A traditional Chinese medicine fermentation composition for relieving visual fatigue and preparation method thereof

Abstract

The present invention belongs to the technical field of traditional Chinese medicine, and specifically relates to a traditional Chinese medicine fermentation composition for relieving visual fatigue and a preparation method thereof. The present invention discloses a preparation method of the traditional Chinese medicine fermentation composition for relieving visual fatigue, comprising: using a composite enzyme to hydrolyze the traditional Chinese medicine components, and then fermenting the hydrolyzed traditional Chinese medicine components with a mixed bacteria composed of Lactobacillus rhamnosus, Saccharomyces cerevisiae and Aspergillus niger. The prepared traditional Chinese medicine fermentation composition has been verified to be safe, non-irritating and has better efficacy; and the massage cream prepared using the traditional Chinese medicine fermentation composition has a significant effect of relieving visual fatigue. The preparation method of the traditional Chinese medicine fermentation composition described in this application is simple, can solve the problems of cumbersome and complicated, long treatment course and slow effect in the existing traditional Chinese medicine treatment of visual fatigue, and has good clinical application value.

Description

Technical Field

[0001] The invention belongs to the technical field of traditional Chinese medicines, and particularly relates to a traditional Chinese medicine fermentation composition for relieving visual fatigue and a preparation method thereof. Background Art

[0002] Visual fatigue, also known as eye strain, refers to eye fatigue caused by excessive use of the eyes when engaged in close-up work or study. The main clinical manifestations are blurred vision, double images, intolerance to prolonged viewing, dry and uncomfortable eyes, soreness and tearing, headache and dizziness. It is a syndrome based on the patient's subjective eye symptoms and is an intertwined combination of ocular or systemic organic factors and mental factors. Therefore, it is also called eye fatigue syndrome and belongs to the category of psychosomatic medicine.

[0003] Western medicine primarily treats visual fatigue with symptomatic and corrective treatments, typically using sedatives, neurotrophic agents, and topical beta-blockers. Additionally, surgery, prism correction, or visual training may be used to treat visual fatigue. While these methods are relatively effective, they generally have significant side effects and are relatively limited in scope, failing to meet the needs of preventing and treating visual fatigue syndrome. Traditional Chinese medicine offers a variety of methods for treating visual fatigue, including acupuncture, acupoint massage, Chinese herbal compresses, and Chinese herbal atomization, all of which can achieve good results. Furthermore, because they fall within the scope of TCM external treatments, are convenient and flexible to use, and offer significant efficacy, they are favored by a large number of patients with visual fatigue, demonstrating the advantages of TCM in the diagnosis and treatment of visual fatigue.

[0004] In recent years, with the advent of the "screen-reading era," people have been spending increasingly extended periods using various electronic devices, such as mobile phones, tablets, and computers. This puts a high workload on the eyes, exacerbating overuse and significantly increasing the likelihood of eye diseases. However, current traditional Chinese medicine treatments for visual fatigue often suffer from drawbacks such as cumbersome and complex procedures, long treatment cycles, and slow effectiveness. Therefore, providing a convenient, simple-to-use, and rapidly effective traditional Chinese medicine product is a pressing need. Summary of the Invention

[0005] In response to the above defects or improvement needs of the prior art, the present invention provides a traditional Chinese medicine fermentation composition for relieving visual fatigue and a preparation method thereof, as well as a massage cream prepared using the traditional Chinese medicine fermentation composition, to solve the problems of cumbersome and complicated, long treatment course and slow effect in the existing traditional Chinese medicine treatment of visual fatigue.

[0006] The first aspect of the present invention aims to provide a method for preparing a traditional Chinese medicine fermentation composition for alleviating visual fatigue.

[0007] The purpose of the present invention can be achieved through the following technical solutions:

[0008] The first aspect of the present invention provides a method for preparing a traditional Chinese medicine fermentation composition for relieving visual fatigue, comprising:

[0009] S1. Weigh each raw material in the Chinese medicine composition according to parts by weight, grind, and sieve to obtain a mixed powder; add sterile deionized water to the mixed powder, stir evenly, and obtain a mixed solution I;

[0010] S2, adding pectinase and cellulase to the mixed solution I, and performing ultrasound-assisted enzymolysis; after the enzymolysis is completed, heating the solution to inactivate the enzymes to obtain a mixed solution II;

[0011] S3, respectively activating Lactobacillus rhamnosus, Saccharomyces cerevisiae, and Aspergillus niger with liquid culture medium, and inoculating the activated Lactobacillus rhamnosus, Saccharomyces cerevisiae, and Aspergillus niger into mixed solution II for mixed fermentation;

[0012] S4. After the fermentation is completed, the filtrate is filtered and concentrated under reduced pressure to obtain a concentrated solution; the concentrated solution is sterilized at high temperature to obtain the traditional Chinese medicine fermentation composition.

[0013] Preferably, the Lactobacillus rhamnosus is deposited under the number CGMCC 1.3724; the Saccharomyces cerevisiae is deposited under the number CGMCC 2.1527; and the Aspergillus niger is deposited under the number CGMCC 3.316.

[0014] Preferably, the traditional Chinese medicine composition comprises the following raw materials in parts by weight: 3-15 parts of wolfberry, 1-10 parts of chrysanthemum, 6-20 parts of Xing'an Cimicifuga, 1-10 parts of Ligustrum lucidum, 5-16 parts of Ligustrum lucidum, 5-16 parts of safflower, 1-10 parts of coptis chinensis, 1-10 parts of equisetum, 1-10 parts of gardenia, 1-10 parts of tribulus terrestris, 5-16 parts of dodder seed, 6-18 parts of cassia seed, 1-10 parts of buddlejac, 1-10 parts of white peony root, and 1-10 parts of mulberry leaf.

[0015] Further preferably, the Chinese medicine composition comprises the following raw materials in parts by weight: 10 parts of wolfberry, 4 parts of chrysanthemum, 13 parts of Xing'an Cimicifuga, 5 parts of Ligustrum lucidum, 10 parts of Ligustrum lucidum, 10 parts of safflower, 4 parts of Coptis chinensis, 5 parts of Equisetum arvense, 5 parts of Gardenia jasminoides, 5 parts of Tribulus terrestris, 11 parts of Cuscuta chinensis, 12 parts of Cassia tora, 4 parts of Buddleja bud, 4 parts of white peony root, and 4 parts of mulberry leaf.

[0016] Preferably, in step S1, the mesh number of the sieve is 200-300 mesh, and the material-liquid ratio of the mixed powder of the traditional Chinese medicine composition to sterile deionized water is 1:5-7.

[0017] Preferably, in step S2, the mass ratio of pectinase to cellulase is 2:1, the total enzyme amount is 3-5wt% of the mass of the mixed solution I, the enzymatic hydrolysis temperature is 45-55°C, the pH is 4.5-5.5, the ultrasonic frequency is 25-35kHz, and the time is 40-50min; the enzyme inactivation treatment temperature is 90-100°C, and the enzyme inactivation treatment time is 20-30min.

[0018] Preferably, in step S3, the volume ratio of Lactobacillus rhamnosus, Saccharomyces cerevisiae and Aspergillus niger is 1-2:1:1-1.5.

[0019] Further preferably, in step S3, the volume ratio of Lactobacillus rhamnosus, Saccharomyces cerevisiae and Aspergillus niger is 1.5:1:1.2.

[0020] Preferably, the total amount of bacteria added is 5-7% of the volume of the mixed solution II.

[0021] Preferably, in step S3, the fermentation temperature is 30-34° C., the fermentation pH is 5.0-6.0, the sterile air ventilation volume is 5 L / min, the rotation speed is 200-250 rpm, and the fermentation time is 48-72 h.

[0022] Preferably, in step S4, the filtrate is concentrated under reduced pressure to 80% of the mass of the original filtrate, the temperature for the concentration is 40-50° C., the pressure for the concentration is -0.06 to -0.08 MPa, the sterilization temperature is 121° C., and the sterilization time is 20-30 min.

[0023] The purpose of the second aspect of the present invention is to provide a Chinese medicinal fermentation composition prepared by the preparation method of the first aspect of the present invention.

[0024] The third aspect of the present invention aims to provide a massage cream for relieving visual fatigue, comprising the Chinese medicinal fermentation composition described in the second aspect.

[0025] Preferably, the massage cream comprises the following raw materials in percentage by weight: 5.0-20.0% of Chinese medicinal fermentation composition, 6.5-8.5% of emulsifier, 3.0-8.0% of excipient, 8.0-13.0% of emollient, 0.18-0.68% of conditioner, 0.3-2.3% of suspending agent, 0.1-1.0% of covering agent, 10.5-14.5% of moisturizer, and 0.05-0.15% of chelating agent.

[0026] Beneficial effects of the present invention:

[0027] The present invention hydrolyzes Chinese medicinal ingredients by a composite enzyme to release the active substances in Chinese medicinal ingredients, and then ferments the Chinese medicinal ingredients after enzymatic hydrolysis by a mixed bacteria composed of Lactobacillus rhamnosus, Saccharomyces cerevisiae and Aspergillus niger. Aspergillus niger can produce cellulose to help Chinese medicinal ingredients further release active substances; Lactobacillus rhamnosus is a safe probiotic, and the proteins or metabolites it secretes have multiple potential health benefits such as enhancing immune function and inhibiting bacteria; yeast can cooperate with lactobacillus to reproduce and complement each other in nutrition, promoting the proliferation of lactobacillus. In order to improve the therapeutic effect of a specific Chinese medicinal composition, the inventors screened and verified a large number of experiments in the early stage, and finally obtained a composite bacteria composed of Lactobacillus rhamnosus, Saccharomyces cerevisiae and Aspergillus niger. The above bacteria have good synergy during the fermentation process, making the prepared Chinese medicinal fermentation composition safe and non-irritating and better in efficacy after experimental verification, and the massage cream prepared using the Chinese medicinal fermentation composition has a significant effect of relieving visual fatigue. The preparation method of the Chinese medicinal fermentation composition described in this application is simple, can solve the problems of cumbersome and complex, long treatment course and slow effect when existing Chinese medicine is used to treat visual fatigue, and has good clinical application value. DETAILED DESCRIPTION

[0028] In order to further illustrate the technical means and effects adopted by the present invention to achieve the predetermined purpose of the invention, the specific implementation methods, structures, features and effects of the present invention are described in detail below in combination with the embodiments.

[0029] In the following examples, experimental methods without specific conditions were generally performed under conventional conditions or those recommended by the manufacturer. Materials and reagents used were commercially available unless otherwise specified.

[0030] Cellulase was purchased from Shanghai Yuanye Biotechnology Co., Ltd. with an enzyme activity of 50 u / mg;

[0031] Pectinase was purchased from Shanghai Yuanye Biotechnology Co., Ltd. with an enzyme activity of 50,000 u / g;

[0032] The viable counts of Lactobacillus rhamnosus, Saccharomyces cerevisiae and Aspergillus niger involved in the present invention are all 1×10 7 CFU / mL.

[0033] Example 1

[0034] A method for preparing a traditional Chinese medicine fermentation composition for relieving visual fatigue, the method comprising the following steps:

[0035] S1. Weigh each raw material in the Chinese medicine composition according to parts by weight, grind, and sieve to obtain a mixed powder; add sterile deionized water to the mixed powder, stir evenly, and obtain a mixed solution I;

[0036] S2, adding pectinase and cellulase to the mixed solution I, and performing ultrasound-assisted enzymolysis; after the enzymolysis is completed, heating the solution to inactivate the enzymes to obtain a mixed solution II;

[0037] S3, respectively activating Lactobacillus rhamnosus, Saccharomyces cerevisiae, and Aspergillus niger with liquid culture medium, and inoculating the activated Lactobacillus rhamnosus, Saccharomyces cerevisiae, and Aspergillus niger into mixed solution II for mixed fermentation;

[0038] S4, after the fermentation is completed, the filtrate is filtered and concentrated under reduced pressure to obtain a concentrate; the concentrate is sterilized at high temperature to obtain the Chinese medicine fermentation composition;

[0039] Among them, the traditional Chinese medicine composition includes the following raw materials in parts by weight: 3 parts of wolfberry, 1 part of chrysanthemum, 6 parts of Xing'an Cimicifuga, 1 part of Ligustrum lucidum, 5 parts of Ligustrum lucidum, 5 parts of safflower, 1 part of Coptis chinensis, 1 part of Equisetum arvense, 1 part of Gardenia jasminoides, 1 part of Tribulus terrestris, 5 parts of Cuscuta chinensis, 6 parts of Cassia tora, 1 part of Buddleja bud, 1 part of white peony root, and 1 part of mulberry leaf.

[0040] In the step S1, the mesh number of the sieve is 200 mesh, and the material-liquid ratio of the mixed powder of the traditional Chinese medicine composition to sterile deionized water is 1:5.

[0041] In step S2, the mass ratio of pectinase to cellulase is 2:1, the total enzyme amount is 3wt% of the mass of the mixed solution I, the enzymatic hydrolysis temperature is 45°C, the pH is 4.5, the ultrasonic frequency is 25kHz, and the time is 40min; the enzyme inactivation temperature is 90°C, and the enzyme inactivation time is 20min.

[0042] In step S3, the volume ratio of Lactobacillus rhamnosus, Saccharomyces cerevisiae and Aspergillus niger is 1:1:1.5; and the total amount of bacteria added is 5% of the volume of the mixed solution II.

[0043] The Lactobacillus rhamnosus has a preservation number of CGMCC 1.3724; the Saccharomyces cerevisiae has a preservation number of CGMCC 2.1527; and the Aspergillus niger has a preservation number of CGMCC 3.316.

[0044] In step S3, the fermentation temperature is 30° C., the fermentation pH is 5.0, the sterile air ventilation volume is 5 L / min, the rotation speed is 200 rpm, and the fermentation time is 48 h.

[0045] In the step S4, the filtrate is concentrated under reduced pressure to 80% of the mass of the original filtrate, the temperature of the concentrated under reduced pressure is 40° C., the pressure of the concentrated under reduced pressure is -0.06 MPa; the sterilization temperature is 121° C., and the sterilization time is 20 min.

[0046] Example 2

[0047] A method for preparing a traditional Chinese medicine fermentation composition for relieving visual fatigue, the method comprising the following steps:

[0048] S1. Weigh each raw material in the Chinese medicine composition according to parts by weight, grind, and sieve to obtain a mixed powder; add sterile deionized water to the mixed powder, stir evenly, and obtain a mixed solution I;

[0049] S2, adding pectinase and cellulase to the mixed solution I, and performing ultrasound-assisted enzymolysis; after the enzymolysis is completed, heating the solution to inactivate the enzymes to obtain a mixed solution II;

[0050] S3, respectively activating Lactobacillus rhamnosus, Saccharomyces cerevisiae, and Aspergillus niger with liquid culture medium, and inoculating the activated Lactobacillus rhamnosus, Saccharomyces cerevisiae, and Aspergillus niger into mixed solution II for mixed fermentation;

[0051] S4. After the fermentation is completed, the filtrate is filtered and concentrated under reduced pressure to obtain a concentrated solution; the concentrated solution is sterilized at high temperature to obtain the traditional Chinese medicine fermentation composition.

[0052] Among them, the traditional Chinese medicine composition includes the following raw materials in parts by weight: 10 parts of wolfberry, 4 parts of chrysanthemum, 13 parts of Xing'an Cimicifuga, 5 parts of Ligustrum lucidum, 10 parts of Ligustrum lucidum, 10 parts of safflower, 4 parts of Coptis chinensis, 5 parts of Equisetum arvense, 5 parts of Gardenia jasminoides, 5 parts of Tribulus terrestris, 11 parts of Cuscuta chinensis, 12 parts of Cassia tora, 4 parts of Buddleja bud, 4 parts of white peony root, and 4 parts of mulberry leaf.

[0053] In the step S1, the mesh number of the sieve is 250 mesh, and the material-liquid ratio of the mixed powder of the traditional Chinese medicine composition to the sterile deionized water is 1:6.

[0054] In step S2, the mass ratio of pectinase to cellulase is 2:1, the total enzyme amount is 4 wt% of the mass of the mixed solution I, the enzymatic hydrolysis temperature is 50°C, the pH is 5.0, the ultrasonic frequency is 30 kHz, and the time is 45 min; the enzyme inactivation temperature is 95°C, and the enzyme inactivation time is 25 min.

[0055] In step S3, the volume ratio of Lactobacillus rhamnosus, Saccharomyces cerevisiae and Aspergillus niger is 1.5:1:1.2; and the total amount of bacteria added is 6% of the volume of the mixed solution II.

[0056] The Lactobacillus rhamnosus has a preservation number of CGMCC 1.3724; the Saccharomyces cerevisiae has a preservation number of CGMCC 2.1527; and the Aspergillus niger has a preservation number of CGMCC 3.316.

[0057] In step S3, the fermentation temperature is 32° C., the fermentation pH is 5.5, the sterile air ventilation volume is 5 L / min, the rotation speed is 220 rpm, and the fermentation time is 60 h.

[0058] In step S4, the filtrate is concentrated under reduced pressure to 80% of the mass of the original filtrate, the temperature for the concentration is 45° C., the pressure for the concentration is -0.07 MPa, and the sterilization temperature is 121° C., and the sterilization time is 25 min.

[0059] Example 3

[0060] A method for preparing a traditional Chinese medicine fermentation composition for relieving visual fatigue, the method comprising the following steps:

[0061] S1. Weigh each raw material in the Chinese medicine composition according to parts by weight, grind, and sieve to obtain a mixed powder; add sterile deionized water to the mixed powder, stir evenly, and obtain a mixed solution I;

[0062] S2, adding pectinase and cellulase to the mixed solution I, and performing ultrasound-assisted enzymolysis; after the enzymolysis is completed, heating the solution to inactivate the enzymes to obtain a mixed solution II;

[0063] S3, respectively activating Lactobacillus rhamnosus, Saccharomyces cerevisiae, and Aspergillus niger with liquid culture medium, and inoculating the activated Lactobacillus rhamnosus, Saccharomyces cerevisiae, and Aspergillus niger into mixed solution II for mixed fermentation;

[0064] S4, after the fermentation is completed, the filtrate is filtered and concentrated under reduced pressure to obtain a concentrate; the concentrate is sterilized at high temperature to obtain the Chinese medicine fermentation composition;

[0065] Among them, the traditional Chinese medicine composition includes the following raw materials in parts by weight: 15 parts of wolfberry, 10 parts of chrysanthemum, 20 parts of Xing'an Cimicifuga, 10 parts of Ligustrum lucidum, 16 parts of Ligustrum lucidum, 16 parts of safflower, 10 parts of coptis chinensis, 10 parts of horsetail, 10 parts of gardenia, 10 parts of tribulus terrestris, 16 parts of dodder seed, 18 parts of cassia seed, 10 parts of buddlejac, 10 parts of white peony root, and 10 parts of mulberry leaf.

[0066] In the step S1, the mesh number of the sieve is 300 mesh, and the material-liquid ratio of the mixed powder of the traditional Chinese medicine composition to sterile deionized water is 1:7.

[0067] In step S2, the mass ratio of pectinase to cellulase is 2:1, the total enzyme amount is 5wt% of the mass of the mixed solution I, the enzymatic hydrolysis temperature is 55°C, the pH is 5.5, the ultrasonic frequency is 35kHz, and the time is 50min; the enzyme inactivation temperature is 100°C, and the enzyme inactivation time is 30min.

[0068] In step S3, the volume ratio of Lactobacillus rhamnosus, Saccharomyces cerevisiae and Aspergillus niger is 2:1:1; and the total amount of bacteria added is 7% of the volume of the mixed solution II.

[0069] The Lactobacillus rhamnosus has a preservation number of CGMCC 1.3724; the Saccharomyces cerevisiae has a preservation number of CGMCC 2.1527; and the Aspergillus niger has a preservation number of CGMCC 3.316.

[0070] In step S3, the fermentation temperature is 34° C., the fermentation pH is 6.0, the sterile air ventilation volume is 5 L / min, the rotation speed is 250 rpm, and the fermentation time is 72 h.

[0071] In step S4, the filtrate is concentrated under reduced pressure to 80% of the mass of the original filtrate, the temperature for the concentration is 50° C., the pressure for the concentration is -0.08 MPa, and the sterilization temperature is 121° C., and the sterilization time is 30 min.

[0072] Comparative Example 1

[0073] Compared with Example 2, the only difference is that in Comparative Example 1, Lactobacillus rhamnosus is not added, and its reduced volume is proportionally distributed to the volumes of Saccharomyces cerevisiae and Aspergillus niger. Other conditions are the same as in Example 2.

[0074] Comparative Example 2

[0075] Compared with Example 2, the only difference is that in Comparative Example 2, no cerevisiae yeast is added, and the reduced volume is proportionally distributed to the volumes of Lactobacillus rhamnosus and Aspergillus niger. Other conditions are the same as in Example 2.

[0076] Comparative Example 3

[0077] Compared with Example 2, the only difference is that the volume ratio of Lactobacillus rhamnosus, Saccharomyces cerevisiae and Aspergillus niger is 0.5:1:2, and other conditions are the same as those in Example 2.

[0078] Comparative Example 4

[0079] Compared with Example 2, the only difference is that the volume ratio of Lactobacillus rhamnosus, Saccharomyces cerevisiae and Aspergillus niger is 2.5:1:0.5, and other conditions are the same as those in Example 2.

[0080] Comparative Example 5

[0081] Compared with Example 2, the only difference is that Lactobacillus rhamnosus with the deposit number ATCC 7469 is used instead of Lactobacillus rhamnosus with the deposit number CGMCC 1.3724, and other conditions are the same as those in Example 2.

[0082] Comparative Example 6

[0083] Based on Example 2, the only difference is that the Saccharomyces cerevisiae with the deposit number CGMCC 2.3854 is used instead of the Saccharomyces cerevisiae with the deposit number CGMCC 2.1527, and other conditions are the same as those in Example 2.

[0084] Comparative Example 7

[0085] Based on Example 2, the only difference is that Aspergillus niger with the deposit number CGMCC 3.4309 is used instead of Aspergillus niger with the deposit number CGMCC 3.316, and other conditions are the same as those in Example 2.

[0086] Comparative Example 8

[0087] A method for preparing a traditional Chinese medicine composition for relieving visual fatigue, comprising the following steps:

[0088] S1. Weigh each raw material in the Chinese medicine composition according to parts by weight, grind, and sieve to obtain a mixed powder; add sterile deionized water to the mixed powder, stir evenly, and obtain a mixed solution I;

[0089] S2, adding pectinase and cellulase to the mixed solution I, and performing ultrasound-assisted enzymolysis; after the enzymolysis is completed, heating the solution to inactivate the enzymes to obtain a mixed solution II;

[0090] S3, filtering the mixed solution II, and concentrating the filtrate under reduced pressure to obtain a concentrated solution, thereby obtaining the Chinese medicine composition.

[0091] Among them, the traditional Chinese medicine composition includes the following raw materials in parts by weight: 10 parts of wolfberry, 4 parts of chrysanthemum, 13 parts of Xing'an Cimicifuga, 5 parts of Ligustrum lucidum, 10 parts of Ligustrum lucidum, 10 parts of safflower, 4 parts of Coptis chinensis, 5 parts of Equisetum arvense, 5 parts of Gardenia jasminoides, 5 parts of Tribulus terrestris, 11 parts of Cuscuta chinensis, 12 parts of Cassia tora, 4 parts of Buddleja bud, 4 parts of white peony root, and 4 parts of mulberry leaf.

[0092] In the step S1, the mesh number of the sieve is 250 mesh, and the material-liquid ratio of the mixed powder of the traditional Chinese medicine composition to the sterile deionized water is 1:6.

[0093] In step S2, the mass ratio of pectinase to cellulase is 2:1, the total enzyme amount is 4 wt% of the mass of the mixed solution I, the enzymatic hydrolysis temperature is 50°C, the pH is 5.0, the ultrasonic frequency is 30 kHz, and the time is 45 min; the enzyme inactivation temperature is 95°C, and the enzyme inactivation time is 25 min.

[0094] In step S3, the filtrate is concentrated under reduced pressure to 80% of the mass of the original filtrate, the temperature of the concentrated under reduced pressure is 45° C., and the pressure of the concentrated under reduced pressure is -0.07 MPa.

[0095] Effect Example 1 Acute eye irritation test

[0096] With reference to the Technical Specifications for Safety of Cosmetics and the acute eye irritation test of patent CN116492383B, the irritation of the compositions prepared in Examples 1-3 and Comparative Examples 1-8 was verified.

[0097] Experimental animals: Female, adult New Zealand rabbits with healthy eyes, weighing 2.0-2.5 kg, were fed with a standard rabbit feed and housed in single cages in a pathogen-free environment at a constant temperature of 20-26°C and a relative humidity of 40%-70%.

[0098] The specific method is as follows: healthy adult New Zealand rabbits were used, and the rabbits were acclimated to the experimental animal room environment for 7 days before the test. 33 rabbits without eye irritation symptoms, corneal defects, and conjunctival damage were selected and randomly divided into 11 groups, with 3 rabbits in each group, to test the compositions prepared in Examples 1-3 and Comparative Examples 1-8, respectively. The lower eyelid of one rabbit's eye was gently pulled open, and 0.1 mL of the test substance (the composition prepared in accordance with Examples 1-3 and Comparative Examples 1-8, respectively) was instilled into the conjunctival sac. The upper and lower eyelids were passively closed for 1 second to prevent loss of the test substance. The other eye was left untreated as a self-control. The eye was not rinsed within 24 hours after the test substance was instilled. If necessary, it could be rinsed at 24 hours. If the above test results showed that the test substance was irritating, 3 rabbits were selected to conduct a flushing effect test. That is, 30 seconds after the test substance was instilled into the rabbit eye, the eye was rinsed for at least 30 seconds with sufficient, fast-flowing water that did not cause eye damage. The eyes of the animals were examined 1, 24, 48, 72 hours after the instillation of the composition, as well as on the 4th and 7th days. If no irritation reaction occurred after 72 hours, the test was terminated.

[0099] The evaluation was performed according to Table 1. The sum of the irritation response scores for the cornea, iris, conjunctiva, and secretions of each rabbit was used to obtain the total score for the irritation response of the rabbit eye. The sum of the total score for the irritation response of each rabbit eye was divided by the number of rabbits to obtain the final score for the irritation of the composition to the rabbits.

[0100] Conclusion: The test results are shown in Table 2. When the eyes of the 11 groups of rabbits were examined at 1 hour, 24 hours, 48 ​​hours and 72 hours, no corneal, iris or conjunctival lesions or congestion were observed, and there was no secretion. The total eye irritation score was zero, indicating that the compositions prepared in Examples 1-3 and Comparative Examples 1-8 were safe and non-irritating.

[0101] Table 1 Scoring criteria for acute eye irritation

[0102]

[0103]

[0104] Table 2 Integral results of acute eye irritation test

[0105]

[0106]

[0107] Effect Example 2 Efficacy Verification

[0108] The efficacy of the compositions prepared in Examples 1-3 and Comparative Examples 1-8 was verified, and the specific steps were as follows:

[0109] Experimental animals: 112 healthy male New Zealand rabbits, weighing 2.0-2.5 kg, were fed with ordinary feed and housed in single cages in a pathogen-free environment at a constant temperature of 20-26°C and a relative humidity of 40%-70%.

[0110] Grouping: 112 New Zealand rabbits were randomly divided into a blank group, a model group, a positive control group, Example 1-3 groups, and Comparative Example 1-8 groups, for a total of 14 groups, with 8 rabbits in each group, for a total of 112 rabbits.

[0111] Visual fatigue animal model: Except for the blank group, the other groups were placed under a day-night light cycle, with 35 lux (illuminance) during the day and 0 lux at night for 6 consecutive days.

[0112] Grouped administration: After modeling, rabbits in Examples 1-3 and Comparative Examples 1-8 were orally administered with the composition prepared in the corresponding groups at a dose of 5 mL / rabbit. Rabbits in the positive control group were orally administered with Qiju Dihuang oral liquid. Rabbits in the blank group and the model group were orally administered with an equal amount of normal saline. The administration was performed once a day for 4 consecutive weeks.

[0113] Index detection: 24 hours after the last administration, the experimental animals were sacrificed, the bilateral eyeballs were removed, and the ciliary bodies were carefully separated under an operating microscope. Rinse with physiological saline, dry with filter paper, and weigh. Prepare with an internal tissue homogenizer (homogenization time 10s / time, interval 30s, 4 times in a row, in ice water). The prepared 10% homogenate was centrifuged at about 2000r / min in a low-temperature low-speed centrifuge for 10-15 minutes, and the supernatant was taken. The cAMP and cGMP levels in the ciliary bodies of the experimental rabbits were determined using ELISA kits. The above kits were purchased from Wuhan Gilead Biotechnology Co., Ltd.

[0114] Data processing: SPSS statistical software was used for data analysis. All measured data are expressed as mean ± standard deviation. Statistical analysis was performed using the t-test, with P < 0.05 considered statistically significant.

[0115] The experimental results are shown in Table 3. Compared with the blank group, the cAMP content in the ciliary body tissue of the model group was significantly reduced, while the cGMP content was significantly increased, indicating that the model was successfully established.

[0116] Compared with the model group, the cAMP content in the ciliary tissue of the Example 1-3 and Comparative Example 1-8 groups fed with the corresponding composition for 4 weeks was significantly increased, and the cGMP content was significantly decreased, indicating that the compositions prepared in the Examples and Comparative Examples have a regulatory effect on the abnormal cAMP and cGMP levels in the ciliary tissue of the eye caused by the animal model of visual fatigue.

[0117] Among them, the effects of Examples 1-3 and Comparative Examples 1-7 are better than those of the positive control group, while the effect of Comparative Example 8 is worse than that of the positive control group, indicating that the mixed fermentation step plays an important role in the release of the active ingredients of the Chinese medicine composition, making the prepared Chinese medicine fermentation composition more effective. Comparative Examples 1-2 lack one of the strains of Lactobacillus rhamnosus and Saccharomyces cerevisiae, respectively. Compared with the Chinese medicine fermentation compositions prepared in the above comparative examples and Examples 1-3, the improvement effect on the visual fatigue animal model is significantly weakened; indicating that the composition of different species in the composite bacteria directly affects the fermentation results, and the experiment shows that Lactobacillus rhamnosus, Saccharomyces cerevisiae and Aspergillus niger can promote each other in fermentation, influence each other and produce a synergistic effect, which can effectively improve the therapeutic effect of the composition. Furthermore, the inoculation ratio of Lactobacillus rhamnosus, Saccharomyces cerevisiae and Aspergillus niger in Comparative Examples 3-4 exceeds a certain range. Compared with the Chinese medicinal fermentation compositions prepared in the above comparative examples and Examples 1-3, the improvement effect on the visual fatigue animal model is also significantly weakened, indicating that different ratios of the same strains will also affect the fermentation results to a certain extent, and have a certain impact on the therapeutic effect of the Chinese medicinal fermentation composition; and the inoculation ratio of Lactobacillus rhamnosus, Saccharomyces cerevisiae and Aspergillus niger is 1.5:1:1.2, which has the best fermentation effect. Finally, different strains of Lactobacillus rhamnosus, Saccharomyces cerevisiae and Aspergillus niger were replaced in Comparative Examples 5-7, respectively. The experimental results of the above comparative examples compared with the embodiments show that even if the same strains are used, the differences in the properties of different strains will affect the effect of the traditional Chinese medicine fermentation composition; further, different Aspergillus niger strains were replaced in Comparative Example 7, and its effect was worse than that of Comparative Examples 5-6, indicating that different Aspergillus niger strains have a greater impact on the fermentation effect than Lactobacillus rhamnosus and Saccharomyces cerevisiae; further, different strains of Lactobacillus rhamnosus and Saccharomyces cerevisiae were replaced in Comparative Examples 5 and 6, respectively, which, compared with the experimental results of the embodiments, verified that the Lactobacillus rhamnosus and Saccharomyces cerevisiae strains selected by the present invention were obtained by the inventor through a large number of experimental screenings in the early stage and have the best synergistic effect with the Aspergillus niger strain.

[0118] In summary, the present invention uses a mixed bacteria fermentation of Lactobacillus rhamnosus, Saccharomyces cerevisiae and Aspergillus niger to produce a Chinese medicinal ingredient after enzymatic hydrolysis. The above bacteria have a good synergistic effect during the fermentation process, so that the prepared Chinese medicinal fermentation composition is safe and non-irritating, and has a significant regulatory effect on the abnormal cAMP and cGMP levels in the ciliary body tissue caused by the visual fatigue animal model. It can solve the problems of long treatment course and slow effect in the existing Chinese medicine treatment of visual fatigue.

[0119] Table 3 Efficacy verification results

[0120] Group cAMP (nmol / L) cGMP (nmol / L) Blank control group 140.80±12.12 31.05±4.76 Model Group 53.91±5.05* 144.93±15.18* Positive control group <![CDATA[71.89±9.13 ▲ ]]> <![CDATA[48.21±6.46 ▲ ]]> Example 1 <![CDATA[98.96±10.56 ▲# ]]> <![CDATA[37.23±5.39 ▲# ]]> Example 2 <![CDATA[100.57±8.83 ▲# ]]> <![CDATA[35.77±4.25 ▲# ]]> Example 3 <![CDATA[97.78±9.14 ▲# ]]> <![CDATA[38.59±6.57 ▲# ]]> Comparative Example 1 <![CDATA[75.03±8.15 ▲ ]]> <![CDATA[50.11±7.53 ▲ ]]> Comparative Example 2 <![CDATA[69.97±9.27 ▲ ]]> <![CDATA[55.67±6.47 ▲# ]]> Comparative Example 3 <![CDATA[84.12±10.38 ▲# ]]> <![CDATA[46.91±5.61 ▲ ]]> Comparative Example 4 <![CDATA[86.87±8.01 ▲# ]]> <![CDATA[45.43±6.84 ▲ ]]> Comparative Example 5 <![CDATA[92.18±9.33 ▲# ]]> <![CDATA[40.26±7.18 ▲# ]]> Comparative Example 6 <![CDATA[91.79±11.21 ▲# ]]> <![CDATA[41.92±5.39 ▲# ]]> Comparative Example 7 <![CDATA[89.10±10.08 ▲# ]]> <![CDATA[43.79±6.88 ▲# ]]> Comparative Example 8 <![CDATA[62.38±7.76 ▲# ]]> <![CDATA[60.24±8.97 ▲# ]]>

[0121] Note: Compared with the blank group: P < 0.05 is indicated by "*"; compared with the model group: P < 0.05 is indicated by "▲ " Compared with the positive control group: P < 0.05 is indicated by " # "express.

[0122] Application Example 1

[0123] The preparation method of the Chinese medicinal fermentation composition of this application example is consistent with that of Example 2. The massage cream is prepared from the following raw materials in percentage by weight: 14.62% Chinese medicinal fermentation composition, 1.5% steareth-2, 2.0% steareth-21, 4.0% glyceryl stearate, 5.0% cetearyl alcohol, 5.0% dioctyl carbonate, 3.0% isopropyl myristate, 2.0% dimethicone, 0.05% bisabolol, 47.1 % Water, 5.0% 1,3 Propanediol, 0.3% Xanthan Gum, 1.0% Hydroxypropyl Starch Phosphate, 0.1% Disodium EDTA, 0.5% Betaine, 0.1% Allantoin, 0.2% Panthenol, 0.5% Titanium Dioxide, 5.0% Glycerin, 2.0% Butylene Glycol, 0.03% Tetrahydropiperine, 0.1% Methylisothiazolinone or Iodopropynyl Butylcarbamate, 0.1% Fragrance, 0.8% Phenoxyethanol or Ethylhexylglycerin.

[0124] The visual function of the massage cream prepared in Example 1 was tested, and the test was carried out with reference to the "Health Food Functional Test and Evaluation Method (2023 Edition)".

[0125] Inclusion criteria: Gender is not limited, age is 18-65 years old; those who use their eyes for a long time and are prone to visual fatigue. Exclusion criteria: patients with internal and external eye diseases such as corneal, lens, vitreous, and fundus diseases; patients with cardiovascular, cerebrovascular, liver, kidney, and hematopoietic system diseases; pregnant or lactating women, and patients with allergies; those who take items related to the tested function in the short term, which may affect the judgment of the results; those who can cooperate with the experiment; those who are deemed unsuitable for participation by the researchers. The subjects were randomly divided into an experimental group and a control group of 63 people each. The experimental group used the massage cream prepared in Application Example 1, and the control group used the massage cream prepared using the Chinese medicine composition prepared in Comparative Example 8.

[0126] 1) Usage: After cleansing the skin, first cover both eyes with a layer of gauze measuring 5 x 6 cm. Then, apply an appropriate amount of the cream to the entire eye area and gently massage. Then, cover with a layer of plastic wrap, and then cover with two layers of gauze measuring 5 x 6 cm. Leave on for 15 minutes, then remove the plastic wrap and gauze. Wipe off any remaining cream with a damp cotton ball or towel.

[0127] 2) Improvement of discomfort symptoms: Symptom inquiry included 8 symptoms of general discomfort related to eye swelling, eye pain, photophobia, blurred vision, dry eyes, foreign body sensation, tearing, and visual fatigue. Symptom improvement was considered if 3 of them improved, and no other symptoms worsened. Symptom improvement efficacy (%) = number of cases with improved symptoms / total number of cases × 100%;

[0128] 3) Determination of the persistence of clear vision: The percentage of the clear vision time to the fixation time is called the persistence of clear vision; Draw a three-dimensional "pin" shaped cube diagram on the checklist, with each side of the cube being 1 cm long and the local illumination being 120 LX. The distance between the checklist and the eyes should be kept at an appropriate distance without movement according to the viewing habit of the subject. It is stipulated that when the subject sees the "pin" shaped image, it is considered clear vision, and when it is an inverted "pin", it is considered unclear vision. The measurement time is 3 minutes, and the average value is taken after measuring twice. Persistence of clear vision = (Clear vision time / Total fixation time) × 100%;

[0129] 4) Statistical processing of experimental results: The experimental results are shown in Table 4-5. The experimental data are statistically processed using Microsoft Excel 2003, and the results are expressed as mean ± standard deviation. Statistical analysis uses the t-test, and P < 0.05 is considered statistically significant;

[0130] 5) Results: During the trial process of the patients, no symptoms such as skin itching, stinging, and swelling occurred. The experimental results are shown in Table 4-5.

[0131] Table 4 Improvement results of discomfort symptoms

[0132]

[0133] Table 5 Changes in the persistence of clear vision before and after the trial

[0134] Group Before the test After the test Application Example 1 56.49±8.76 <![CDATA[71.34±9.85* # ]]> control group 57.01±7.86 60.37±7.62 <​​​​​​​​​​​The above description is merely a preferred embodiment of the present invention and does not constitute any form of limitation to the present invention. Although the present invention has been disclosed as above in terms of a preferred embodiment, it is not intended to limit the present invention. Any person skilled in the art can, without departing from the scope of the technical solution of the present invention, make some changes or modifications to equivalent embodiments using the technical contents disclosed above. However, any brief modifications, equivalent changes and modifications made to the above embodiments based on the technical essence of the present invention without departing from the content of the technical solution of the present invention are still within the scope of the technical solution of the present invention.

Claims

1. A method for preparing a traditional Chinese medicine fermentation composition for relieving visual fatigue, characterized in that: It consists of the following steps: S1. Weigh each raw material in the Chinese medicine composition according to parts by weight, grind, and sieve to obtain a mixed powder; add sterile deionized water to the mixed powder, stir evenly, and obtain a mixed solution I; S2, adding pectinase and cellulase to the mixed solution I, and performing ultrasound-assisted enzymolysis; after the enzymolysis is completed, heating the solution to inactivate the enzymes to obtain a mixed solution II; S3, respectively activating Lactobacillus rhamnosus, Saccharomyces cerevisiae, and Aspergillus niger with liquid culture medium, and inoculating the activated Lactobacillus rhamnosus, Saccharomyces cerevisiae, and Aspergillus niger into mixed solution II for mixed fermentation; S4, after the fermentation is completed, the filtrate is filtered and concentrated under reduced pressure to obtain a concentrate; the concentrate is sterilized at high temperature to obtain the Chinese medicine fermentation composition; Among them, the Lactobacillus rhamnosus preservation number is CGMCC 1.3724; the Saccharomyces cerevisiae preservation number is CGMCC2.1527; the Aspergillus niger preservation number is CGMCC 3.316; the volume ratio of Lactobacillus rhamnosus, Saccharomyces cerevisiae and Aspergillus niger is 1-2:1:1-1.5; The traditional Chinese medicine composition is composed of the following raw materials in parts by weight: 3-15 parts of wolfberry, 1-10 parts of chrysanthemum, 6-20 parts of Xing'an Cimicifuga, 1-10 parts of Ligustrum lucidum, 5-16 parts of Ligustrum lucidum, 5-16 parts of safflower, 1-10 parts of coptis chinensis, 1-10 parts of equisetum, 1-10 parts of gardenia, 1-10 parts of tribulus terrestris, 5-16 parts of dodder seed, 6-18 parts of cassia seed, 1-10 parts of buddlejac, 1-10 parts of white peony root, and 1-10 parts of mulberry leaf.

2. The method for preparing the Chinese medicinal fermentation composition according to claim 1, wherein The traditional Chinese medicine composition is composed of the following raw materials in parts by weight: 10 parts of wolfberry, 4 parts of chrysanthemum, 13 parts of Xing'an Cimicifuga, 5 parts of Ligustrum lucidum, 10 parts of Ligustrum lucidum, 10 parts of safflower, 4 parts of coptis chinensis, 5 parts of equisetum, 5 parts of gardenia, 5 parts of tribulus terrestris, 11 parts of dodder seed, 12 parts of cassia seed, 4 parts of buddlejac, 4 parts of white peony root, and 4 parts of mulberry leaf.

3. The method for preparing the Chinese medicinal fermentation composition according to claim 1, wherein In the step S1, the mesh number of the sieve is 200-300 mesh, and the material-liquid ratio of the mixed powder of the traditional Chinese medicine composition to sterile deionized water is 1:5-7.

4. The method for preparing the Chinese medicinal fermentation composition according to claim 1, wherein In step S2, the mass ratio of pectinase to cellulase is 2:1, the total enzyme amount is 3-5wt% of the mass of the mixed solution I, the enzymatic hydrolysis temperature is 45-55°C, the pH value is 4.5-5.5, the ultrasonic frequency is 25-35kHz, and the time is 40-50min; the enzyme inactivation treatment temperature is 90-100°C, and the enzyme inactivation treatment time is 20-30min.

5. The method for preparing the Chinese medicinal fermentation composition according to claim 1, wherein In step S3, the total amount of Lactobacillus rhamnosus, Saccharomyces cerevisiae and Aspergillus niger added is 5-7% of the volume of the mixed solution II.

6. The method for preparing the Chinese medicinal fermentation composition according to claim 1, wherein In step S3, the volume ratio of Lactobacillus rhamnosus, Saccharomyces cerevisiae and Aspergillus niger is 1.5:1:1.

2.

7. The method for preparing the Chinese medicinal fermentation composition according to claim 1, wherein In step S3, the fermentation temperature is 30-34° C., the fermentation pH is 5.0-6.0, the sterile air ventilation volume is 5 L / min, the rotation speed is 200-250 rpm, and the fermentation time is 48-72 h.

8. The method for preparing the Chinese medicinal fermentation composition according to claim 1, wherein In the step S4, the filtrate is concentrated under reduced pressure to 80% of the mass of the original filtrate, the temperature of the concentrated under reduced pressure is 40-50° C., the pressure of the concentrated under reduced pressure is -0.06 to -0.08 MPa; the sterilization temperature is 121° C., and the sterilization time is 20-30 min.

9. A massage cream for relieving visual fatigue, characterized in that: The massage cream comprises a traditional Chinese medicine fermentation composition prepared by the preparation method according to any one of claims 1 to 8.

10. The massage cream for relieving visual fatigue according to claim 9, characterized in that: The massage cream comprises the following raw materials in percentage by weight: 5.0-20.0% of a traditional Chinese medicine fermentation composition, 6.5-8.5% of an emulsifier, 3.0-8.0% of an excipient, 8.0-13.0% of a softener, 0.18-0.68% of a conditioner, 0.3-2.3% of a suspending agent, 0.1-1.0% of a covering agent, 10.5-14.5% of a moisturizing agent, and 0.05-0.15% of a chelating agent.

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