A phospholipid nanomaterial derived from adventitious root extract of jasmonate-induced methyl jasmonate, as well as its preparation method, application, and pharmaceutical composition

Abstract

The present invention provides a methyl jasmonate-induced phospholipid nanomaterial from an extract of the adventitious roots of the Chinese yew vine, as well as a preparation method, application, and pharmaceutical composition thereof, relating to the field of biomedical technology. The methyl jasmonate-induced phospholipid nanomaterial from an extract of the adventitious roots of the Chinese yew vine provided by the present invention comprises an extract of the adventitious roots of the Chinese yew vine induced by the methyl jasmonate and a phospholipid bonded to the surface of the extract of the adventitious roots of the Chinese yew vine induced by the methyl jasmonate. The present invention bonds the extract of the adventitious roots of the Chinese yew vine induced by the methyl jasmonate with the phospholipid, thereby greatly improving the stability, water solubility, bioavailability, and pharmacological activity of the extract of the adventitious roots of the Chinese yew vine induced by the methyl jasmonate, and having excellent anti-inflammatory and anti-tumor activities. The phospholipid has low toxicity, and the methyl jasmonate-induced phospholipid nanomaterial from the adventitious roots of the Chinese yew vine induced by the methyl jasmonate is highly safe. The methyl jasmonate-induced phospholipid nanomaterial from the adventitious roots of the Chinese yew vine induced by the methyl jasmonate is a nanoscale biomaterial that greatly improves the efficiency of drug delivery.

Description

Technical Field

[0001] The present invention relates to the technical field of biomedicine, and in particular to a methyl jasmonate-induced phospholipid nanomaterial extracted from the adventitious roots of Xanthoceras chinensis, a preparation method thereof, an application thereof, and a pharmaceutical composition thereof. Background Art

[0002] Dendropanax morbifera, a member of the Araliaceae family, is an evergreen tree found in tropical and subtropical regions, often as a small understory shrub. These plants have upright stems that can grow up to 5 meters tall; the leaves may be unlobed or have two, three, or five lobes. Members of this genus have a moderate growth rate, reaching up to 1.5 feet (about 45 centimeters) per year. The flowers are yellow-green, the bark smooth and pale yellow-gray, and the umbels can reach a diameter of up to 0.4 inches (about 1 centimeter). Members of this genus typically prefer well-drained soil and partial shade. Various parts of the plant, such as seeds, leaves, roots, and stems, have been used in traditional medicine to treat headaches, infectious diseases, skin problems, and other ailments and are registered with the Korean Ministry of Food and Drug Safety (KMFDS). Dendropanax morbifera contains polyphenols, essential oils, phenols, flavonoids, tannins, terpenes, and alkaloids. Previous studies have identified a variety of biological activities of yellow sumac, such as antioxidant, anti-inflammatory, memory-enhancing, neuroprotective, anticancer, antidiabetic, hepatoprotective, immunomodulatory, antimalarial, cytotoxic, and larvicidal properties.

[0003] The seeds of S. truncatum have low tolerance to cold stress. The harvest of seeds requires a flowering and fruiting period of more than 6 years. In addition, it was noted that seeds obtained through long-term breeding methods showed a relatively low germination rate. Due to the geographical limitations of the growing environment of S. truncatum, it is difficult to plant in other areas, and seed production takes a longer time. In vitro reproduction of S. truncatum species, especially research on compounds obtained from in vitro propagation materials Jasmonic acid (JA) and its derivative methyl jasmonate (MeJA) are plant hormones that are widely used in induction studies in in vitro culture systems. Studies have found that methyl jasmonate can effectively increase the content of 3,5-dicaffeoylquinic acid in the adventitious roots of S. truncatum, and the increase of this compound significantly enhances the anti-inflammatory and anti-lung cancer efficacy of the adventitious root extract of S. truncatum.

[0004] The adventitious roots of the yellow lacquer tree contain polyphenols, flavonoids, terpenes, and alkaloids. These compounds are physically and chemically fragile, insoluble in water, or have negative effects. Improving the stability, water solubility, and bioavailability of the adventitious root extract of the yellow lacquer tree has important application value. Summary of the Invention

[0005] In view of this, the object of the present invention is to provide a methyl jasmonate-induced phospholipid nanomaterial extracted from the adventitious roots of the Chinese yew tree, as well as its preparation method, application, and pharmaceutical composition. The methyl jasmonate-induced phospholipid nanomaterial extracted from the adventitious roots of the Chinese yew tree provided by the present invention has strong stability, good water solubility, and high bioavailability.

[0006] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0007] The invention provides a methyl jasmonate-induced scutellaria truncatum adventitious root extract phospholipid nanomaterial, comprising the methyl jasmonate-induced scutellaria truncatum adventitious root extract and a phospholipid bonded to the surface of the methyl jasmonate-induced scutellaria truncatum adventitious root extract.

[0008] Preferably, the mass ratio of the extract of the adventitious roots of the Rhododendron chinense induced by methyl jasmonate to phospholipid is 1:0.5-3.

[0009] Preferably, the phospholipid is a natural phospholipid and / or a synthetic phospholipid; the natural phospholipid includes one or more of soybean lecithin, egg yolk lecithin and phosphatidylcholine.

[0010] The present invention provides a method for preparing a phospholipid nanomaterial from the adventitious root extract of Xanthoceras chinensis induced by methyl jasmonate as described in the above technical solution, comprising the following steps:

[0011] The methyl jasmonate-induced yellow varnish wood adventitious root extract solution and the phospholipid solution are mixed, and the solvent is removed, dissolved in water and compounded in sequence to obtain the methyl jasmonate-induced yellow varnish wood adventitious root extract phospholipid nanomaterial.

[0012] Preferably, the solvent in the methyl jasmonate-induced yellow lacquer wood adventitious root extract solution includes one or more of tetrahydrofuran, dichloromethane, chloroform, n-hexane, ethanol, ethyl acetate, cyclohexane, methanol, isopropanol, n-butanol and acetone; the concentration of the methyl jasmonate-induced yellow lacquer wood adventitious root extract solution is 1 to 10 g / L.

[0013] Preferably, the solvent in the phospholipid solution includes one or more of dichloromethane, chloroform and ethyl acetate; and the concentration of the phospholipid solution is 1 to 10 g / L.

[0014] Preferably, the mixing temperature is 25-45°C and the mixing time is 1-30 minutes;

[0015] The compounding time is 1 to 2 hours, and the compounding is carried out under shaking conditions, and the shaking speed is 60 to 100 rpm.

[0016] Preferably, after the compounding, the method further comprises: subjecting the obtained composite system to ultrasonic treatment, water dialysis and drying in sequence.

[0017] The present invention provides the use of the phospholipid nanomaterial of the adventitious root extract of Xanthoceras chinensis induced by methyl jasmonate described in the above technical solution or the phospholipid nanomaterial of the adventitious root extract of Xanthoceras chinensis induced by methyl jasmonate prepared by the preparation method described in the above technical solution in preparing medicines.

[0018] The present invention provides a pharmaceutical composition comprising an active component and a pharmaceutically acceptable excipient, wherein the active component comprises the phospholipid nanomaterial of the adventitious root extract of Xanthoceras chinensis induced by methyl jasmonate as described in the above technical solution or the phospholipid nanomaterial of the adventitious root extract of Xanthoceras chinensis induced by methyl jasmonate as described in the above technical solution.

[0019] The invention provides a methyl jasmonate-induced jasmine wood adventitious root extract phospholipid nanomaterial (MeJA-DMARE-Phytosome), comprising methyl jasmonate-induced jasmine wood adventitious root extract (MeJA-DMARE) and phospholipid bonded to the surface of the methyl jasmonate-induced jasmine wood adventitious root extract.

[0020] Phospholipids have amphiphilic properties, that is, they have both a hydrophilic head and a hydrophobic tail. The present invention embeds the methyl jasmonate-induced yellow lacquer wood adventitious root extract in phospholipids, which can form a lipid-compatible molecular structure in a non-polar solvent, greatly improving the water solubility of the methyl jasmonate-induced yellow lacquer wood adventitious root extract. Phospholipids not only serve as carriers, but also have multiple therapeutic properties. When applied to a drug system in which the methyl jasmonate-induced yellow lacquer wood adventitious root extract is an active ingredient, a double-product effect can be produced, which improves the biological activity, bioavailability and pharmacological activity of the methyl jasmonate-induced yellow lacquer wood adventitious root extract. The anti-inflammatory and anti-tumor activities of the phospholipid nanomaterials of the methyl jasmonate-induced yellow lacquer wood adventitious root extract are high. The lipid-based nanoparticle system of the methyl jasmonate-induced yellow lacquer wood adventitious root extract phospholipid nanomaterial provided by the present invention, as one of the most promising colloidal carriers of bioactive organic compounds, has excellent time stability (i.e., storage stability) and thermal stability, high loading capacity, and greatly improves the efficiency of drug delivery. Furthermore, the methyl jasmonate-induced extract of the yellow lacquerwood adventitious root extract and the polar phosphatidyl groups are not simply physically mixed, but rather hydrogen-bonded, significantly enhancing the stability of the methyl jasmonate-induced yellow lacquerwood adventitious root extract phospholipid nanomaterial. Furthermore, phospholipids have low toxicity, making the methyl jasmonate-induced yellow lacquerwood adventitious root extract phospholipid nanomaterial highly safe.

[0021] The invention provides a method for preparing a phospholipid nanomaterial from an adventitious root extract of Psoralea corylifolia induced by methyl jasmonate, which has simple process, simple operation, easy preparation, low manufacturing cost, high efficiency, and is easy to realize large-scale industrial production. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 This is the result of Tyndall effect of phospholipid nanomaterials aqueous solution extracted from adventitious roots of Xanthoceras chinensis induced by methyl jasmonate;

[0023] Figure 2 Scanning electron microscopy (A and B) and transmission electron microscopy (C and D) images of phospholipid nanomaterials extracted from the adventitious roots of Psoralea corylifolia induced by methyl jasmonate;

[0024] Figure 3 The dynamic light scattering analysis results of phospholipid nanomaterials extracted from the adventitious roots of Xanthoceras chinensis induced by methyl jasmonate, where A is the particle size distribution diagram and B is the zeta potential diagram;

[0025] Figure 4 UV-visible spectra of the extract of the adventitious roots of the yellow lacquer tree induced by methyl jasmonate, the phospholipid nanomaterials and phosphatidylcholine of the extract of the adventitious roots of the yellow lacquer tree induced by methyl jasmonate;

[0026] Figure 5 The infrared spectra of the extract of the adventitious root of the yellow lacquer tree induced by methyl jasmonate, the phospholipid nanomaterials and phosphatidylcholine of the extract of the adventitious root of the yellow lacquer tree induced by methyl jasmonate;

[0027] Figure 6 Figure 2 shows the inhibitory effects of methyl jasmonate-induced adventitious root extracts of Echeveria serrata and methyl jasmonate-induced phospholipid nanomaterials of Echeveria serrata adventitious root extracts on NO production in RAW cells;

[0028] Figure 7 Figure 2 shows the inhibitory effects of methyl jasmonate-induced adventitious root extracts of Xanthoceras chinensis and methyl jasmonate-induced phospholipid nanomaterials of Xanthoceras chinensis adventitious root extracts on ROS generation in RAW cells;

[0029] Figure 8 These are graphs showing the inhibitory expression results of the extract of the adventitious roots of Echinops chinensis induced by methyl jasmonate and the phospholipid nanomaterials of the adventitious root extract of Echinops chinensis induced by methyl jasmonate on the levels of inflammation-related cytokines, wherein A is an agarose gel electrophoresis band diagram of the mRNA gene, B is an mRNA expression diagram of the IL-6 gene, C is an mRNA expression diagram of the iNOS gene, D is an mRNA expression diagram of the TNF-α gene, E is an mRNA expression diagram of the COX-2 gene, and F is an mRNA expression diagram of the IL-1β gene;

[0030] Figure 9 This figure shows the results of the promotion effect of methyl jasmonate-induced adventitious root extract of Xanthoceras chinensis and methyl jasmonate-induced phospholipid nanomaterials of Xanthoceras chinensis adventitious root extract on ROS generation in A549 lung cancer cells;

[0031] Figure 10 The graphs show the inhibitory effects of methyl jasmonate-induced extracts of the adventitious roots of A. chinensis and methyl jasmonate-induced phospholipid nanomaterials of the adventitious root extracts of A. chinensis on the migration of A549 lung cancer cells, wherein A is a graph showing the migration of A549 lung cancer cells, and B is a graph showing the migration amount of A549 lung cancer cells;

[0032] Figure 11 The figures show the regulatory effects of methyl jasmonate-induced extracts of the adventitious roots of Echinops chinensis and methyl jasmonate-induced phospholipid nanomaterials of the adventitious root extracts of Echinops chinensis on the expression of apoptotic genes in A549 lung cancer cells, wherein A and F are agarose gel electrophoresis bands of mRNA genes (from left to right: control, MeJA-DMARE, and MeJA-DMARE-phytosome), B is the mRNA expression level of the Caspase 3 gene, C is the mRNA expression level of the Bax gene, D is the mRNA expression level of the Bcl-2 gene, E is the mRNA expression level of the Caspase 9 gene, G is the mRNA expression level of the p38 gene, H is the mRNA expression level of the JNK gene, I is the mRNA expression level of the Nrf2 gene, J is the mRNA expression level of the HO-1 gene, and K is the mRNA expression level of the CAT gene. DETAILED DESCRIPTION

[0033] The invention provides a methyl jasmonate-induced jasmine adventitious root extract phospholipid nanomaterial, comprising the methyl jasmonate-induced jasmine adventitious root extract (denoted as MeJA-DMARE) and a phospholipid bonded to the surface of the methyl jasmonate-induced jasmine adventitious root extract.

[0034] In the present invention, the mass ratio of the methyl jasmonate-induced Psoralea corylifolia adventitious root extract to phospholipids is preferably 1:0.5-3, more preferably 1:1-2.5, further preferably 1:1.5-2, and particularly preferably 1:1, 1:2 or 1:3.

[0035] In the present invention, the phospholipid is preferably a natural phospholipid and / or a synthetic phospholipid; the natural phospholipid preferably includes one or more of soybean lecithin, egg yolk lecithin, and phosphatidylcholine, more preferably phosphatidylcholine. In the present invention, the synthetic phospholipid preferably includes one or more of soybean lecithin, egg yolk lecithin, and phosphatidylcholine, more preferably phosphatidylcholine.

[0036] In the present invention, the bonding is preferably through hydrogen bonding.

[0037] The present invention provides a preparation method of a phospholipid nanomaterial from the adventitious root extract of Xanthoceras chinensis induced by methyl jasmonate according to the above technical solution, comprising the following steps: mixing a solution of the adventitious root extract of Xanthoceras chinensis induced by methyl jasmonate and a phospholipid solution, and sequentially performing solvent removal, water dissolution, and compounding to obtain the phospholipid nanomaterial from the adventitious root extract of Xanthoceras chinensis induced by methyl jasmonate.

[0038] Unless otherwise specified, the materials and equipment used in the present invention are all commercially available products in the art.

[0039] In the present invention, the solvent in the methyl jasmonate-induced yellow lacquerwood adventitious root extract solution preferably includes one or more of tetrahydrofuran, dichloromethane, chloroform, n-hexane, ethanol, ethyl acetate, cyclohexane, methanol, isopropanol, n-butanol and acetone, more preferably methanol or ethanol. In the present invention, the concentration of the methyl jasmonate-induced yellow lacquerwood adventitious root extract solution is preferably 1-10 g / L, more preferably 2-6 g / L, and even more preferably 2-5 g / L. In the present invention, the methyl jasmonate-induced yellow lacquerwood adventitious root extract solution is preferably obtained by dissolving the methyl jasmonate-induced yellow lacquerwood adventitious root extract in a solvent; the dissolution temperature is preferably room temperature; the dissolution is preferably carried out under stirring conditions. The present invention has no special limitation on the stirring time, which is based on obtaining a clear and transparent liquid; specifically, stirring for 1-30 minutes, more preferably 5-10 minutes.

[0040] In the present invention, the method for preparing the extract of the adventitious roots of the yellow lacquer tree induced by methyl jasmonate preferably comprises the following steps: mixing the adventitious root powder of the yellow lacquer tree induced by methyl jasmonate with an extractant, performing extraction, concentrating the resulting extract, and then drying to obtain the adventitious root extract of the yellow lacquer tree induced by methyl jasmonate. In the present invention, the adventitious root powder of the yellow lacquer tree induced by methyl jasmonate is preferably obtained by drying, grinding, and sieving the adventitious roots of the yellow lacquer tree induced by methyl jasmonate in sequence; the sieving is preferably through a 100-mesh sieve, and the portion below the sieve is the adventitious root powder of the yellow lacquer tree induced by methyl jasmonate. In the present invention, the methyl jasmonate-induced Dendropanax morbifera adventitious roots are preferably prepared according to the document "Xu, F.; Valappil, AK; Zheng, S.; Zheng, B.; Yang, D.; Wang, Q. 3,5-DCQA as a Major Molecule in MeJA-Treated Dendropanax morbifera Adventitious Root to Promote Anti-Lung Cancer and Anti-Inflammatory Activities. Biomolecules 2024, 14, 705." In the present invention, the solid-to-liquid ratio of the methyl jasmonate-induced Dendropanax morbifera adventitious root powder to the extractant is preferably 1 g:20-40 mL, more preferably 1 g:30-40 mL; the extractant preferably comprises an ethanol aqueous solution, and the volume fraction of ethanol in the ethanol aqueous solution is preferably 30-80%, more preferably 60-70%. In the present invention, the extraction temperature is preferably 40-80°C, more preferably 60-80°C; the number of extractions is preferably 1-5 times, more preferably 2-4 times, and even more preferably 3 times; the time for a single extraction is preferably 1-3 hours, more preferably 1-2 hours. In the present invention, the concentration is preferably evaporation, the evaporation temperature is preferably 30-60°C, more preferably 40-50°C; the evaporation pressure is preferably 10-60 MPa, more preferably 20-30 MPa. The present invention does not particularly limit the drying temperature and time, as long as the solvent can be completely removed.

[0041] In the present invention, the solvent in the phospholipid solution preferably includes one or more of dichloromethane, chloroform, and ethyl acetate, more preferably chloroform. In the present invention, the concentration of the phospholipid solution is preferably 1 to 10 g / L, more preferably 2 to 6 g / L, and specifically preferably 2 g / L, 4 g / L, or 6 g / L. In the present invention, the phospholipid solution is preferably obtained by dissolving phospholipids in a solvent; the dissolution temperature is preferably room temperature; the dissolution is preferably carried out under stirring conditions; the present invention has no particular limitation on the stirring time, which is based on obtaining a clear and transparent liquid; specifically, 1 to 30 minutes, more preferably 5 to 10 minutes.

[0042] In the present invention, the mass ratio of the methyl jasmonate-induced yellow lacquer wood adventitious root extract in the methyl jasmonate-induced yellow lacquer wood adventitious root extract solution to the phospholipids in the phospholipid solution is preferably 1:0.5-3, more preferably 1:1-2.5, further preferably 1:1.5-2, and specifically preferably 1:1, 1:2 or 1:3.

[0043] In the present invention, the mixing temperature is preferably 25 to 45° C., more preferably 30 to 35° C.; the mixing time is preferably 1 to 30 min, more preferably 5 to 10 min; and the mixing is preferably performed under stirring conditions.

[0044] In the present invention, the desolventizing preferably includes rotary evaporation desolventizing, and the temperature of the desolventizing is preferably 30-60°C, more preferably 35-45°C, specifically preferably 30°C, 35°C, 36°C, 40°C, 45°C, 50°C, 55°C or 60°C; the present invention has no special limitation on the time of the desolventizing, and is based on the formation of a layer of attachment film visible to the naked eye (the solvent is completely removed).

[0045] In the present invention, the water for water dissolution preferably includes deionized water; the water dissolution is preferably performed under ultrasonic conditions, preferably at room temperature; the present invention has no particular limitation on the duration of the ultrasonication, and the ultrasonication can be performed until the attached film is completely dissolved. The present invention has no particular limitation on the amount of water used, and any amount sufficient to dissolve the methyl jasmonate-induced adventitious root extract of the yellow lacquer tree, specifically 1 to 10 g / L.

[0046] In the present invention, the temperature of the compounding is preferably room temperature, the time of the compounding is preferably 1 to 2 hours, more preferably 1 to 1.5 hours; the compounding is preferably carried out under shaking conditions, the shaking speed is preferably 60 to 100 rpm, more preferably 70 to 80 rpm; the shaking is preferably carried out on a shaker.

[0047] After completing the compounding, the present invention preferably further comprises: subjecting the obtained composite system to ultrasonic treatment, water dialysis and drying in sequence to obtain the methyl jasmonate-induced phospholipid nanomaterial of the adventitious root extract of the yellow lacquer wood. In the present invention, the temperature of the ultrasonic treatment is preferably room temperature, and the time is preferably 10 to 60 minutes, more preferably 20 to 30 minutes; the purpose of the ultrasonic treatment is to disperse the complex evenly. In the present invention, the molecular weight cutoff of the water dialysis is preferably 6000 to 8000 Da, more preferably 6500 to 7500 Da, and further preferably 7000 Da; the time of the water dialysis is preferably 1 to 3 hours, more preferably 2 hours. In the present invention, the drying preferably includes spray drying or freeze drying, more preferably freeze drying; the present invention has no special restrictions on the conditions of the drying, as long as the solvent can be completely removed.

[0048] The present invention provides the use of the methyl jasmonate-induced phospholipid nanomaterial of the adventitious root extract of the yellow lacquer tree described in the above technical solution or the methyl jasmonate-induced phospholipid nanomaterial of the adventitious root extract of the yellow lacquer tree obtained by the preparation method described in the above technical solution in the preparation of medicines. In the present invention, the medicine preferably includes an anti-inflammatory drug or an anti-tumor drug; the tumor preferably includes lung cancer. In the methyl jasmonate-induced phospholipid nanomaterial of the adventitious root extract of the yellow lacquer tree provided by the present invention, the methyl jasmonate-induced phospholipid nanomaterial of the adventitious root extract of the yellow lacquer tree is wrapped by phospholipids, which greatly improves the solubility of the methyl jasmonate-induced phospholipid nanomaterial of the adventitious root extract of the yellow lacquer tree, promotes its bioavailability, and has good anti-inflammatory and anti-cancer effects. Moreover, the methyl jasmonate-induced phospholipid nanomaterial of the adventitious root extract of the yellow lacquer tree provided by the present invention is a nano-scale biomaterial, which greatly improves the efficiency of drug delivery.

[0049] The present invention also provides a pharmaceutical composition comprising an active ingredient and a pharmaceutically acceptable excipient. The active ingredient is the methyl jasmonate-induced phospholipid nanomaterial from the adventitious root extract of the jasmine vine described in the above technical solution, or the methyl jasmonate-induced phospholipid nanomaterial from the adventitious root extract of the jasmine vine obtained by the preparation method described in the above technical solution. The present invention does not specifically limit the pharmaceutically acceptable excipients; pharmaceutically acceptable excipients familiar to those skilled in the art may be used. The present invention does not specifically limit the content of the active ingredient in the pharmaceutical composition, the pharmaceutical dosage form, or the preparation method of the pharmaceutical composition; the content of the active ingredient in the pharmaceutical composition, the pharmaceutical dosage form, and the preparation method of the pharmaceutical composition known to those skilled in the art may be used.

[0050] To further illustrate the present invention, the following detailed description of a methyl jasmonate-induced phospholipid nanomaterial from the adventitious roots of Psoralea corylifolia, its preparation method, application, and pharmaceutical composition is provided by the present invention in conjunction with the examples. However, these examples should not be construed as limiting the scope of protection of the present invention.

[0051] Example 1

[0052] The harvested adventitious roots of M. jasmonate-induced E. chinensis were dried, ground, and passed through a 100-mesh sieve. The sieved portion was the M. jasmonate-induced E. chinensis adventitious root powder. The M. jasmonate-induced E. chinensis adventitious root powder was mixed with a 70 vol% ethanol aqueous solution and ultrasonically extracted at 80°C for 1 hour. The supernatant was collected after each centrifugation. This extraction was repeated three times. The combined supernatants were distilled and dried to a constant weight at 10-60 MPa and 30-60°C to obtain the M. jasmonate-induced E. chinensis adventitious root extract (MeJA-DMARE). The solid-to-liquid ratio of the M. jasmonate-induced E. chinensis adventitious root powder to the 70 vol% ethanol aqueous solution used for the single extraction was 1 g:40 mL.

[0053] Methyl jasmonate-induced adventitious root extract of Ficus microcarpa was magnetically stirred with methanol until clear and transparent, yielding a 10 mg / mL MeJA-DMARE solution. Phosphatidylcholine (PC) was stirred with chloroform until clear and transparent, yielding a 2 mg / mL PC solution. The MeJA-DMARE and PC solutions were magnetically stirred at 30°C for 10 minutes, and the solvent was evaporated to dryness at 60 MPa and 45°C. Deionized water was added and sonicated until a thin film was completely dissolved in the deionized water, yielding a mixed aqueous solution (MeJA-DMARE concentration of 2 mg / mL). The mixed aqueous solution was shaken on a shaker at room temperature (25°C) and 70 rpm for 1 hour, followed by sonication in a water bath for 20 minutes and dialyzed for 2 hours using a 6000-8000 Da dialysis membrane. The dialyzed solution was freeze-dried to constant weight to yield the methyl jasmonate-induced phospholipid nanomaterial (MeJA-DMARE-Phytosome complex) derived from the adventitious root extract of Ficus microcarpa. The mass ratio of MeJA-DMARE to PC is 1:1.

[0054] The MeJA-DMARE-Phytosome prepared in this example had an encapsulation efficiency of 43.14±2.86% for MeJA-DMARE and a drug loading capacity of 36.50±3.64%. The encapsulation efficiency and drug loading of the complex were determined by ultracentrifugation. The specific steps are as follows: 1 mg of MeJA-DMARE-Phytosome was centrifuged at 4°C and 15,000 rpm for 90 minutes using a refrigerated centrifuge. The supernatant λ was measured using a UV-visible spectrophotometer. max = absorbance at 310 nm and use the calibration curve (concentration range 0.05-1 mg / mL, R 2=0.998 (n=3)) to determine the amount of MeJA-DMARE, and the linear regression equation was y = 3.2607x + 0.0305. Entrapment efficiency = (actual measured mass content of MeJA-DMARE in MeJA-DMARE-Phytosom / theoretical mass content of MeJA-DMARE in MeJA-DMARE-Phytosom) × 100%. Drug loading = (actual measured mass of MeJA-DMARE in MeJA-DMARE-Phytosom / mass of MeJA-DMARE-Phytosom) × 100%.

[0055] Example 2

[0056] Methyl jasmonate-induced phospholipid nanomaterials derived from the adventitious roots of Eupatorium truncatum were prepared according to the method of Example 1. The only differences from Example 1 were that the MeJA-DMARE solution contained ethanol, the PC solution concentration was 6 mg / mL, and the mass ratio of MeJA-DMARE to PC was 1:3. The prepared MeJA-DMARE-Phytosome exhibited a MeJA-DMARE encapsulation efficiency of 57.90±1.87% and a drug loading of 54.98±4.90%.

[0057] Example 3

[0058] Methyl jasmonate-induced phospholipid nanomaterials derived from the adventitious roots of Psoralea corylifolia were prepared according to the method of Example 1. The only differences from Example 1 were that the PC solution concentration was 4 mg / mL and the mass ratio of MeJA-DMARE to PC was 1:2. The prepared MeJA-DMARE-Phytosome exhibited an encapsulation efficiency of 79.98±1.45% for MeJA-DMARE and a drug loading of 69.17±0.14%.

[0059] Test Example 1

[0060] (1) Tyndall effect

[0061] Tyndall effect is a phenomenon that often occurs in colloidal solutions. The prepared complex was tested by Tyndall effect. The Tyndall effect can be used to infer the dispersion of MeJA-DMARE-Phytosome. The results are shown in Figure 1 It can be seen that the bright light beam can penetrate the MeJA-DMARE-Phytosome aqueous solution visually, which preliminarily determines that MeJA-DMARE-Phytosome is a colloid with uniformly distributed particles.

[0062] (2) Scanning electron microscope

[0063] The shape, morphology and distribution of MeJA-DMARE-Phytosome were determined by field emission transmission microscopy (FE-TEM) and field emission scanning electron microscopy (FE-SEM). Figure 2 A and B in the middle, transmission electron microscopy images are shown in Figure 2 C and D. Scanning electron microscopy images show that the liposome-like vesicles exhibit a contiguous structure, uniform size, and good dispersion. Transmission electron scanning images reveal that the MeJA-DMARE-Phytosomes are spherical and translucent, self-enclosed, with a rough surface and no signs of particle aggregation. Clearly, MeJA-DMARE (MeJA-DMARE) forms a spherical structure when bound to phosphatidylcholine (PC).

[0064] (3) Dynamic light scattering results analysis

[0065] The size distribution, particle dispersion coefficient and zeta potential difference of MeJA-DMARE-Phytosome in water were evaluated using an ELS-Z2 series particle size analyzer at 25°C. Figure 3 , where A is the particle size distribution diagram and B is the zeta potential diagram. The results show that the prepared MeJA-DMARE-Phytosome has an average particle size of 210 nm, a small particle size, uniform size distribution, a small dispersion coefficient (0.16), and an excellent zeta potential (-25.19 mV).

[0066] (4) UV spectrum scanning

[0067] The UV absorption spectrum can reflect the changes in the unsaturated bonds of the compound. MeJA-DMARE, MeJA-DMARE-Phytosome and phosphatidylcholine were made into ethanol solutions of the same concentration and scanned by UV-visible spectroscopy. The results are shown in Figure 4 It can be seen that phosphatidylcholine has a characteristic absorption peak at 235nm, MeJA-DMARE has a maximum absorption peak at 310nm, and MeJA-DMARE-Phytosome also has a maximum absorption peak at this wavelength, but the peak value is not as high as that of MeJA-DMARE of equal concentration. This proves that MeJA-DMARE-Phytosome has not undergone any change in its chemical structure and mainly exhibits the chemical characteristics of MeJA-DMARE. However, because the characteristic structure may be involved in the binding of MeJA-DMARE to phospholipids, the peak value of the characteristic peak has decreased.

[0068] (5) Infrared spectrum scanning

[0069] Infrared spectroscopy is an absorption spectrum used to study molecular motion. The position and shape of the absorption peaks in the infrared spectrum can be used to infer the functional group information of the compound. MeJA-DMARE-Phytosome was further characterized using Fourier transform infrared (FTIR) spectroscopy to study the interactions between the functional groups on MeJA-DMARE-Phytosome. For FTIR characterization, the dried plant extract, MeJA-DMARE-Phytosome, and PC powder were concentrated at 4000-450 cm -1 Scanning within the range with a resolution of 4cm -1 , the results are shown in Figure 5 It can be seen that the fingerprint area (below 1500cm -1 ) showed that a new plant MeJA-DMARE-Phytosome was generated. The spectrum at 3439 cm -1 The spectrum shows a peak at 2932 cm, indicating the presence of aliphatic alcohol (-OH) groups commonly found in polyphenolic compounds. -1 and 2957cm -1 The presence of long chain fatty acid bands further indicates the formation of phospholipids. -1 ) and POC(1055cm -1 ) corresponds to the peak of the phospholipid bond at the phospholipid bond P=O (1463cm -1 ) and phospholipid bond POC (1055cm -1 ) are shifted to 1469cm -1 and 1074cm -1 Unlike plant extracts, the MeJA-DMARE-Phytosome spectrum showed a synergistic effect on its components. For example, with the OH (3439 cm -1 ) and C=O (1644 cm -1 ) are related to the characteristic signals at 3476 cm in phospholipids. -1 and 1652cm -1 The peaks merged into a new peak at 100 nm. This suggests that hydrogen bonds formed between the OH groups of the plant extract and the polar groups of the phospholipids. Therefore, these relatively weak intermolecular interactions may be a contributing factor to the formation of MeJA-DMARE-Phytosome.

[0070] Test Example 2

[0071] Inhibitory effects of MeJA-DMARE, PC and MeJA-DMARE-Phytosome on NO production in RAW cells.

[0072] NO plays a role in host defense within the bronchial epithelium and acts as an inflammatory mediator in pathological conditions. Nitric oxide is produced in abnormal conditions, including inflammatory bowel disease, arthritis, osteoporosis, and various inflammatory respiratory diseases. Therefore, inhibiting the overproduction of NO has become an important target for the treatment of inflammatory diseases.

[0073] Cells were stimulated with 1 μg / mL LPS and then incubated for 24 h. RAW 264.7 cells were then pretreated with different concentrations of MeJA-DMARE and MeJA-DMARE-Phytosome for 1 h. Nitrite levels in the culture medium were assessed using Grise reagent. Briefly, 100 μL of Griess reagent was combined with 100 μL of supernatant. The absorbance at 540 nm was measured. L-NMMA was used as a positive control (standard inhibitor) in this experiment at a dose of 50 μM. Each experiment was repeated three times, and the results are shown in Figure 2. Figure 6 .

[0074] Compared to the control group, NO production was significantly increased in LPS-treated cells. However, NO production in cells induced with MeJA-DMARE and MeJA-DMARE-Phytosome was significantly reduced, even at the lowest concentration tested (31.25 μg / mL), by 40.5% and 38.9%, respectively, compared to LPS-treated cells. In each treatment group, the synthesized MeJA-DMARE-Phytosome exhibited a stronger ability to inhibit NO production than the plant extract.

[0075] Test Example 3

[0076] Inhibitory effects of MeJA-DMARE and MeJA-DMARE-Phytosome on ROS generation in RAW cells.

[0077] The cells were stimulated with 1 μg / mL LPS and then incubated for 24 hours. After that, RAW 264.7 cells were pretreated with different concentrations of MeJA-DMARE and MeJA-DMARE-Phytosome for 1 hour. After 24 hours of treatment in serum-free medium, the cells were washed three times with PBS and then exposed to 20 μM 2',7'-dichloro-dihydrofluorescein diacetate (DCFH-DA) at 37°C in the dark. After that, the medium was removed and the cells were washed twice with 100 μL PBS. The fluorescence intensity of ROS generation was detected. The results are shown in Figure 7 .

[0078] The results showed that cells exposed to LPS generated significantly higher amounts of ROS. In contrast, when LPS-induced cells were treated with MeJA-DMARE and MeJA-DMARE-phytosome, ROS production decreased in a dose-dependent manner. ROS production was significantly reduced at MeJA-DMARE concentrations exceeding 125 μg / mL. In each treatment group, the synthesized MeJA-DMARE-Phytosome demonstrated superior inhibition of LPS-induced ROS production in RAW 264.7 cells compared to the plant extract.

[0079] Test Example 4

[0080] MeJA-DMARE and MeJA-DMARE-Phytosome inhibit the expression of inflammation-related cytokine levels.

[0081] LPS-induced gene expression is controlled by a series of signaling pathways, including nuclear factor NF-κB, mitogen-activated protein kinases (MAPKs), and signal transducers and activators of transcription (STATs). NF-κB plays an important role in the development of inflammatory diseases and in regulating the transcription of inflammatory mediators such as iNOS, COX-2, TNF-α, and IL-1β. Macrophages highly express inducible NOS (iNOS), which is responsible for the synthesis of NO. The effects of MeJA-DMARE-phytosome on the expression of these factors were investigated using RT-PCR and qRT-PCR. Figure 8 Figure 1 shows the mRNA expression patterns of LPS-treated cells. Figures A and B show the mRNA expression of IL-6, C show the mRNA expression of iNOS, D show the mRNA expression of TNF-α, E show the mRNA expression of COX-2, and F show the mRNA expression of IL-1β. The results show that LPS-treated cells significantly increased the mRNA expression of COX-2, TNF-α, iNOS, IL-6, and IL-1β (8.86-fold, 7.38-fold, 11.67-fold, 8.70-fold, and 5.04-fold, respectively, compared to the untreated group). However, treatment with 250 μg / mL of MeJA-DMARE-phytosome reduced the mRNA expression of COX-2, TNF-α, iNOS, IL-6, and IL-1β by 1.16-fold, 2.31-fold, 2.37-fold, 0.11-fold, and 2.08-fold, respectively, compared to untreated cells.

[0082] From the comprehensive results of Test Examples 2 to 4, it can be seen that MeJA-DMARE-Phytosome has an anti-inflammatory effect.

[0083] Test Example 5

[0084] MeJA-DMARE and MeJA-DMARE-Phytosome promote ROS generation in A549 lung cancer cells.

[0085] The ROS content in A549 cells was measured using DCFH-DA. A594 cells were incubated at 37°C in a 5% CO2 environment for 24 hours. The harvested cells were treated with 125 μg / mL MeJA-DMARE and MeJA-DMARE-phytosome for 24 hours, and the ROS production was measured. Figure 9 As shown. The results showed that a dose-dependent increase in intracellular ROS generation was observed in A549 cells after treatment with 250 μg / mL and 500 μg / mL MeJA-DMARE-phytosome compared to the control group. Bioactive compounds can trigger apoptosis of cancer cells by promoting the accumulation of ROS, and MeJA-DMARE-phytosome can promote the generation of ROS. Previous studies have shown that in lung cancer cell lines, MeJA-DMARE-Phytosome has a stronger anti-tumor effect than free polyphenols. The MeJA-DMARE-Phytosome synthesized in the present invention can induce the production of ROS in cancer cells, which may trigger multiple cell death pathways and ultimately inhibit the progression of cancer.

[0086] Test Example 6

[0087] Inhibitory effects of MeJA-DMARE and MeJA-DMARE-Phytosome on the migration of A549 lung cancer cells.

[0088] The collective movement of cells in a cohesive manner is a significant feature of tissue remodeling processes observed in embryonic morphogenesis, wound repair, and cancer invasion. In this type of migration, cells move collectively in the form of sheets, strands, clusters, or ducts. Studies have found that preventing cancer cells from migrating and invading different tissues is a huge challenge in preventing and treating metastatic cancer. A scratch migration assay was used to evaluate the effects of MeJA-DMARE and MeJA-DMARE-phytosome on A549 cell migration. In addition, a wound closure assay was performed to evaluate the changes in A549 lung cancer cell migration (%) before and after treatment with MeJA-DMARE and MeJA-DMARE-phytosome (at a concentration of 250 μg / mL). The results are shown in Figure 3. Figure 10Figure 1 shows the migration of A549 lung cancer cells (A) and the amount of A549 lung cancer cell migration (B). The results show that MeJA-DMARE and MeJA-DMARE-phytosome treatment reduced the number of migrating cells, indicating that both inhibited the lateral movement of A549 cells. Notably, MeJA-DMARE-phytosome exhibited a stronger inhibitory effect than MeJA-DMARE.

[0089] Test Example 7

[0090] Regulatory effects of MeJA-DMARE and MeJA-DMARE-Phytosome on apoptosis gene expression in A549 lung cancer cells.

[0091] Figure 11 The figures show the regulatory effects of methyl jasmonate-induced extracts of the adventitious roots of Echinops chinensis and methyl jasmonate-induced phospholipid nanomaterials of the adventitious root extracts of Echinops chinensis on the expression of apoptotic genes in A549 lung cancer cells, wherein A and F are agarose gel electrophoresis bands of mRNA genes (from left to right: control, MeJA-DMARE, and MeJA-DMARE-phytosome), B is the mRNA expression level of the Caspase 3 gene, C is the mRNA expression level of the Bax gene, D is the mRNA expression level of the Bcl-2 gene, E is the mRNA expression level of the Caspase 9 gene, G is the mRNA expression level of the p38 gene, H is the mRNA expression level of the JNK gene, I is the mRNA expression level of the Nrf2 gene, J is the mRNA expression level of the HO-1 gene, and K is the mRNA expression level of the CAT gene.

[0092] Bcl-2 is an anti-apoptotic protein produced in response to DNA damage, which makes it play a key role in controlling cell death and serving as an indicator of intracellular genotoxic stress. Figure 11 As shown, MeJA-DMARE-phytosome significantly downregulated Bcl-2 expression by 0.24-fold. Conversely, the mRNA expression levels of Caspase 3, Caspase 9, and Bax were significantly upregulated by 2.37-fold, 4.53-fold, and 6.26-fold, respectively. The MeJA-DMARE-phytosome synthesized by the present invention can effectively inhibit the expression of apoptosis-related genes in lung cancer cells.

[0093] On the other hand, heme oxygenase-1 (HO-1) is well known for its strong induction response to various stressors and many cancer chemopreventive drugs. However, in human cancers, overexpression of HO-1 provides cancer cells with a growth advantage, making them more resistant to chemotherapy and photodynamic therapy. Nrf2 is a key transcription factor in the defense against oxidative stress, and activation of Nrf2 is an effective strategy to prevent cancer induced by exposure to environmental carcinogens. This activation is thought to be controlled by several upstream signaling kinases, including mitogen-activated protein kinases (MAPKs, such as p38 and JNK), which are able to regulate the activity of Nrf2. Figure 11 As shown in the figure, the mRNA expression levels of p38MAPK and JNK were the lowest in the control group. However, MeJA-DMARE-phytosome significantly upregulated these expressions in A549 lung cancer cells (p38MAPK and JNK increased by 4.52-fold and 3.40-fold, respectively). In addition, MeJA-DMARE-phytosome was able to significantly downregulate these expressions in A549 lung cancer cells (Nrf2, HO-1, and CAT decreased by 0.37-fold, 0.79-fold, and 0.44-fold, respectively).

[0094] It can be seen from the comprehensive test examples 5 to 7 that the MeJA-DMARE-Phytosome synthesized in the present invention has an anti-lung cancer effect.

[0095] Test Example 8

[0096] Stability testing

[0097] The in vitro stability of the MeJA-DMARE-phytosome prepared in the embodiment was tested in eight solvents: distilled water, 20 mM glycine-hydrochloric acid buffer (pH = 2.0), citrate-sodium citrate buffer (pH = 5.0), sodium phosphate buffer (pH = 7.0), tris-hydrochloric acid buffer (pH = 8.0), 5 wt% sodium chloride aqueous solution, 10 wt% sodium chloride aqueous solution, and 5% bovine serum albumin. The specific steps are as follows: 900 μL of solvent was mixed with 100 μL of a 10 mg / mL MeJA-DMARE-phytosome aqueous solution, and the mixture was incubated at 37° C. for one month. No obvious wavelength shift was observed by UV-visible spectroscopy, and the wavelength differences were all within 4 nm. This indicates that within one month, the physicochemical properties of the MeJA-DMARE-Phytosome prepared by the present invention were almost unchanged, indicating that the MeJA-DMARE-Phytosome has good stability.

[0098] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as within the scope of protection of the present invention.

Claims

1. A phospholipid nanomaterial derived from the adventitious root extract of the yellow lacquer tree induced by methyl jasmonate, characterized in that: The method uses an extract of the adventitious roots of the yellow lacquer tree induced by methyl jasmonate and a phospholipid bonded to the surface of the extract of the adventitious roots of the yellow lacquer tree induced by methyl jasmonate as raw materials; the bonding is through hydrogen bonding; The mass ratio of the extract of the adventitious roots of the yellow lacquer tree induced by methyl jasmonate to phospholipids is 1:0.5-3; The phospholipid is phosphatidylcholine; The preparation method of the adventitious root extract induced by methyl jasmonate comprises the following steps: mixing the adventitious root powder induced by methyl jasmonate with an extractant, performing extraction, concentrating the obtained extract and then drying to obtain the adventitious root extract induced by methyl jasmonate; the extractant is an ethanol aqueous solution, and the volume fraction of ethanol in the ethanol aqueous solution is 30-80%; The preparation method of the methyl jasmonate-induced phospholipid nanomaterial from the adventitious root extract of the yellow varnish wood comprises the following steps: mixing a solution of the adventitious root extract of the yellow varnish wood induced by methyl jasmonate and a phospholipid solution, and sequentially performing solvent removal, water dissolution, compounding, ultrasonic treatment, water dialysis, and drying to obtain the methyl jasmonate-induced phospholipid nanomaterial from the adventitious root extract of the yellow varnish wood; the compounding time is 1 to 2 hours, and the compounding is performed under shaking conditions at a shaking speed of 60 to 100 rpm.

2. The phospholipid nanomaterial of the adventitious root extract of Xanthoceras chinensis induced by methyl jasmonate according to claim 1, characterized in that: The solvent in the methyl jasmonate-induced yellow lacquerwood adventitious root extract solution is one or more of tetrahydrofuran, dichloromethane, chloroform, n-hexane, ethanol, ethyl acetate, cyclohexane, methanol, isopropanol, n-butanol and acetone; the concentration of the methyl jasmonate-induced yellow lacquerwood adventitious root extract solution is 1-10 g / L.

3. The phospholipid nanomaterial of the adventitious root extract of Xanthoceras chinensis induced by methyl jasmonate according to claim 1, characterized in that The solvent in the phospholipid solution is one or more of dichloromethane, chloroform and ethyl acetate; the concentration of the phospholipid solution is 1-10 g / L.

4. The phospholipid nanomaterial of the adventitious root extract of Xanthoceras chinensis induced by methyl jasmonate according to claim 1, characterized in that In the preparation of the phospholipid nanomaterials from the adventitious root extract of the jasmonate-induced methyl jasmonate, the mixing temperature is 25-45° C. and the mixing time is 1-30 minutes.

5. Use of the phospholipid nanomaterials of the adventitious root extract of Xanthoceras chinensis induced by methyl jasmonate according to any one of claims 1 to 4 in the preparation of anti-inflammatory drugs and / or anti-lung cancer drugs.

6. An anti-inflammatory and / or anti-lung cancer pharmaceutical composition comprising an active ingredient and a pharmaceutically acceptable excipient, wherein the active ingredient is the phospholipid nanomaterial derived from the adventitious root extract of the jasmonate-induced polyphylla according to any one of claims 1 to 4.

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