CN118319987A - Application of papaya extract polyphenol in preparation of medicines for treating osteoarthritis - Google Patents
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/73—Rosaceae (Rose family), e.g. strawberry, chokeberry, blackberry, pear or firethorn
- A61K36/732—Chaenomeles, e.g. flowering quince
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P19/00—Drugs for skeletal disorders
- A61P19/02—Drugs for skeletal disorders for joint disorders, e.g. arthritis, arthrosis
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
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Abstract
The invention provides an application of wrinkled papaya polyphenol in preparing a medicament for treating osteoarthritis. Use of said wrinkled papaya polyphenol in the manufacture of a medicament for reducing the release of pro-inflammatory cytokines and inflammatory mediators NO in primary chondrocytes, and/or for reducing the mRNA expression of an inflammation-related enzyme, and/or for inhibiting the phosphorylation of NF- κb signaling pathway-related proteins. In order to test and clearly determine the anti-inflammatory effect of the papaya polyphenol, a ADTC primary chondrocyte inflammation model is constructed and observed. The results show that the wrinkled papaya polyphenol has remarkable inhibition effect on IL-1 beta induction ADTC primary chondrocyte inflammation, can effectively reduce secretion of pro-inflammatory cytokines (IL-6 and TNF-alpha), inhibit release of inflammatory mediators NO, down regulate mRNA expression of inflammatory enzymes (iNOS and Cox-2), and reduce phosphorylation of I kappa B alpha and p65 protein.
Description
Technical Field
The invention relates to the field of medicines, in particular to application of polyphenol extracted from chaenomeles speciosa in preparing medicines for treating osteoarthritis.
Background
Osteoarthritis (OA) is a chronic degenerative joint disease with complex etiology, which uses destruction and degeneration of articular cartilage as a main pathological feature, and joint pain and dyskinesia as main clinical symptoms, and is frequently found in joints such as knee, hip, hand and foot, etc., and is often found in obese elderly people, and is an important factor for endangering health of middle-aged and elderly people. The onset of the disease is mainly related to age, obesity, inheritance and other factors. The pathogenesis of the disease is not clear, and is mainly due to inflammatory injury of the articular cartilage cells, and the inflammatory injury is specifically represented by high secretion of proinflammatory cytokines and high expression of inflammatory enzymes in the cartilage cells, and high phosphorylation of NF- κB signal channel related proteins. Therefore, there is a sufficient reason to prove that the effect of reducing the inflammatory injury of articular cartilage thin and simultaneously playing the role of protecting cartilage cells is improved remarkably by the dual effects of the articular cartilage thin and inflammatory injuries.
The current common osteoarthritis treatment schemes in clinic comprise operation treatment and non-operation treatment, wherein the non-operation treatment mainly comprises drug treatment and aims at relieving symptoms, but no specific drug exists. Common drugs currently used to treat osteoarthritis include non-steroidal anti-inflammatory drugs (NSAIDS), hormonal drugs, and narcotic sedatives. Although the medicines are widely applied to clinic, the functions of organisms are affected due to the large toxic and side effects of long-term use, so that the medicines without toxic and side effects are required to be used for replacing classical medicines for treating clinical osteoarthritis.
Papaya belongs to plants of the genus papaya of the family Rosaceae, including wrinkled papaya, smooth papaya, chaenomeles sinensis and the like, is widely planted in China, is mainly distributed in China areas such as Hubei province, anhui province and the like, and has long planting history. The chaenomeles speciosa is also called as the dried fruit of chaenomeles speciosa, has fragrant and sour taste and slightly cold nature, contains various active ingredients such as polyphenol, flavone, polysaccharide, saponin, organic acid, terpenoid and the like, wherein the polyphenol is the most effective active ingredient in the chaenomeles speciosa, and the content of the polyphenol in the chaenomeles speciosa is obviously higher than that of other species, so the chaenomeles speciosa is commonly used as the best raw material for extracting the polyphenol of the chaenomeles speciosa. The wrinkled papaya enters four channels of spleen, lung, liver and kidney, and the polyphenol extracted from the wrinkled papaya has the effects of resolving dampness, resolving phlegm, easing pain, detumescence and relaxing tendons in traditional Chinese medicine. Meanwhile, the polyphenol extracted from the plant is used as a natural plant extract, so that the plant extract has the advantages of small toxic and side effects, less drug residues and high safety. To date, there has been no report on the prevention or treatment of osteoarthritis by polyphenols extracted from chaenomeles speciosa.
Disclosure of Invention
Aiming at the defects of the prior art, the invention provides application of the wrinkled papaya polyphenol in preparing a medicament for treating osteoarthritis, and the wrinkled papaya polyphenol has a good inhibiting effect on osteoarthritis and can be used for preparing the medicament for treating osteoarthritis.
The technical scheme of the invention is the application of the wrinkled papaya polyphenol in preparing the medicine for treating osteoarthritis.
The first object of the invention is to provide an application of wrinkled papaya polyphenol in preparing a medicine for treating osteoarthritis.
Use of said wrinkled papaya polyphenol in the manufacture of a medicament for reducing the release of pro-inflammatory cytokines and inflammatory mediators NO in primary chondrocytes, and/or for reducing the mRNA expression of an inflammation-related enzyme, and/or for inhibiting the phosphorylation of NF- κb signaling pathway-related proteins.
The pro-inflammatory cytokines are TNF-alpha and IL-1, the inflammation related enzymes are iNOS and Cox-2, and the NF- κB signal path related proteins are IκBalpha and p65.
The wrinkled papaya polyphenol is obtained by soaking dry wrinkled papaya powder in ethanol solution, extracting, drying and then performing enzymolysis by using cellulase.
The mass concentration of the ethanol solution is 95% or more.
The addition amount of the cellulase is 1.0-2.0% of the mass of the dry powder obtained by leaching the wrinkled papaya polyphenol with ethanol; the enzymolysis temperature is 50-60 ℃, the enzymolysis pH is 4.5-6.0, and the enzymolysis time is 2-4h.
A second object of the present invention is to provide a medicament for treating osteoarthritis, which comprises chaenomeles speciosa polyphenol as an active ingredient, and all pharmaceutically acceptable excipients or carriers.
Further, the pharmaceutical dosage form is often a suspension, capsule, tablet, pill, powder, injection, aerosol or other pharmaceutically acceptable dosage form. Generally, oral administration is usually solid preparations such as capsules, tablets, pills, powder and the like, or liquid preparations such as suspension, syrup and the like; the nasal inhalation is usually aerosol and other gas preparations; intravenous injection and intramuscular injection are often liquid preparations such as suspensions and injections.
The invention has the following benefits:
IL-1β, TNF- α and IL-6 are common inflammatory factors, whereas IL-1β has been reported to be highly expressed in animals and humans suffering from osteoarthritis. IL-1β stimulates high secretion of pro-inflammatory cytokines (TNF- α and IL-6) and inflammatory mediators NO in primary chondrocytes ADTC, and up-regulates mRNA expression of inflammatory enzymes (iNOS and Cox-2), and promotes phosphorylation of IκBα and p65 proteins, resulting in inflammatory injury of primary chondrocytes, thereby constructing an in vitro osteoarthritis model. Thus, we stimulated primary chondrocytes ADTC with IL-1 β to construct an in vitro model of osteoarthritis.
The current common osteoarthritis treatment schemes in clinic comprise operation treatment and non-operation treatment, wherein the non-operation treatment mainly comprises drug treatment and aims at relieving symptoms, but no specific drug exists. Common drugs currently used to treat osteoarthritis include non-steroidal anti-inflammatory drugs (NSAIDS), hormonal drugs, and narcotic sedatives. Although the medicines are widely applied to clinic, the functions of organisms are affected due to the large toxic and side effects of long-term use, so that the medicines without toxic and side effects are required to be used for replacing classical medicines for treating clinical osteoarthritis. Based on the current research, the papaya polyphenol as a novel drug which has NO toxic or side effect and NO drug residue after long-term use is proved to play a good anti-inflammatory effect by reducing the release of proinflammatory cytokines (IL-6 and TNF-alpha) and inflammatory mediators NO in ADTC primary chondrocytes, down regulating the mRNA expression of inflammatory enzymes (iNOS and Cox-2) and inhibiting the phosphorylation of I kappa B alpha and p65 proteins, thereby further relieving osteoarthritis symptoms.
To date, there has been no report on the prevention or treatment of osteoarthritis by polyphenols extracted from chaenomeles speciosa.
In order to test and clearly determine the anti-inflammatory effect of the papaya polyphenol, a ADTC primary chondrocyte inflammation model is constructed and observed. The results show that the wrinkled papaya polyphenol has remarkable inhibition effect on IL-1 beta induction ADTC primary chondrocyte inflammation, can effectively reduce secretion of pro-inflammatory cytokines (IL-6 and TNF-alpha), inhibit release of inflammatory mediators NO, down regulate mRNA expression of inflammatory enzymes (iNOS and Cox-2), and reduce phosphorylation of I kappa B alpha and p65 protein.
Drawings
FIG. 1 is a hydrogen spectrum of polyphenol extracted from Chaenomeles speciosa in example 1.
FIG. 2 is a graph showing the effect of the polyphenol extracted from Chaenomeles speciosa in example 2 on cell viability.
FIG. 3 is a graph showing the effect of different concentrations of IL-1β on NO release from ADTC cells in example 3.
FIG. 4 is a graph showing the effect of the papaya extract polyphenol on IL-1β -induced ADTC cells to release NO in example 4; wherein, # # # P < 0.0001 compared to the placebo group; p < 0.05, P < 0.0001 compared to model group.
FIG. 5 is a graph showing the effect of papaya extract polyphenol on IL-1β -induced ADTC cells to release TNF- α in example 5; wherein, # # # P < 0.0001 compared to the placebo group; p < 0.0001 compared to model group.
FIG. 6 is a graph showing the effect of papaya extract polyphenol on IL-1. Beta. Induced ADTC cells to release IL-6 in example 5; wherein, # # # P < 0.0001 compared to the placebo group; p < 0.005 and P < 0.0001 compared to model group.
FIG. 7 is a graph showing the effect of the polyphenol extracted from Chaenomeles speciosa in example 6 on IL-1β induction ADTC cell iNOS mRNA expression; wherein, # # # P < 0.0001 compared to the placebo group; p < 0.005 and P < 0.0001 compared to model group.
FIG. 8 is a graph showing the effect of the papaya extract polyphenol on IL-1β -induced ADTC cell Cox-2 mRNA expression in example 6; wherein, # # # P < 0.0001 compared to the placebo group; p <0.01 and P < 0.0001 compared to model group.
FIG. 9 is a graph showing the analysis of the effect of the polyphenol extracted from Chaenomeles speciosa in example 7 on IL-1β -induced ADTC cell phosphorylation of p65 (p-p 65) protein level; wherein, # # # P < 0.0001 compared to the placebo group; p < 0.005 and P < 0.0001 compared to model group.
FIG. 10 is a graph showing the analysis of the effect of the polyphenol extracted from Chaenomeles speciosa in example 7 on IL-1β -induced ADTC cell phosphorylation IκBα (p-IκBα) protein level; wherein, # # # P < 0.0001 compared to the placebo group; p < 0.01 and P < 0.0001 compared to model group.
Detailed Description
The present invention will be described in detail with reference to the following examples, which are given by way of illustration only and are not intended to be limiting.
Example 1 preparation and extraction of Chaenomeles speciosa polyphenols
Preparation of dried powder of wrinkled papaya: the dried papaya with wrinkled skin is dried in electrothermal blowing drying oven at 40 deg.c for 12 hr, crushed in crusher and sieved with 50 mesh sieve to obtain dry papaya powder.
Degreasing of wrinkled papaya dry powder: 50.00 g g of dried wrinkled papaya powder is taken, 95% ethanol is added according to the feed-liquid ratio of 1:10, the ethanol is removed by suction filtration after refluxing for 2: 2h, and 35.0g of defatted wrinkled papaya powder is obtained after drying.
Extracting the wrinkled papaya polyphenol by an enzymolysis method: weighing 5.0 g of the dry defatted wrinkled papaya powder, adding distilled water according to a feed-liquid ratio of 1:30, regulating the pH value to 5, adding 1.5% of cellulase, carrying out enzymolysis at 52 ℃ for 2.5 hours, then raising the temperature to 80 ℃ to inactivate enzymes, cooling and carrying out suction filtration to obtain filtrate, thus obtaining the wrinkled papaya polyphenol extract (and carrying out the test of the following example), wherein the content of the wrinkled papaya polyphenol obtained by the method is 150.48 mg/g, and the yield is 15.05%. As shown in fig. 1, the extracted polyphenol has the following chemical formula:
Formula 1.
Example 2CCK8 method the effect of polyphenols from Chaenomeles speciosa extract on ADTC cell activity was examined.
ADTC5 primary chondrocytes were cultured in 10% foetal calf serum DMEM high sugar medium, placed in 37 ℃ cell incubator containing 5% CO 2, and subjected to liquid exchange once daily, and passaged once every 1-2 days. Cells were seeded at 1×10 4 cells/well in 96-well plates, three wells were placed in each group, and ADTC primary chondrocytes were treated with 0, 5, 10, 20, 40, 50, 100 μg/mL of basal medium of the chaenomeles speciosa polyphenol extract. After culturing cells 24h, 10 μl of CCK-8 solution was added to the culture medium, and after culturing in a 37 ℃ cell incubator for 1 hour, absorbance was measured at 450: 450 nm with a microplate reader, respectively. Cell viability was calculated according to the following formula:
Cell viability (%) = (OD Experimental group -OD blank group /OD Control group -OD blank group ) ×100
After ADTC primary chondrocytes are treated for 24 hours by using polyphenols (structural formula is shown as formula 1) extracted from the wrinkled papaya with different concentrations, CCK8 detection results are shown as figure 2, and when the concentration of the polyphenols of the wrinkled papaya is lower than 40 mug/mL, the survival rate of the cells is above 95%, and obvious cytotoxicity is not seen; when the concentration is higher than 40 mug/mL, the cell survival rate is obviously reduced by about 20%, and the cell has stronger toxic effect on ADTC primary chondrocytes (figure 2). Thus, 5, 10, 20 μg/mL of the example 1 Chaenomeles speciosa extract polyphenol was selected for subsequent related experiments.
Example 3
Treatment of ADTC primary chondrocytes with IL-1 beta to model osteoarthritis lesions
After plating cells at 1X 10 4 cells/well in 96-well plates and culturing in 37℃cell culture incubator at 5% CO 2 for 24 hours, three duplicate wells were placed for each group, ADTC5 cells were treated with different concentrations of IL-1β (3,6,9,12,15,18 and 21 ng/ml), and the control group was added with an equal amount of basal medium. After 12h of incubation, NO release was measured using Griess kit, and the amounts of NO produced in each group were compared to determine the optimal induction concentration.
After ADTC primary chondrocytes were treated with different concentrations for 12 hours, NO release was detected, and as shown in fig. 3, when IL-1β concentration was lower than 12 ng/mL, NO release increased with increasing concentration; when the concentration of IL-1 beta is higher than 12 ng/mL, excessive IL-1 beta damages ADTC5 primary chondrocytes, so that the NO release amount of the primary chondrocytes is obviously reduced. Furthermore, 12 ng/mLIL-1β treatment significantly increased ADTC cell NO release compared to the control group without IL-1β treatment (fig. 3). Based on the experimental results, 12 ng/mL IL-1β is the optimal induction concentration of the ADTC cell inflammation injury model.
Example 4
Effect of Chaenomeles speciosa extract polyphenols on ADTC cells releasing NO
After seeding cells at 1×10 4 cells/well in 96-well plates and culturing in 37 ℃ cell incubator at 5% CO 2 for 24h, three duplicate wells were placed in each group, 5, 10, 20 μg/mL of chaenomeles speciosa extract polyphenol (12 ng/mL IL-1 β for each group) was added to the drug group, 12 ng/mL IL-1 β was added to the model group, and an equal amount of basal medium was added to the control group. After 12 hours of incubation, the supernatants were collected, and the amounts of NO released were measured using the Griess kit, and the amounts of NO produced in each group were compared.
After ADTC cells are treated for 24 hours by using papaya polyphenol (structural formula is shown as formula 1) with different concentrations, the NO release amount is detected, and the result is shown as figure 4, and compared with a blank control group, the NO release amount of a model group is obviously increased, so that the experimental inflammation injury modeling is successful; the NO release in the drug group showed a significantly decreasing trend compared to the model group, and gradually decreased as the concentration of the papaya polyphenol increased (fig. 4). The experimental results show that the wrinkled papaya polyphenol effectively inhibits IL-1 beta from inducing ADTC primary chondrocytes to release NO.
Example 5
Effect of Chaenomeles speciosa extract polyphenols on the release of proinflammatory cytokines by ADTC primary chondrocytes
After cells were seeded at 1×10 4 cells/well in 96-well plates and cultured at 5% CO 2 in a 37℃cell incubator for 24. 24 h, three duplicate wells were placed in each group, 5, 10, 20 μg/mL of the extract of Chaenomeles speciosa polyphenol extracted in example 1 was added to the drug group (12 ng/mL IL-1β was added to each group), 12 ng/mL IL-1β was added to the model group, and an equal amount of basal medium was added to the control group. After 12h of incubation, the supernatant was collected. The levels of TNF- α and IL-6 were determined according to the instructions of ELISA kits.
After ADTC primary chondrocytes were treated with different concentrations of chaenomeles speciosa polyphenol (formula 1) for 24 hours, the concentrations of pro-inflammatory cytokines were measured, and the results are shown in fig. 5 and 6, and compared with the blank control group, the levels of pro-inflammatory cytokines TNF- α and IL-6 secreted by ADTC cells in the model group were significantly increased, which also indicates that the experimental inflammatory injury model was successfully constructed; compared with the model group, the content of proinflammatory cytokines secreted by cells in the drug group ADTC is obviously reduced, wherein the inhibition rate of 20 mug/mL of wrinkled papaya polyphenol on the proinflammatory cytokine release is most obvious, and the optimal anti-inflammatory effect is shown (figures 5 and 6). Based on the experiment, the papaya polyphenol obtained by the invention can effectively inhibit IL-1 beta from inducing ADTC cells to secrete pro-inflammatory cytokines.
Example 6
Effect of Chaenomeles speciosa extract polyphenols on the expression of inflammation-related enzyme mRNA
After seeding cells at 1×10 4/well in 96-well plates and culturing 24.sup. 24h in 37℃cell incubator with 5% CO 2, three duplicate wells were placed in each group, 5, 10, 20 μg/mL of the Chaenomeles speciosa polyphenol extract (12 ng/mL of IL-1β was added to each group) was added to the drug group, 12 ng/mL of IL-1β was added to the model group, and an equal amount of basal medium was added to the control group. After 24h of incubation, the supernatant was discarded and the cells were collected by centrifugation at 10 min at 2000 r/min at 4 ℃. Total RNA was extracted using an RNA extraction kit, and cDNA was synthesized from the extracted RNA using a reverse transcription kit. The PCR reaction conditions of the system are as follows: 95. pre-denaturation at 30s at 95℃for 5s and annealing at 60℃for 30s for 40 cycles. The invention takes GAPDH as an internal reference and adopts a Ct comparison method: 2 -ΔΔCT to calculate the relative expression level of the mRNA of the inflammation-associated enzyme.
After ADTC cells are treated for 24 hours by using polyphenols (the structural formula is shown as formula 1) with different concentrations of the chaenomeles speciosa extract, the relative expression quantity of inflammatory enzyme mRNA is detected by an RT-qPCR method, and the results show that compared with a blank control group, the expression quantity of inflammatory enzymes iNOS and Cox-2 mRNA of ADTC5 cells in a model group are greatly increased, which shows that the inflammatory enzyme activity of ADTC cells is obviously increased under the stimulation of IL-1 beta, thereby promoting the development of inflammation, and fully confirming the successful construction of an inflammation injury model of the invention; compared with the model group, the expression level of inflammatory enzymes iNOS and Cox-2 mRNA in the cells of the drug group ADTC showed a clear down-regulation trend, wherein 20 mug/mL of the wrinkled papaya polyphenol had the most significant inhibition effect on the expression of inflammatory enzymes iNOS and Cox-2 mRNA, which also indicates that 20 mug/mL is probably the drug concentration with the most obvious anti-inflammatory effect (FIGS. 7 and 8). Based on the experimental results, the polyphenol extracted from the chaenomeles speciosa is effective in inhibiting the expression of inflammatory enzyme mRNA.
Example 7
Effect of Chaenomeles speciosa extract polyphenols on NF- κB signaling pathway-related proteins
After seeding cells at 1×10 4/well in 96-well plates and culturing 24.sup. 24h in 37℃cell incubator with 5% CO 2, three duplicate wells were placed in each group, 5, 10, 20 μg/mL of the Chaenomeles speciosa polyphenol extract (12 ng/mL of IL-1β was added to each group) was added to the drug group, 12 ng/mL of IL-1β was added to the model group, and an equal amount of basal medium was added to the control group. After 24h of incubation, the supernatant was discarded and the cells were collected by centrifugation at 5000 r/min for 10min at 4 ℃. Adding a proper volume of cell lysate into cells, lysing for 40min at 4 ℃, centrifuging for 20min at 4 ℃ at 10000 r/min, collecting supernatant, and quantitatively determining the total amount of extracted proteins by using a BCA protein kit. And then detecting the expression levels of the IκBα and p65 proteins and phosphorylated proteins by using a Western Blot method.
After ADTC cells were treated with polyphenols (structural formula shown in formula 1) at different concentrations for 24 hours, the Wetern Blot method was used to detect the NF- κb signaling pathway-related protein expression, and the results are shown in fig. 10, in which the phosphorylation levels of ikbα and p65 were increased by 4.4 and 5.1 times, respectively, in model group ADTC cells under IL-1β stimulation, compared with the blank group; the phosphorylation levels of iκbα and p65 were significantly reduced in drug group ADTC cells compared to the model group, with 20 μg/mL of wrinkled papaya polyphenol having the most pronounced inhibitory effect on the phosphorylation levels of iκbα and p65 (fig. 9 and 10), consistent with the optimal anti-inflammatory concentrations in the examples described above. Based on the experiment, the polyphenol extracted from the chaenomeles speciosa is effective in inhibiting the expression of NF- κB signal channel related protein.
In summary, the invention provides an application of a wrinkled papaya extract polyphenol in preparation of a medicine for treating osteoarthritis. The polyphenol extracted from the chaenomeles speciosa can effectively reduce the release of pro-inflammatory cytokines (TNF-alpha and IL-1) and inflammatory mediators (NO) in primary chondrocytes, reduce the mRNA expression of inflammation-related enzymes (iNOS and Cox-2), inhibit the phosphorylation of NF- κB signal channel-related proteins (IκBalpha and p 65), reduce the inflammatory injury of chondrocytes, protect chondrocytes and effectively prevent and treat osteoarthritis. The polyphenol extracted from the chaenomeles speciosa nakai has wide prospect in preparing the medicines for treating osteoarthritis, which are safe and have high activity.
The above examples are preferred embodiments of the present invention, but the above examples are not limited to the embodiments of the present invention, and any other reasonable implementation method without departing from the principles and spirit of the present invention is within the scope of the present invention.
Claims (7)
1. Application of Chaenomeles speciosa polyphenol in preparing medicine for treating osteoarthritis is provided.
2. The use according to claim 1, wherein the use of the chaenomeles speciosa polyphenols for the manufacture of a medicament for reducing the release of pro-inflammatory cytokines and inflammatory mediators NO in primary chondrocytes, and/or for reducing the mRNA expression of inflammation-related enzymes, and/or for inhibiting the phosphorylation of NF- κb signaling pathway-related proteins.
3. The use according to claim 2, wherein the pro-inflammatory cytokines are TNF- α and IL-1, the inflammation-associated enzymes iNOS and Cox-2, and the nf- κb signaling pathway-associated proteins iκbα and p65.
4. The use according to any one of claims 1-3, wherein the wrinkled papaya polyphenol is wrinkled papaya polyphenol obtained by soaking and extracting wrinkled papaya dry powder with ethanol solution, drying and then performing enzymolysis with cellulase.
5. The use according to claim 4, wherein the mass concentration of the ethanol solution is 95% or more.
6. The application of claim 4, wherein the addition amount of the cellulase is 1.0-2.0% of the dry powder mass obtained by leaching the wrinkled papaya polyphenol with ethanol; the enzymolysis temperature is 50-60 ℃, the enzymolysis pH is 4.5-6.0, and the enzymolysis time is 2-4h.
7. The use according to claim 1, wherein the concentration of the wrinkled papaya polyphenol is 5-20 μg/mL.
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