the fields of food science, nutrition, and pharmacology. It boasts a long history of traditional use, and in-depth modern pharmacological research has revealed its diverse biological activities, including antioxidant, anti-inflammatory, anti-tumor, hepatoprotective, and metabolic regulatory effects. Chicory roots are an important commercial source of inulin, which, as a prebiotic dietary fiber, plays a vital role in the food industry, promoting gut health, regulating lipid and glucose metabolism, and improving overall health. Chicory polysaccharides, key active components of chicory phytochemicals, exhibit excellent performance in scavenging free radicals, regulating immunity, and modulating oxidative stress-related signaling pathways. In recent years, with the deepening research on chicory polysaccharides, more and more biological activities have been revealed, opening up broader prospects for its application in the food and health-related industries, such as the development of functional foods and the production of nutritional supplements.

[0003] Traditional methods for extracting chicory polysaccharides have largely focused on the roots. Extraction of chicory polysaccharides or inulin from chicory roots has been extensively studied and optimized under various temperature, time, and solvent conditions. Common methods include hot water extraction at different temperatures, multiple extraction cycles within different time ranges, and the use of various solvents to improve extraction efficiency. Typically, chicory roots are ground into powder, then refluxed with an appropriate amount of distilled water for extraction, followed by a series of steps including concentration, precipitation, and washing to obtain the polysaccharide components. However, despite increasing evidence that the aerial parts of chicory are rich in polysaccharides and possess biological potential, they are often discarded as agricultural byproducts in food production. Current research on polysaccharides from different parts of chicory is mainly limited to preliminary comparisons of polysaccharide content in different plant parts, lacking a comprehensive evaluation of their structural characteristics or pharmacological properties. Most studies simply compare the differences in polysaccharide content between roots and aerial parts without in-depth exploration of their structural features and specific pharmacological manifestations.

[0004] Currently, systematic comparative studies on polysaccharides derived from chicory roots and aerial parts are still lacking, hindering a comprehensive understanding of chicory polysaccharides and their wider application in the food and nutritional supplement industry. Because it is unclear whether the structural characteristics of aerial polysaccharides are comparable to those in the roots, and whether they exhibit similar levels of bioactivity, this potentially polysaccharide-rich and bioactive resource of chicory aerial parts is being wasted, failing to fully realize its potential in functional food development and cardiovascular protection. Furthermore, the lack of a comprehensive understanding of polysaccharides from different parts makes it difficult to determine whether the aerial parts can serve as a substitute or supplement to the roots, limiting the sustainable utilization of the entire chicory plant and hindering the wider and more efficient application of chicory polysaccharides in the food and nutritional supplement industry. Summary of the Invention

[0005] The purpose of this application is to overcome the above-mentioned technical problems and provide a method for extracting polysaccharides from the aerial parts of chicory, as well as the polysaccharides and related applications and evaluation methods.

[0006] In a first aspect, this application provides a method for extracting polysaccharides from the aerial parts of chicory, comprising the following steps: (1) Take the powdered aerial parts of chicory, add distilled water for reflux extraction, the extraction temperature is 80-100℃, the extraction time is 1-3 hours each time, and the extraction is repeated 1-3 times in total; (2) Combine the extracts and concentrate them under reduced pressure; (3) Add 60%-80% ethanol by volume to the concentrate and let it stand overnight at 0-10℃ to precipitate polysaccharides; (4) Centrifuge at 3000-5000 rpm for 5-15 minutes to collect the precipitate, and wash with anhydrous ethanol 2-6 times to remove impurities; (5) The washed precipitate was freeze-dried at -70℃ to -90℃ to obtain chicory aerial polysaccharide (CP-A).

[0007] By employing the above-mentioned technical solutions, polysaccharide components in chicory aerial part powder samples can be extracted using distilled water reflux under specific temperature, time, and number of cycles, effectively improving the extraction efficiency of polysaccharides. Concentrating the extract under reduced pressure reduces the solution volume, resulting in a more concentrated polysaccharide composition. Adding a specific volume fraction of ethanol and allowing it to stand overnight at low temperature selectively precipitates the polysaccharides, achieving good separation. Centrifugation at appropriate speeds and repeated washing with anhydrous ethanol effectively removes impurities and improves the purity of the polysaccharides. Freeze-drying at low temperature maximizes the preservation of the polysaccharide's biological activity, ultimately yielding high-purity, high-activity chicory aerial part polysaccharide (CP-A), providing a high-quality raw material basis for subsequent research and application.

[0008] Preferably, in step (1), the aboveground part of chicory is dried stem and leaf tissue, the mass of the powder sample is 20-40g, the volume of distilled water is 150-250mL, and the material-to-liquid ratio is 1g:(5-8)mL.

[0009] Preferably, the yield of the obtained chicory aerial polysaccharide (CP-A) is 12%-18%.

[0010] Preferably, the method further includes analyzing the molecular weight distribution of the obtained chicory aerial polysaccharide (CP-A) using a gel permeation chromatography (GPC) system equipped with a differential refractive index detector, a PLAquagel-OHMIXED-M (7.5×300mm) column or an equivalent column, a mobile phase of distilled water, a flow rate of 0.8-1.2 mL / min, a column temperature of 35-45℃, and using linear polyethylene glycol (PEG) as a standard to determine the molecular weight distribution range of 10kDa-1200kDa.

[0011] Preferably, the method further includes analyzing the monosaccharide composition of the obtained chicory aerial polysaccharide (CP-A) using a low-temperature hydrolysis method (50-70℃, 1-3M trifluoroacetic acid, 0.5-2 hours) and / or a conventional acid hydrolysis method (110-130℃, 1-3M trifluoroacetic acid, 1-3 hours), and using an ion chromatography system for separation and quantification. The monosaccharide composition includes arabinose, galactose, galacturonic acid, glucose, mannose, fructose, and rhamnose, and the fructose content accounts for more than 40% of the total sugar molar percentage.

[0012] Secondly, this application provides a chicory aerial polysaccharide (CP-A) extracted by the above-described method, which is extracted by any one of claims 1-5. Its weight-average molecular weight ranges from 50kDa to 800kDa, and its monosaccharide composition includes fructose, glucose, galactose, arabinose, galacturonic acid, rhamnose, and mannose, wherein the glucose content accounts for 20%-40% of the total monosaccharides, the fructose content accounts for 40%-60% of the total monosaccharides, and the galactose content accounts for 5%-25% of the total monosaccharides.

[0013] By adopting the above-mentioned technical solution and using a specific extraction method, chicory aerial polysaccharide was obtained. This polysaccharide has a specific weight-average molecular weight range and monosaccharide composition, which provides a basis for further research on its biological activity and application. Moreover, this polysaccharide has potential value in the preparation of products that protect cardiomyocytes, reduce myocardial oxidative damage, and inhibit myocardial inflammation.

[0014] Preferably, the polysaccharide has antioxidant activity, can scavenge free radicals, regulate intracellular redox homeostasis, and exert anti-inflammatory effects by inhibiting the MAPK / NF-κB signaling pathway, specifically by reducing the ratio of phosphorylated p38MAPK (Thr180 / Tyr182) to p38MAPK, reducing the ratio of phosphorylated NF-κBp65 (Ser536) to NF-κBp65, and downregulating the expression of MyD88 and CEBPB proteins.

[0015] Thirdly, this application provides the use of the above-mentioned chicory aerial polysaccharide (CP-A) in the preparation of products for protecting cardiomyocytes, reducing myocardial oxidative damage and / or inhibiting myocardial inflammation.

[0016] Preferably, the protection of cardiomyocytes or reduction of myocardial oxidative damage includes: treating H9c2 cells at a concentration of 10-200 μg / mL in an oxygen-glucose deprivation / reperfusion (OGD / R) model to increase cell viability by 30%-50% and reduce the levels of creatine kinase isoenzyme (CK-MB), cardiac troponin T (cTnT), and 8-hydroxy-2'-deoxyguanosine (8-OHdG).

[0017] Preferably, the inhibition of myocardial inflammation includes: downregulating the protein expression levels of myeloid differentiation factor 88 (MyD88) and CCAAT / enhancer-binding protein β (CEBPB) in cardiomyocytes, and / or inhibiting the phosphorylation of nuclear factor κBp65 subunit (NF-κBp65) and p38 mitogen-activated protein kinase (p38MAPK).

[0018] Preferably, the product is a drug, functional food, nutritional supplement or food additive, used to prevent or alleviate myocardial oxidative damage related to cardiovascular disease, wherein the amount of chicory aerial polysaccharide (CP-A) added is 0.1%-10.0% (w / v).

[0019] Fourthly, this application provides a myocardial cell protection composition comprising an effective amount of chicory aerial polysaccharide (CP-A) as described in claim 6 or 7 as an active ingredient, and a pharmaceutically or food-grade acceptable carrier or excipient.

[0020] By employing the above-mentioned technical solution, chicory aerial polysaccharide (CP-A) as an active ingredient can exert antioxidant activity, scavenge free radicals, regulate intracellular redox homeostasis, and exert anti-inflammatory effects by inhibiting the MAPK / NF-κB signaling pathway, reducing the ratio of phosphorylated p38MAPK to p38MAPK, reducing the ratio of phosphorylated NF-κBp65 to NF-κBp65, and downregulating the expression of MyD88 and CEBPB proteins. In the oxygen-glucose deprivation / reperfusion (OGD / R) model, treatment of H9c2 cells at a certain concentration can improve cell viability and reduce the levels of creatine kinase isoenzymes, cardiac troponin T, and 8-hydroxy-2'-deoxyguanosine. When combined with pharmaceutically or food-grade carriers or excipients, the composition can be used to protect cardiomyocytes, reduce myocardial oxidative damage, and inhibit myocardial inflammation.

[0021] Preferably, it further comprises chicory root polysaccharide (CP-R), wherein the mass ratio of CP-A to CP-R is 1:20-20:1, wherein the CP-R is prepared from chicory roots by the same extraction method as CP-A, and CP-A and CP-R are equivalent in cardiomyocyte protective activity, showing no significant difference in protective effect against OGD / R-induced H9c2 cardiomyocyte damage at the same concentration (p>0.05).

[0022] Preferably, the composition is a pharmaceutical composition, a health food, or a functional food additive, used to synergistically exert antioxidant and anti-inflammatory effects.

[0023] Fifthly, this application provides an evaluation method for comparing the cardioprotective activity of polysaccharides from different parts of chicory, comprising the following steps: (i) Polysaccharides are extracted from the roots and aerial parts of chicory using the method described in any one of claims 1-5 to obtain root polysaccharide (CP-R) and aerial polysaccharide (CP-A). (ii) Establish an oxygen-glucose deprivation / reperfusion (OGD / R) injury model of H9c2 cardiomyocytes; (iii) Treat the cells in the injury model with CP-R and CP-A at concentrations of 10-200 μg / mL, respectively; (iv) The cardioprotective activity of CP-R and CP-A was evaluated and compared by detecting cell viability, myocardial injury markers (including CK-MB and cTnT) and oxidative stress markers (including 8-OHdG). (v) The protein expression levels of MyD88, CEBPB, phosphorylated NF-κBp65 and phosphorylated p38MAPK in cardiomyocytes were detected by Western blotting to elucidate the molecular mechanisms of CP-R and CP-A. If CP-A exhibits protective activity comparable to CP-R, then the aerial parts of chicory can be considered as an alternative source of polysaccharides with cardioprotective activity.

[0024] By adopting the above-mentioned technical solutions, the cardioprotective activities of chicory root polysaccharides and aerial polysaccharides can be systematically and comprehensively compared, clarifying the differences in their effects on protecting cardiomyocytes, reducing myocardial oxidative damage, and inhibiting myocardial inflammation. The molecular mechanisms by which the two polysaccharides exert their cardioprotective effects can be explored in depth. If the aerial polysaccharides of chicory exhibit protective activity comparable to that of the root polysaccharides, it can be determined that the aerial parts of chicory can serve as an alternative source of polysaccharides with cardioprotective activity, providing a basis for the comprehensive utilization and development of chicory resources.

[0025] In summary, this application includes at least one of the following beneficial technical effects: 1. Polysaccharides can be extracted from the aerial parts of chicory using a specific extraction method, with a yield of 12%-18%. This effectively utilizes the previously discarded aerial parts of chicory, turning waste into treasure and achieving high resource efficiency. For the first time, the traditionally discarded aerial parts of chicory (stems and leaves) have been developed into raw materials for preparing polysaccharides with clear cardioprotective activity. This realizes the sustainable and high-value utilization of the entire chicory plant resource, reduces raw material costs, and has significant economic and environmental implications.

[0026] 2. Through rigorous cell modeling (OGD / R-induced H9c2 cell damage), the chicory aerial polysaccharide (CP-A) provided in this application has been demonstrated to possess significant antioxidant, anti-inflammatory, and cardioprotective effects, comparable to the activity of the recognized root polysaccharide (CP-R). Furthermore, multi-omics techniques have elucidated that its protective effect is achieved through the inhibition of the MAPK / NF-κB signaling pathway, providing a solid scientific basis for product development.

[0027] Chicory aerial polysaccharides can downregulate the protein expression levels of myeloid differentiation factor 88 and CCAAT / enhancer-binding protein β in cardiomyocytes, inhibit phosphorylation of nuclear factor κB p65 subunit and p38 mitogen-activated protein kinase, and suppress myocardial inflammation. Chicory aerial polysaccharides can be used to prepare pharmaceuticals, functional foods, and other products to prevent or alleviate cardiovascular disease-related myocardial oxidative damage. They can also synergistically exert antioxidant and anti-inflammatory effects when combined with chicory root polysaccharides. Furthermore, evaluation methods can be used to determine whether chicory aerial parts can serve as an alternative source of polysaccharides with cardioprotective activity, promoting the sustainable utilization of the entire chicory plant. Based on the well-defined bioactivity of CP-A, it can be developed into drugs, health foods, functional foods, or food additives for protecting cardiomyocytes and preventing or adjuvantly treating cardiovascular diseases, with significant market potential.

[0028] The activity evaluation method established in this application provides an objective and quantifiable technical standard for determining whether the aboveground parts of chicory can be used as a substitute for the root (e.g., no statistically significant difference in protective effect), which has guiding significance for the selection of raw materials and quality control of the entire industry. Attached Figure Description

[0029] Figure 1 Morphological characteristics and raw materials of chicory. Among them: (A) Fresh whole chicory plant harvested from the cultivation field, showing the above-ground parts and roots; (B) Dried aerial parts of chicory used to prepare CP-A; (C) Dried chicory roots used to prepare CP-R.

[0030] Figure 2 Effects of chicory aerial parts (CP-A) and root polysaccharides (CP-R) on H9c2 cell viability and damage biomarkers under OGD / R conditions. (The figure shows the effects of these polysaccharides on H9c2 cell viability and damage biomarkers under OGD / R conditions.) (A,B) Effects of different concentrations (0-1000 μg / mL) of CP-A and CP-R on H9c2 cell viability after 24 hours of treatment; (C) Effects of OGD / R intervention and different concentrations of CP-A (CP-AL, 25 μg / mL; CP-AM, 50 μg / mL; CP-AH, 100 μg / mL) and CP-R (CP-RL, 25 μg / mL; CP-RM, 50 μg / mL; CP-RH, 100 μg / mL) on H9c2 cell viability. Cell viability was determined using the CCK-8 assay. (DF) Effects of OGD / R intervention and high-dose CP-A (CP-AH, 100 μg / mL) or CP-R (CP-RH, 100 μg / mL) on the levels of (D) creatine kinase-MB (CK-MB), (E) cardiac troponin T (cTnT) and (F) 8-hydroxydeoxyguanosine (8-OHdG) in H9c2 cells.

[0031] Figure 3 Proteomic analysis of differentially expressed proteins and enriched pathways in the control group, model group, and CP-treated group. Among them: (A) Volcano plot of differentially expressed proteins between the model group and the control group; (B) KEGG pathway enrichment analysis of differentially expressed proteins between the model group and the control group; (C) Volcano plot of differentially expressed proteins between the CP-A group and the model group; (D) KEGG pathway enrichment analysis of differentially expressed proteins between the CP-A group and the model group; (E) Volcano plot of differentially expressed proteins between the CP-R group and the model group; (F) KEGG pathway enrichment analysis of differentially expressed proteins between the CP-R group and the model group.

[0032] Figure 4 Overlap analysis and Gene Ontology (GO) enrichment analysis of differentially expressed proteins. Among them: (A) Venn diagrams show the intersection of differentially expressed proteins in the comparisons between the model group vs. control group, the CP-A group vs. the model group, and the CP-R group vs. the model group; (B) GO enrichment analysis of 398 overlapping proteins, divided into biological processes (BP), cellular components (CC), and molecular functions (MF).

[0033] Figure 5 Transcriptomic analysis and pathway enrichment analysis of CP-A and CP-R treatment in H9c2 cells. Among them: (A) Volcano plot of differentially expressed genes (DEGs) between the CP-A group and the model group; (B) KEGG pathway enrichment analysis of differentially expressed genes between the CP-A group and the model group; (C) Gene set enrichment analysis (GSEA) showed that MAPK and Toll-like receptor signaling pathways were the most significantly enriched biological processes in the CP-A group; (D) Volcano diagram of differentially expressed genes between the CP-R group and the model group;

[0034] (E) KEGG pathway enrichment analysis of differentially expressed genes between the CP-R group and the model group; (F) Gene set enrichment analysis (GSEA) showed that the MAPK and NF-κB signaling pathways were the most significantly enriched pathways in the CP-R group.

[0035] Figure 6 .Analysis of downregulated genes and GO functional enrichment common to CP-A and CP-R treatments. Among them: (A) Venn diagrams show overlapping downregulated differentially expressed genes (DEGs) in the comparison of CP-A group vs. model group and CP-R group vs. model group; (B) GO enrichment analysis of 290 intersecting genes, classified by biological process (BP), cellular component (CC) and molecular function (MF); (C) Cytoscape visualization of GO functional networks based on biological process enrichment, highlighting signaling connections centered on MAPK.

[0036] Figure 7 Validation of the regulation of the MAPK / NF-κB pathway by CP-A and CP-R in H9c2 cells. Among them: (A) Expression of p-P65NF-κB, P65NF-κB, MyD88, p-P38MAPK, P38MAPK and CEBPB proteins as detected by Western blot; Quantitative analysis of the expression levels of (BE) p-P65 / P65, MyD88 / β-actin, p-P38 / P38 and CEBPB / β-actin proteins. Detailed Implementation

[0037] The following embodiments are provided to further illustrate this application, but do not constitute any limitation on the scope of protection of this application. Those skilled in the art can make various modifications or substitutions without departing from the spirit and substance of this application, and these modifications or substitutions should all be covered within the scope of protection of this application.

[0038] This application aims to address the technical problem of resource waste in the aboveground parts of chicory and provide a novel and substantially progressive utilization approach. Through systematic research, this application reveals and confirms for the first time that although the polysaccharide extracted from the aboveground parts of chicory (CP-A) differs measurably from the traditional root polysaccharide (CP-R) in chemical characteristics such as molecular weight and monosaccharide ratio, both exhibit statistically indistinguishable protective effects in an oxygen-glucose deprivation / reperfusion (OGD / R)-induced cardiomyocyte injury model. Further in-depth multi-omics mechanism studies show that this functional equivalence stems from their combined action on the core stress-inflammatory signaling pathway MAPK / NF-κB in cardiomyocytes, synergistically inhibiting it. This discovery breaks with the traditional understanding that only roots are a source of active polysaccharides, laying a solid scientific foundation for the high-value resource utilization of the aboveground parts of chicory, and constituting the core inventiveness of this application. Example 1

[0039] Standardized preparation, yield optimization and basic chemical characterization of polysaccharides from chicory aerial parts and roots To ensure the accuracy and fairness of subsequent bioactivity comparisons and to optimize extraction efficiency, different extraction parameters were first screened, and a final standardized extraction procedure was established for the parallel preparation of CP-A and CP-R. This was combined with the attached... Figure 1 illustrate.

[0040] The chicory (Cichorium intybus L.) used in the experiment was purchased from Hebei Fengning Ping An High-Tech Industry Co., Ltd. The above-ground parts (stems and leaves) and roots of the fresh plants were separated, washed, and dried separately at 50℃ to constant weight. The dried powder was then pulverized and passed through a 60-mesh sieve for later use.

[0041] In optimizing the extraction process, we systematically investigated the effects of the material-to-liquid ratio, the number of extractions, and the ethanol concentration on the polysaccharide yield. For the aerial parts, when the extraction temperature was fixed at 95-100℃ and each extraction lasted 2 hours, increasing the material-to-liquid ratio from 1:5 to 1:8 increased the yield from 13.2% to 15.1%. However, further increasing the water ratio to 1:10 did not significantly improve the yield (15.3%). Therefore, a ratio of 1:6.67 (i.e., 30g raw material / 200mL water) was selected as the optimal condition for balancing efficiency and cost. The extraction number experiment showed that the yield was 10.5% with one extraction, 14.8% with two extractions, and only slightly increased to 15.0% with three extractions. Therefore, two extractions were determined to be the optimal number. The ethanol precipitation concentration experiment showed that the yield was low and the precipitate was loose when 60% ethanol was used for precipitation, while 70% ethanol resulted in complete precipitation and the highest yield. The yield decreased slightly and the co-precipitation of impurities increased when 80% ethanol was used. Therefore, 70% ethanol was selected as the optimal precipitation concentration.

[0042] Based on the above optimization, 30.0 g each of the above-mentioned aerial part powder and root powder were accurately weighed and placed in separate 1000 mL round-bottom flasks. 200 mL of distilled water was added to each flask, and a reflux condenser was connected. The mixture was heated to a gentle boil (temperature maintained at 95-100℃) and timing was started, followed by reflux extraction for 2 hours. After the first extraction, the mixture was filtered while hot, and the filtrate was collected. The residue was returned to the flask, and another 200 mL of distilled water was added. The reflux extraction process was repeated for 2 hours. The filtrates from the two extractions were combined.

[0043] The combined extracts were concentrated to approximately 50 mL (about 12.5% ​​of the original volume) by rotary evaporation under reduced pressure at -0.085 MPa in a 55°C water bath. The concentrate was then placed in an ice-water bath at 4°C, and pre-cooled anhydrous ethanol was added dropwise at approximately 2 mL / min with slow stirring until the final ethanol volume fraction reached 70%. After the addition was complete, the mixture was allowed to stand at 4°C overnight (12-16 hours) to precipitate. The next day, the precipitate mixture was centrifuged at 4000 × g for 15 minutes, and the precipitate was collected. The precipitate was washed three times (20 mL each time) with pre-cooled anhydrous ethanol to remove residual pigments and small molecule impurities. The washed precipitate was then frozen at -80°C for 4 hours, transferred to a freeze dryer, and freeze-dried for 48 hours at a condenser temperature < -50°C and a vacuum < 10 Pa to obtain white chicory aerial polysaccharide (CP-A) and root polysaccharide (CP-R) freeze-dried powders.

[0044] After five independent repeated extraction experiments, the average yield of CP-A was 14.8% ± 0.5% (based on the mass of dried raw material), and the average yield of CP-R was 15.3% ± 0.4%. Routine quality analysis of the freeze-dried products showed that the protein residue (determined by the BCA method) of CP-A was less than 0.5%, and the ash content (550℃ ignition method) was less than 2.0%; the corresponding indicators for CP-R were less than 0.7% and less than 2.5%, respectively. These data indicate that the polysaccharide yield from the aerial parts of chicory is comparable to that from the roots, and the purity is good, providing a basis for its use as a substitute raw material in terms of both yield and quality.

[0045] To objectively characterize the similarities and differences in physicochemical properties between the two, we conducted a systematic structural analysis. Molecular weight distribution was determined using gel permeation chromatography. The sample was prepared as a 5 mg / mL aqueous solution, filtered through a 0.22 μm filter membrane, and then injected. The system was a Waters Alliance e2695, the column was a Plaquagel-OHMIXED-M (7.5 × 300 mm, 8 μm), the mobile phase was ultrapure water, the flow rate was 1.0 mL / min, the column temperature was 40 °C, and a differential refractive index detector was used for detection. A standard curve was plotted using polyethylene glycol (PEG) standards (1, 5, 12, 50, 150, 400, 800 kDa) (lgMw = -0.321t + 11.62, R² = 0.998). The results showed that the chromatogram of CP-A showed a main peak retention time of 14.2 minutes, with a calculated weight-average molecular weight (Mw) of 352±15 kDa, a number-average molecular weight (Mn) of 210±10 kDa, and a dispersion index (Đ, Mw / Mn) of 1.68±0.05. In contrast, the main peak retention time of CP-R was 13.5 minutes, with a Mw of 680±25 kDa, a Mn of 310±12 kDa, and an Đ of 2.19±0.08. This indicates that the molecular weight distribution of CP-A is more concentrated than that of CP-R, and the average molecular weight is lower, suggesting an inherent difference in molecular size between the two.

[0046] The monosaccharide composition was further analyzed by ion chromatography. 3.0 mg of sample was accurately weighed, and 1 mL of 2M trifluoroacetic acid (TFA) was added. After nitrogen purging and sealing, the mixture was hydrolyzed in an oil bath at 121 °C for 2 hours. The hydrolysate was dried under nitrogen, washed twice with methanol to remove acid, and then reconstituted with 1 mL of ultrapure water and filtered through a 0.22 μm filter membrane. A ThermoICS-5000+ system was used, with a Dionex CarboPac™ PA20 analytical column (3 × 150 mm) and an electrochemical detector (pulse amperometry, gold electrode). The mobile phase consisted of ultrapure water (A), 100 mM NaOH (B), and 1 M NaOAc (C), using gradient elution: 0–20 min, 2% B isocratic; 20–25 min, B increased from 2% to 100%; 25–30 min, 100% B + 5% C; 30–35 min, 100% B + 20% C. The flow rate was 0.5 mL / min, and the column temperature was 30 °C. Calibration curves were plotted using seven monosaccharide standards (concentration gradient: 0.5, 1, 2, 5, 10 μg / mL). Each standard showed good linearity within the selected concentration range (R² > 0.995). The results indicated that both CP-A and CP-R are composed of seven monosaccharides: fructose (Fru), glucose (Glc), galactose (Gal), arabinose (Ara), galacturonic acid (GalA), rhamnose (Rha), and mannose (Man), but their molar proportions differed significantly. The average results of three parallel determinations showed that the molar percentages of fructose, glucose, and galactose in CP-A were 52.3 ± 1.5%, 28.5 ± 0.9%, and 12.4 ± 0.6%, respectively; while in CP-R, these proportions were 58.7 ± 1.8%, 22.1 ± 0.7%, and 10.5 ± 0.5%, respectively. The contents of the remaining monosaccharides are shown in Table 1. This confirms at the chemical composition level that CP-A is a chicory polysaccharide that is structurally similar to but distinct from CP-R.

[0047] Table 1: Monosaccharide molar composition of CP-A and CP-R (%, mean ± standard deviation, n=3) Example 2

[0048] Comparative evaluation of the protective activities of CP-A and CP-R in a cardiomyocyte injury model and their dose-response relationship Having clarified the structural differences between the two, the core of this embodiment lies in systematically evaluating the similarities and differences in their biological functions and dose-response relationship. We used the H9c2 rat cardiomyocyte line to establish an oxygen-glucose deprivation / reperfusion (OGD / R) model to simulate myocardial ischemia-reperfusion injury. And combined with the attached... Figure 2 illustrate.

[0049] Cells were cultured in DMEM high-glucose medium (HyClone) containing 10% fetal bovine serum (FBS, Gibco) and placed in a saturated humidity incubator at 37°C and 5% CO2. When cells reached 70-80% confluence, OGD / R modeling was initiated. First, cells were washed twice with preheated glucose-free DMEM (purged with a 95% N2 / 5% CO2 mixture for 30 minutes), and then glucose-free DMEM was added. The cell culture plates were placed in a sealed anaerobic incubator (Mitsubishi Gas Chemical) filled with a 95% N2 and 5% CO2 mixture and incubated at 37°C for 2 hours to induce oxygen-glucose deprivation (OGD). Subsequently, the medium was rapidly replaced with normal complete medium (containing glucose and 10% FBS), and the cells were returned to a normoxic (95% air, 5% CO2) incubator for another 24 hours to simulate the reperfusion process. At the start of reperfusion, different concentrations of CP-A or CP-R were added to the culture medium for treatment. The following experimental groups were established: ① Normal control group (Control, normoxic culture); ② OGD / R model group (Model, modeling, no polysaccharide added); ③ CP-A treatment group (25, 50, 100 μg / mL); ④ CP-R treatment group (25, 50, 100 μg / mL). All treatment groups had six replicates after modeling.

[0050] Cell viability was first assessed using the CCK-8 assay. Twenty-four hours after reperfusion, 10 μL of CCK-8 reagent (Dojindo) was added to each well, and the cells were incubated at 37°C in the dark for 2 hours. The absorbance (OD value) at 450 nm was measured using a BioTek microplate reader. Cell viability (%) = [(OD treatment group - OD blank) / (OD normal control group - OD blank)] × 100%. Preliminary cytotoxicity tests showed that CP-A and CP-R had no significant effect on the viability of H9c2 cells (viability > 95%) after incubation for 24 hours within a concentration range of 0-1000 μg / mL, indicating their safety at the experimental concentrations.

[0051] The results of the cell viability assay are shown in the figure. Compared with the normal control group (viability set at 100%), the OGD / R model resulted in a significant decrease in cell viability to 55.2% ± 3.1%. Both CP-A and CP-R treatments reversed this damage in a concentration-dependent manner. At concentrations of 25, 50, and 100 μg / mL, the cell viability in the CP-A group was 68.5% ± 4.2%, 82.1% ± 3.8%, and 94.7% ± 2.5%, respectively; while in the CP-R group it was 66.8% ± 3.9%, 80.5% ± 4.1%, and 93.1% ± 3.0%, respectively. One-way ANOVA and Tukey's multiple comparison test were performed using GraphPadPrism 9.0 software. Statistical analysis showed that there was no significant difference in cell viability between the CP-A treatment group and the CP-R treatment group at each corresponding concentration point (25, 50, 100 μg / mL) (p>0.05); however, there were highly significant differences between each dose group of the two polysaccharides and the OGD / R model group (p<0.001). The half-maximal effect concentrations (COPs) of CP-A and CP-R were calculated by fitting the dose-response curves using nonlinear regression. 50 The concentrations were 38.2 ± 2.1 μg / mL and 40.1 ± 2.5 μg / mL, respectively, with no statistically significant difference between the two. This result provides the most direct and quantitative evidence for the "activity equivalence" between the two.

[0052] To further quantify the degree of myocardial injury and oxidative stress, we detected myocardial injury markers creatine kinase isoenzyme MB (CK-MB) and cardiac troponin T (cTnT) in cell culture supernatant, and oxidative DNA damage marker 8-hydroxy-2'-deoxyguanosine (8-OHdG) in cell lysate. Commercial kits from Nanjing Jiancheng Biotechnology Institute were used, strictly following the instructions. CK-MB and cTnT activities / contents were determined using a microplate reader, and 8-OHdG content was determined using a competitive ELISA method.

[0053] The results are as follows Figure 2As shown in the figure, after OGD / R injury, compared with the normal control group, the model group showed an approximately 3.8-fold increase in CK-MB activity, an approximately 4.2-fold increase in cTnT content, and an approximately 5.1-fold increase in intracellular 8-OHdG level. Treatment with 100 μg / mL CP-A or CP-R significantly inhibited the levels of these three biomarkers. Specifically, for 8-OHdG, with the model group level set at 100%, the CP-AH group decreased to 42.3% ± 5.1%, and the CP-RH group decreased to 45.1% ± 4.8%. For CK-MB, the CP-AH and CP-RH groups decreased to 48.5% ± 6.2% and 50.1% ± 5.7% of the model group, respectively. For cTnT, the levels decreased to 51.8% ± 4.9% and 53.2% ± 5.4%, respectively. Statistical analysis showed no significant difference in inhibitory effects between the CP-AH and CP-RH groups for any of the biomarkers (p>0.05). These data consistently confirm, across multiple independent biochemical indicators, that CP-A has the same efficacy as CP-R in reducing damage to myocardial cell membrane integrity, myofibril damage, and DNA oxidative damage. Example 3

[0054] Molecular mechanism exploration and common target analysis based on proteomics and transcriptomics To elucidate the common molecular basis for the equivalent protective effects of CP-A and CP-R, we went beyond phenotypic observation and integrated proteomics and transcriptomics technologies to perform a global, non-targeted analysis of H9c2 cells after OGD / R damage and polysaccharide treatment. This was combined with... Figure 3-7 illustrate.

[0055] Quantitative proteomics analysis (LC-MS / MS) using TMT labeling was performed on cell samples from the normal control group, OGD / R model group, CP-AH group (100 μg / mL), and CP-RH group (100 μg / mL), with three biological replicates for each group. Mass spectrometry data were processed using ProteomeDiscoverer 2.4 software, and the Uniprot rat database was searched. The screening criteria for differentially expressed proteins were: fold change >1.5 or <0.67, and p-value <0.05 (Student's t-test). As shown in the figure, compared with the control group, a total of 1524 differentially expressed proteins were identified (706 downregulated and 818 upregulated). KEGG pathway enrichment analysis showed that these proteins were significantly enriched in inflammation and stress-related pathways such as the p38MAPK signaling pathway (p=3.2E-7), the TNF signaling pathway (p=1.1E-6), and the NF-κB signaling pathway (p=4.5E-6), as well as pathways such as oxidative phosphorylation (p=2.8E-8) and apoptosis (p=7.3E-5), comprehensively reflecting the complex cellular stress response induced by OGD / R.

[0056] Crucially, CP-A treatment (vs. model group) reversed the expression of 1210 proteins (784 downregulated, 426 upregulated), with enriched pathways including oxidative stress response (p=2.1E-5) and IL-2 signaling (p=1.8E-4). CP-R treatment (vs. model group) reversed the expression of 1058 proteins (632 downregulated, 426 upregulated), also significantly enriched in the p38MAPK signaling pathway (p=5.6E-6) and NF-κB signaling pathway (p=9.3E-5). Venn diagram analysis revealed that CP-A and CP-R treatments jointly regulated 398 overlapping differentially expressed proteins. Gene Ontology (GO) enrichment analysis of these 398 core intersection proteins revealed that they are primarily involved in biological processes such as aerobic respiration (p=4.3E-9), oxidative stress response (p=6.7E-8), and angiogenesis (p=2.1E-5), and are enriched in molecular functions such as antioxidant activity (p=1.5E-6) and oxidoreductase activity (p=3.8E-5). This indicates that, despite their structural differences, CP-A and CP-R intervene in a highly overlapping protein network related to cellular redox homeostasis and energy metabolism.

[0057] At the transcriptional level, RNA-Seq analysis (Illumina NovaSeq platform, ≥40 Mclean reads per sample) was performed on the four cell groups. Differentially expressed genes (DEGs) were selected based on |log2FC|>1 and FDR<0.05. Both CP-A and CP-R treatments significantly regulated the expression of hundreds of genes. KEGG enrichment analysis and gene set enrichment analysis (GSEA) consistently showed that both significantly affected the MAPK signaling pathway, the Toll-like receptor signaling pathway, and the NF-κB signaling pathway. Importantly, the analysis revealed that CP-A and CP-R jointly downregulated 290 genes. GO analysis of these 290 intersecting genes showed significant enrichment in categories such as protein phosphorylation (p=8.9E-12), negative regulation of the MAPK cascade (p=3.4E-9), and cellular response to oxidative stress (p=1.2E-7). Visualizing biological process networks using Cytoscape software clearly reveals that the MAPK signaling pathway is located at the center of this regulatory network and is closely connected with the Ras signaling pathway, calcium signaling pathway, and others. This strongly demonstrates at the gene transcription level that the protective effects of CP-A and CP-R converge on the inhibition of a key stress signaling network centered on MAPK.

[0058] To validate the predictions of the above omics analysis, we used Western blotting to detect the expression and activation status of key proteins in the MAPK / NF-κB pathway at the protein function level. Total cellular protein was collected, and its concentration was determined using the BCA method. An equal amount of protein was subjected to SDS-PAGE electrophoresis and transferred to a PVDF membrane. After blocking, the gels were incubated overnight at 4°C with the following primary antibodies: phosphorylated p38MAPK (Thr180 / Tyr182) (1:1000, ZenBioScience), p38MAPK (1:1000, ZenBioScience), phosphorylated NF-κBp65 (Ser536) (1:1000, ZenBioScience), NF-κBp65 (1:1000, ZenBioScience), MyD88 (1:1000, Proteintech), CEBPB (1:1000, ZenBioScience), and β-actin (1:5000, Proteintech). Subsequently, the gels were incubated with HRP-labeled secondary antibodies, ECL was developed, images were acquired using the Syngene gel imaging system, and grayscale values ​​were quantified using ImageJ software.

[0059] The results are shown in the figure. OGD / R damage significantly increased the levels of phosphorylated p38MAPK (p-p38) and phosphorylated NF-κBp65 (p-p65) (their ratios to total protein increased by approximately 3.5-fold and 4.2-fold, respectively, compared to the control group), while upregulating the protein expression of MyD88 and CEBPB (increasing by approximately 2.8-fold and 3.1-fold, respectively). After treatment with 100 μg / mL CP-A or CP-R, the p-p38 / p38 ratio decreased to 0.39±0.04 and 0.36±0.05 in the model group, respectively; the p-p65 / p65 ratio decreased to 0.41±0.05 and 0.38±0.06, respectively; the MyD88 / β-actin ratio decreased to 0.52±0.07 and 0.48±0.06, respectively; and the CEBPB / β-actin ratio decreased to 0.55±0.05 and 0.51±0.06, respectively. Statistical analysis (ANOVA, Tukey's test) showed no significant difference in the degree of inhibition between the CP-AH group and the CP-RH group for each detected protein index (p>0.05). This result was ultimately confirmed at the protein function level: CP-A and CP-R exert equivalent cardioprotective effects by synergistically inhibiting the core inflammatory-stress signaling axis MyD88 / MAPK / NF-κB / CEBPB. Example 4

[0060] Product application examples and stability tests based on the principle of activity equivalence. Based on the above scientific conclusion of "activity equivalence", chicory aerial polysaccharide (CP-A) can be used alone or in combination with root polysaccharide (CP-R) to develop products with cardioprotective effects, and its application forms are diverse.

[0061] Application Example 1: Preparation and Preliminary Stability of CP-A Health Capsules The lyophilized CP-A powder prepared in Example 1 was passed through an 80-mesh sieve and accurately weighed according to the prescription (each capsule contains 250 mg of CP-A, 145 mg of microcrystalline cellulose, and 5 mg of magnesium stearate). All components were mixed in a three-dimensional mixer at 30 rpm for 30 minutes. The mixture was then filled into size 0 hard capsules using a fully automated capsule filling machine (e.g., model ZJT-40). Preliminary accelerated stability tests were conducted on the filled capsules: they were placed in a constant temperature and humidity chamber at 40℃±2℃ and 75%±5% relative humidity, and samples were taken at the end of 0, 1, 2, and 3 months. The results showed that after 3 months, the capsule appearance remained unchanged, the CP-A content (determined by the phenol-sulfuric acid method) remained above 98.5% of the initial content, and the microbial limits met the requirements of the General Rules of the Chinese Pharmacopoeia, indicating that CP-A has good stability in solid dosage forms.

[0062] Application Example 2: Formulation and Sensory Evaluation of CP-A / CP-R Compound Functional Solid Beverages A compound chicory polysaccharide solid beverage was designed. The formula is as follows: compound chicory polysaccharide (CP-A:CP-R=1:1, mass ratio) 10.0%, resistant dextrin (dietary fiber) 30.0%, erythritol (sweetener) 56.5%, natural lemon powder (flavoring agent) 2.0%, citric acid (acidity regulator) 1.0%, and sucralose (high-intensity sweetener) 0.5%. All powdered ingredients were mixed in a double cone mixer for 45 minutes until homogeneous. A simple sensory evaluation was conducted: 10g of the mixed powder was dissolved in 200mL of 40℃ warm water, and the product was tasted by 10 trained evaluators. The results showed that the product dissolved rapidly, the solution was clear, the taste was pleasantly sweet and sour, with a pleasant lemon flavor and a slight chicory-specific aftertaste, and the overall acceptability was high. Example 5

[0063] Standardized evaluation method for raw material substitutability and its verification To ensure the industrial applicability of the technological achievements of this application and to establish reliable quality control standards, we have summarized a standardized, progressive evaluation process for scientifically determining whether the aerial parts of chicory from a specific source can replace the roots in terms of cardiomyocyte protective activity.

[0064] The method includes the following specific steps: Parallel preparation of test samples: Following the standard extraction method described in Example 1 (fixed material-to-liquid ratio 1:6.67, reflux at 95-100℃ for 2 hours twice, precipitation with 70% ethanol), root polysaccharides (CP-R, as an internal positive control) and aerial polysaccharides (CP-A, the sample to be evaluated) were simultaneously prepared from the same batch and origin of chicory raw materials. The extraction yields of both were recorded.

[0065] Parallel cell viability assay: The OGD / R model establishment method and cell culture conditions described in Example 2 were strictly followed. A test concentration of 100 μg / mL is recommended (this concentration showed near-maximum protective effect in Example 2). The experimental groups must include: ① Normal control group (Control); ② OGD / R model group (Model); ③ CP-R treatment group (100 μg / mL); ④ CP-A treatment group (100 μg / mL). Each group should have at least 6 biological replicate wells (n=6).

[0066] Core activity index detection and raw data recording: Measure and record the following data: Cell viability (CCK-8 assay, 450nm OD value) CK-MB activity in cell supernatant (U / L) cTnT content in cell supernatant (ng / mL) Intracellular 8-OHdG content (ng / mg protein) Data processing and statistical analysis: Use professional statistical software (such as SPSS or GraphPad Prism). First, perform one-way ANOVA on each group of data (usually conforming to a normal distribution). If the ANOVA shows a significant difference between groups (p<0.05), further perform multiple comparison tests, such as Tukey's HSD test or LSD test. Pay special attention to the comparison results between the CP-R group and the CP-A group on each indicator.

[0067] Equivalence criteria: If both of the following conditions are met simultaneously, the above-ground polysaccharide (CP-A) of this batch of chicory can be considered an equivalent substitute for its root polysaccharide (CP-R) in terms of cardioprotective activity: Condition A (Efficacy): The CP-A treatment group and the CP-R treatment group showed statistically significant differences from the OGD / R model group in the four core indicators of cell viability, CK-MB, cTnT, and 8-OHdG (p<0.05), and the numerical direction showed a protective effect (i.e., increased viability and decreased damage markers).

[0068] Condition B (equivalence): There were no statistically significant differences in the measurements of all four core indicators between the CP-A treatment group and the CP-R treatment group (p>0.05).

[0069] (Optional Enhanced Validation) Mechanism Consistency Check: To further increase the reliability of the conclusion, a portion of cell samples can be extracted, and the inhibitory effects of CP-A and CP-R treatments on p-p38MAPK phosphorylation levels can be compared using the Western Blot method described in Example 3. If the inhibitory trends of both treatments on key signaling molecules are consistent (i.e., both can significantly inhibit them, and there is no statistically significant difference in the degree of inhibition), then further evidence of "equivalence" is provided at the molecular mechanism level.

[0070] To verify the reliability of this evaluation method, chicory raw materials from three different origins were tested. Chicory from origins A and B showed CP-A and CP-R passing the aforementioned equivalence criteria for all four activity indicators (p>0.05). However, chicory from origin C showed no difference in cell viability between CP-A and CP-R, but its protective effect against 8-OHdG was significantly weaker than CP-R (p<0.05), and therefore it was deemed "not completely equivalent." This demonstrates that this evaluation method has the ability to distinguish between raw materials of different qualities.

[0071] Through the above specific, repeatable, and data-rich embodiments, this application fully demonstrates the entire process of developing high-value bioactive polysaccharides from waste aboveground parts, and uses multi-level and multi-dimensional experimental data to demonstrate its core finding—structurally different yet functionally equivalent, and this equivalence has a unified molecular mechanism basis. This provides an innovative and substantially technologically advanced technical solution and a clear and reliable implementation path for the sustainable utilization of chicory resources and the development of related functional products.

[0072] The above are all preferred embodiments of this application, and are not intended to limit the scope of protection of this application. Therefore, all equivalent changes made in accordance with the structure, shape and principle of this application should be covered within the scope of protection of this application.

Claims

1. A method for extracting polysaccharides from the aerial parts of chicory, characterized in that, Includes the following steps: (1) Take the powdered aerial parts of chicory, add distilled water for reflux extraction, the extraction temperature is 80-100℃, the extraction time is 1-3 hours each time, and the extraction is repeated 1-3 times; (2) Combine the extracts and concentrate them under reduced pressure; (3) Add 60%-80% ethanol to the concentrate and let it stand overnight at 0-10℃ to precipitate polysaccharides; (4) Centrifuge at 3000-5000 rpm for 5-15 minutes to collect the precipitate, and wash it with anhydrous ethanol 2-6 times to remove impurities; (5) Freeze-dry the washed precipitate at -70℃ to -90℃ to obtain chicory aerial part polysaccharide (CP-A).

2. The extraction method according to claim 1, characterized in that, In step (1), the aboveground parts of chicory are dried stem and leaf tissues, the mass of the powder sample is 20-40g, the volume of distilled water is 150-250mL, and the material-to-liquid ratio is 1g:(5-8)mL.

3. The extraction method according to claim 1 or 2, characterized in that, The yield of the obtained chicory aerial polysaccharide (CP-A) was 12%-18%.

4. The extraction method according to any one of claims 1-3, characterized in that, Further analysis included molecular weight distribution analysis of the obtained chicory aerial polysaccharide (CP-A) using a gel permeation chromatography (GPC) system equipped with a differential refractive index detector. The chromatographic column was a Plaquagel-OHMIXED-M (7.5×300mm) or equivalent column. The mobile phase was distilled water, the flow rate was 0.8-1.2 mL / min, and the column temperature was 35-45℃. Linear polyethylene glycol (PEG) was used as a standard, and the molecular weight distribution range was 10kDa-1200kDa.

5. The extraction method according to any one of claims 1-3, characterized in that, Further analysis included monosaccharide composition analysis of the obtained chicory aerial polysaccharide (CP-A) using a low-temperature hydrolysis method (50-70℃, 1-3M trifluoroacetic acid, 0.5-2 hours) and / or a conventional acid hydrolysis method (110-130℃, 1-3M trifluoroacetic acid, 1-3 hours), and separation and quantification using an ion chromatography system. The monosaccharide composition included arabinose, galactose, galacturonic acid, glucose, mannose, fructose, and rhamnose, with fructose content accounting for more than 40% of the total sugar molar percentage.

6. A chicory aerial part polysaccharide (CP-A), characterized in that, The product is extracted by the method of any one of claims 1-5, and its weight-average molecular weight ranges from 50kDa to 800kDa. The monosaccharide composition includes fructose, glucose, galactose, arabinose, galacturonic acid, rhamnose, and mannose, wherein the glucose content accounts for 20%-40% of the total monosaccharides, the fructose content accounts for 40%-60% of the total monosaccharides, and the galactose content accounts for 5%-25% of the total monosaccharides.

7. The chicory aerial part polysaccharide (CP-A) according to claim 6, characterized in that, The polysaccharide has antioxidant activity, which can scavenge free radicals, regulate intracellular redox homeostasis, and exert anti-inflammatory effects by inhibiting the MAPK / NF-κB signaling pathway. Specifically, it reduces the ratio of phosphorylated p38MAPK (Thr180 / Tyr182) to p38MAPK, reduces the ratio of phosphorylated NF-κBp65 (Ser536) to NF-κBp65, and downregulates the expression of MyD88 and CEBPB proteins.

8. The use of the chicory aerial polysaccharide (CP-A) according to claim 6 or 7 in the preparation of products for protecting cardiomyocytes, reducing myocardial oxidative damage and / or inhibiting myocardial inflammation.

9. The application according to claim 8, characterized in that, The protection of cardiomyocytes or reduction of myocardial oxidative damage includes: treating H9c2 cells at a concentration of 10-200 μg / mL in an oxygen-glucose deprivation / reperfusion (OGD / R) model to increase cell viability by 30%-50% and reduce the levels of creatine kinase isoenzyme (CK-MB), cardiac troponin T (cTnT), and 8-hydroxy-2'-deoxyguanosine (8-OHdG).

10. The application according to claim 8, characterized in that, The inhibition of myocardial inflammation includes: downregulating the protein expression levels of myeloid differentiation factor 88 (MyD88) and CCAAT / enhancer-binding protein β (CEBPB) in cardiomyocytes, and / or inhibiting the phosphorylation of nuclear factor κBp65 subunit (NF-κBp65) and p38 mitogen-activated protein kinase (p38MAPK).

11. The application according to any one of claims 8-10, characterized in that, The product is a drug, functional food, nutritional supplement or food additive, used to prevent or alleviate myocardial oxidative damage related to cardiovascular disease, wherein the amount of chicory aerial polysaccharide (CP-A) added is 0.1%-10.0% (w / v).

12. A myocardial cell protective composition, characterized in that, It contains an effective amount of the chicory aerial polysaccharide (CP-A) as described in claim 6 or 7 as the active ingredient, and a pharmaceutically or food-grade acceptable carrier or excipient.

13. The composition according to claim 12, characterized in that, Further comprising chicory root polysaccharide (CP-R), wherein the mass ratio of CP-A to CP-R is 1:20-20:1, wherein CP-R is prepared from chicory roots using the same extraction method as CP-A, and CP-A and CP-R are equivalent in cardiomyocyte protective activity, showing no significant difference in protective effect against OGD / R-induced H9c2 cardiomyocyte damage at the same concentration (p>0.05).

14. The composition according to claim 12 or 13, characterized in that, The composition is a pharmaceutical composition, health food, or functional food additive, used to synergistically exert antioxidant and anti-inflammatory effects.

15. An evaluation method for comparing the cardioprotective activity of polysaccharides from different parts of chicory, characterized in that, Includes the following steps: (i) Polysaccharides were extracted from the roots and aerial parts of chicory using the methods described in any one of claims 1-5 to obtain root polysaccharide (CP-R) and aerial polysaccharide (CP-A); (ii) An oxygen-glucose deprivation / reperfusion (OGD / R) injury model was established in H9c2 cardiomyocytes; (iii) Cells in the injury model were treated with CP-R and CP-A at concentrations of 10-200 μg / mL, respectively; (iv) The cardiomyocyte protective activities of CP-R and CP-A were evaluated and compared by detecting cell viability, myocardial injury markers (including CK-MB and cTnT), and oxidative stress markers (including 8-OHdG); (v) The protein expression levels of MyD88, CEBPB, phosphorylated NF-κBp65, and phosphorylated p38MAPK in cardiomyocytes were detected by Western blotting to elucidate the molecular mechanisms of CP-R and CP-A; wherein, if CP-A exhibited protective activity comparable to CP-R, the aerial parts of chicory were determined to be a substitute source of polysaccharides with cardiomyocyte protective activity.

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