Chrysanthemum essential oil with function of improving dry age-related macular degeneration

Abstract

The invention belongs to the field of natural products, and provides chrysanthemum essential oil with a function of improving dry age-related macular degeneration. The chrysanthemum essential oil contains up to 168 compounds, more comprehensive biological activity support is provided for treatment, experiments prove that the essential oil can significantly optimize the retina structure of macular mice, specifically, the inner nuclear layer and the outer nuclear layer are closely arranged, the total thickness of the retina is significantly increased, the retinal pigment epithelium is more closely arranged, and the effect of improving the retina structure is achieved. Therefore, the polypeptide has the potential of improving the dry age-related macular degeneration function.

Description

Technical Field

[0001] The present invention belongs to the field of natural products, and particularly relates to a chrysanthemum essential oil having the function of improving dry age-related macular degeneration. Background Art

[0002] Existing technologies for the treatment of dry age-related macular degeneration (dry AMD) include drug therapy, photodynamic therapy, surgical treatment, and general treatment, but each technology has some drawbacks. Drug therapy may have limited effects and be accompanied by side effects; photodynamic therapy may cause damage to the normal retina and be costly; surgical treatment has high risks and is not effective for all patients; general treatment has slow effects and is difficult to directly improve the condition.

[0003] Although plant extracts have potential application value in the treatment of dry age-related macular degeneration (dry AMD), they face problems such as low bioavailability and delivery system limitations. The active ingredients in many plant extracts, such as carotenoids and polyphenols, etc., have low effective utilization rates after absorption and metabolism in the body. The key to treating dry AMD lies in effectively delivering these active ingredients to the retina. However, many plant extracts are difficult to penetrate the blood-retinal barrier. Summary of the Invention

[0004] To solve the above technical problems, the present invention proposes a chrysanthemum essential oil having the function of improving dry age-related macular degeneration. The active ingredients (such as terpene compounds, phenolic compounds) in this chrysanthemum essential oil are usually small molecule substances with small molecular weights, which are easier to penetrate the blood-retinal barrier. At the same time, many essential oil components are lipophilic and can interact with cell membranes, making them easier to be absorbed by cells, thereby increasing their concentration in retinal tissues. By the inhalation method, the essential oil components can quickly enter the blood circulation and improve bioavailability.

[0005] The technical solution of the present invention is as follows:

[0006] The present invention proposes a chrysanthemum essential oil having the function of improving dry age-related macular degeneration, and the chrysanthemum raw material is selected from any one or any two of chamomile, wild chrysanthemum, and blue tansy.

[0007] The preparation method of the above chrysanthemum essential oil mainly includes the following steps:

[0008] (1) Wash the chrysanthemum raw material, remove impurities, dry it at 35 - 50 °C, weigh 1 - 5 kg of the dried chrysanthemum raw material, and crush it to 40 meshes;

[0009] (2) Extraction: For the crushed chrysanthemum raw material, carry out CO extraction at 40 - 45 °C and 30 - 45 MPa 2Supercritical extraction is carried out for 1 - 2 h, and the extracted essential oil is filtered through a filter membrane to remove solid impurities, and then concentrated to remove residual moisture to obtain chrysanthemum essential oil.

[0010] The application of the above - mentioned chrysanthemum essential oil in the preparation of products for improving dry age - related macular degeneration is also the content that this invention focuses on protecting.

[0011] Preferably, during treatment, fractionated coconut oil is used to dilute the chrysanthemum essential oil to make a drug.

[0012] Preferably, the volume concentration of chrysanthemum essential oil in the drug is 1 - 10%.

[0013] Preferably, the volume concentration of chrysanthemum essential oil in the drug is 2 - 6%.

[0014] Preferably, the drug is used by olfactory inhalation.

[0015] Preferably, the specific method of olfactory inhalation: In a closed space, the chrysanthemum essential oil is placed at a distance of 0.1 - 1 m from the nostrils of the inhaler and maintained for 0.5 - 2 h.

[0016] Preferably, the frequency of olfactory inhalation is 1 time per day.

[0017] This invention has the following advantages and effects compared with the prior art:

[0018] (1) The chrysanthemum essential oil of this invention contains up to 168 compounds such as geraniol, α - caryophyllene, D(+)-camphor, trans - chrysanthenol, chrysanthenone, caryophyllin, palmitic acid, linoleic acid, γ - sitosterol, etc., covering various chemical components such as acids, phenyl esters, alcohols, terpenes, etc.; this complex chemical composition not only enriches the biological activity of the essential oil, but also endows it with unique curative effects, especially showing remarkable effects in improving macular degeneration; compared with the single or simply - composed essential oils in the prior art, the chrysanthemum essential oil of this invention can act on eye tissues more comprehensively and provide more effective treatment support;

[0019] (2) Experimental results show that after the macular mice are treated with chrysanthemum essential oil, the arrangement of the inner nuclear layer (INL) and outer nuclear layer (ONL) of the retina is closer, the total thickness of the retina increases significantly, showing significant improvement compared with the model group; particularly noteworthy is that the arrangement of the retinal pigment epithelium layer is also closer, and significant differences are shown at 7 days, 14 days, and 28 days after treatment; these experimental results not only verify the effectiveness of chrysanthemum essential oil in improving macular lesions, but also provide strong data support for its clinical application; compared with the prior art, the chrysanthemum essential oil of this invention shows more significant and lasting curative effects in treating macular lesions;

[0020] (3) From the perspective of practical applications, the chrysanthemum essential oil of the present invention has significant effects in treating macular lesions. The total retinal thickness of the model group was 106 μm, while that of the essential oil group reached 235.65 μm. This significant difference indicates that the chrysanthemum essential oil can significantly improve the retinal structure of macular mice. This means that for patients with macular lesions, using the chrysanthemum essential oil of the present invention for treatment is expected to achieve better vision recovery effects, thereby improving their quality of life. Compared with the prior art, the chrysanthemum essential oil of the present invention not only has higher efficacy but also has better safety and tolerance due to its natural ingredients, providing a more ideal treatment option for patients. Description of the Drawings

[0021] Figure 1 It is the GC-MS characterization diagram of the wild chrysanthemum essential oil sample in Example 1;

[0022] Figure 2 It is the HE staining diagram in Experimental Example 2;

[0023] Figure 3 It is the OCT diagram in Experimental Example 2. Detailed Embodiments

[0024] In order to enable those skilled in the art to better understand the present invention, the present invention will be further described below in conjunction with specific embodiments.

[0025] Example 1

[0026] The preparation method of wild chrysanthemum essential oil mainly includes the following steps:

[0027] (1) Wash the wild chrysanthemum with clean water, remove impurities, dry it at 40 °C, weigh 1000 grams of the dried wild chrysanthemum, and crush it to 40 mesh;

[0028] (2) Extraction: Load the crushed wild chrysanthemum into the extraction kettle to ensure uniform distribution, and perform supercritical extraction with CO at 42 °C and 40 MPa for 1.5 h. Filter the extracted essential oil through a filter membrane to remove solid impurities, and concentrate to remove residual moisture to obtain wild chrysanthemum essential oil. 2

[0029] Example 2

[0030] The preparation method of chamomile essential oil mainly includes the following steps:

[0031] (1) Wash the chamomile with clean water, remove impurities, dry it at 40 °C, weigh 1000 grams of the dried chamomile, and crush it to 40 mesh.

[0032] (2) Extraction: Load the crushed chamomile into the extraction kettle to ensure uniform distribution, and perform extraction with CO at 40 °C and 30 MPa2 Supercritical extraction was carried out for 1 h. The extracted essential oil was filtered through a filter membrane to remove solid impurities, and then concentrated to remove residual moisture to obtain chamomile essential oil.

[0033] Example 3

[0034] A method for preparing blue tansy essential oil mainly includes the following steps:

[0035] (1) Wash blue tansy with clean water to remove impurities, dry at 40 °C, weigh 1200 g of the dried blue tansy, and crush it to 40 mesh.

[0036] (2) Extraction: Load the crushed blue tansy into the extraction kettle to ensure uniform distribution, and carry out supercritical extraction with CO at 45 °C and 45 MPa 2 for 2 h. The extracted essential oil was filtered through a filter membrane to remove solid impurities, and then concentrated to remove residual moisture to obtain blue tansy essential oil.

[0037] Example 4

[0038] A method for preparing mixed chrysanthemum essential oil mainly includes the following steps:

[0039] (1) Wash two chrysanthemum raw materials, chamomile and wild chrysanthemum, with clean water to remove impurities, dry at 40 °C, weigh 500 g of the dried chamomile and 800 g of the dried wild chrysanthemum, and crush them to 40 mesh;

[0040] (2) Extraction: Load the crushed chrysanthemum raw materials into the extraction kettle to ensure uniform distribution, and carry out supercritical extraction with CO at 42 °C and 40 MPa 2 for 1.5 h. The extracted essential oil was filtered through a filter membrane to remove solid impurities, and then concentrated to remove residual moisture to obtain mixed chrysanthemum essential oil.

[0041] Example 5

[0042] A method for preparing mixed chrysanthemum essential oil mainly includes the following steps:

[0043] (1) Wash two chrysanthemum raw materials, chamomile and blue tansy, with clean water to remove impurities, dry at 40 °C, weigh 500 g of the dried chamomile and 800 g of the dried blue tansy, and crush them to 40 mesh;

[0044] (2) Extraction: Load the crushed chrysanthemum raw materials into the extraction kettle to ensure uniform distribution, and carry out supercritical extraction with CO at 42 °C and 40 MPa 2 for 1.5 h. The extracted essential oil was filtered through a filter membrane to remove solid impurities, and then concentrated to remove residual moisture to obtain mixed chrysanthemum essential oil.

[0045] Example 6

[0046] Preparation method of mixed chrysanthemum essential oil, mainly including the following steps:

[0047] (1) Wash two chrysanthemum raw materials, namely wild chrysanthemum and blue tansy, with clean water, remove impurities, dry at 40°C, weigh 500 g of dried wild chrysanthemum and 800 g of blue tansy, and crush them to 40 mesh;

[0048] (2) Extraction: Load the crushed chrysanthemum raw materials into the extraction kettle to ensure uniform distribution, and perform supercritical extraction with CO at 42°C and 40 MPa for 1.5 h. Filter the essential oil obtained by extraction through a filter membrane to remove solid impurities, and concentrate to remove residual moisture to obtain mixed chrysanthemum essential oil. 2

[0049] Experimental Example 1

[0050] Take the wild chrysanthemum essential oil sample in Example 1 and perform component analysis of the chrysanthemum essential oil by GC-MS:

[0051] The conditions for GC-MS analysis of chrysanthemum essential oil are as follows:

[0052] Shimadzu GC-MS-QP2020 NX - single quadrupole gas chromatography-mass spectrometry combined instrument, the chromatographic column is Agilent-DB-5ms, 30 m×0.25 mm×0.25 μm, the initial temperature of the column oven is 50°C, the initial time is 4 min, the heating rate is 5°C / min, the termination temperature is 260°C, the termination time is 10 min, the total flow rate is 1 mL / min, and it is non-split. The ion source temperature is 230°C, the interface temperature is 280°C, and the mass number detection range is 29 - 500. The GC-MS analysis and test characterization results are shown in the appendix Figure 1 , and the chemical composition of the chrysanthemum essential oil sample is shown in Table 1 in detail:

[0053] Table 1 Chemical composition of chrysanthemum essential oil sample

[0054]

[0055]

[0056]

[0057]

[0058]

[0059] ​From the chemical composition of the above chrysanthemum essential oil sample, it can be seen that the chrysanthemum essential oil of the present invention is composed of 168 compounds such as geraniol, α-caryophyllene alcohol, D(+)-camphor, trans-chrysanthenol, chrysanthenone, caryophyllin, palmitic acid, linoleic acid, γ-sitosterol, etc. It contains acids, phenyl esters, alcohols, terpenes, etc. The chemical composition is complex and has the effect of improving macular degeneration.

[0060] Experimental Example 2

[0061] 1. Animal experiment design

[0062] Male ICR mice (five weeks old) from Vital River were used. NaIO 3 solution was injected into the tail vein to establish a dry macular degeneration model, and the remission effect on macular mice was detected after different treatment interventions. ICR male white mice at five weeks old with a body weight of 24 - 26 g were selected.

[0063] The ICR mice were raised in the same breeding environment with a room temperature of (20 ± 2) °C, a relative humidity of 50% - 65%, and good ventilation. They were fed with standard mouse feed, allowed to freely eat and drink water, and adaptively fed for one week.

[0064] The selected mice were randomly grouped according to body weight, and a blank group, a model group, and a chrysanthemum essential oil group were set up, with 6 mice in each group.

[0065] First, the mice in the model group and the chrysanthemum essential oil group were used to establish a dry eye model by injecting NaIO 3 solution into the tail vein. 40 mg / kg NaIO 3 solution was injected into the tail vein of the mice.

[0066] Each group was then treated as follows, and the remission effect on the macula was evaluated on the 7th, 14th, and 28th days after treatment.

[0067] (1) Blank group: Treated with fractionated coconut oil;

[0068] (2) Model group: Treated with fractionated coconut oil;

[0069] (3) Essential oil group: The chrysanthemum essential oil obtained in Example 1 was diluted to a volume concentration of 3% with fractionated coconut oil, and in a closed space, the mice were treated by olfactory inhalation for 0.5 h, with an action distance of 0.1 m and an action frequency of 1 time / day.

[0070] 2. Histopathological analysis (evaluation of remission effect on the macula)

[0071] The mouse eyeballs were taken at 7 days, 14 days, and 28 days after treatment respectively, fixed with 4% paraformaldehyde, embedded in paraffin, and sectioned into 5 μm thickness. The paraffin sections were stained with hematoxylin & eosin staining kit (Auragene, Hunan, China) according to the instruction manual. All stained photos were taken through an optical microscope (Nikon, Melville, NY) to observe the status of the retinal area, see Figure 2 .

[0072] From Figure 2 the results, it can be seen that after the macular mice were treated with chrysanthemum essential oil, the inner nuclear layer (INL) and outer nuclear layer (ONL) of the retina were arranged more closely compared with the model group, and the total retinal thickness was larger than that of the model group. The retinal pigment epithelium layer was arranged more closely compared with the model group, and there were significant differences at 7 days, 14 days, and 28 days after treatment. It shows that the treatment method of chrysanthemum essential oil has the effect of improving the lesion.

[0073] 3. Optical coherence tomography analysis (evaluation of the effect of alleviating macular)

[0074] After the administration cycle of the mice ended, they were anesthetized by inhaling isoflurane. Compound tropicamide eye drops were dropped into the eyes of the mice to dilate the pupils for 10 minutes, and then the mice were fixed. Medical sodium hyaluronate was used to protect the cornea. After wearing the corneal contact lens, OCT detection was carried out to observe the fundus of the mice. The cross-sectional image of the mouse retina was obtained with the optic disc as the marker, and the retinal thickness at 1000 μm away from the optic nerve was seen Figure 3 .

[0075] According to Figure 3 the results, measurement and analysis were carried out with IMAGE J software, as shown in Table 2.

[0076] Table 2 Quantitative analysis of the thickness of each cell layer of the retina

[0077]

[0078]

[0079] The total retinal thickness of the model group was 106 μm; the total retinal thickness of the essential oil group was 235.65 μm. It can be seen that after the macular mice were treated with chrysanthemum essential oil, the total retinal thickness was larger than that of the model group. It shows that the treatment method of the chrysanthemum essential oil group has the effect of improving macular degeneration.

[0080] In summary, the chrysanthemum essential oil provided by the present invention can effectively improve the function of dry age-related macular degeneration.

[0081] The above are only the preferred embodiments of the present invention, and do not limit the patent scope of the present invention accordingly. All equivalent changes and modifications made according to the scope of the present invention should still fall within the scope covered by the present invention.

Claims

1. A chrysanthemum essential oil having the function of improving dry age-related macular degeneration, characterized in that: The chrysanthemum raw material of the chrysanthemum essential oil is selected from any one or any two of chamomile, wild chrysanthemum and blue tansy.

2. The method for preparing chrysanthemum essential oil according to claim 1, characterized in that: The main steps include: (1) Wash the chrysanthemum raw material, remove impurities, dry it at 35-50°C, weigh 1-5 kg ​​of the dried chrysanthemum raw material, and crush it into 40 mesh; (2) Extraction: The crushed chrysanthemum raw material is subjected to CO2 supercritical extraction at 40-45°C and 30-45MPa for 1-2h. The essential oil obtained by the extraction is filtered through a filter membrane to remove solid impurities and concentrated to remove residual water to obtain chrysanthemum essential oil.

3. Use of the chrysanthemum essential oil according to any one of claims 1 to 2 in preparing products for improving dry age-related macular degeneration.

4. The use as claimed in claim 3, characterized in that Chrysanthemum essential oil is diluted with fractionated coconut oil to make a medicine.

5. The use as claimed in claim 4, characterized in that The volume concentration of chrysanthemum essential oil in the medicine is 1-10%.

6. The use as claimed in claim 4, characterized in that The volume concentration of chrysanthemum essential oil in the medicine is 2-6%.

7. The use as claimed in claim 4, characterized in that The drug is administered by inhalation.

8. The use as claimed in claim 7, characterized in that The specific method of using the sniffing inhalation is as follows: in a closed space, the chrysanthemum essential oil is placed at a distance of 0.1 to 1 m from the nostrils of the ingestor and maintained for 0.5 to 2 hours.

9. The use as claimed in claim 7, characterized in that The sniffing intake frequency is once per day.

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