Anti-aging and anti-oxidation composition as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of composition processing, mainly relates to a composition and a preparation method thereof, and particularly relates to an anti-aging and anti-oxidation composition and a preparation method thereof. The ginseng, the astragalus membranaceus, the gynostemma pentaphyllum, the semen cassiae, the lotus leaves and the hawthorns are used as main raw materials, and the composition is prepared through synergistic fermentation of the bifidobacterium bifidum CICC-6170 and the lactobacillus acidophilus CICC-6095 to improve the performance and has the effects of resisting aging, resisting oxidation and controlling in-vivo fat.

Description

Technical Field

[0001] The present invention belongs to the technical field of composition processing, and mainly relates to a composition and a preparation method thereof, and specifically relates to an anti-aging and antioxidant composition and a preparation method thereof. Background Art

[0002] Aging is an irreversible and inevitable process. With the increase of age, under the combined effects of internal and external environments, biological molecules, cells, tissues and organs in the human body will undergo harmful and irreparable changes, which may induce a variety of diseases, such as neurodegenerative diseases, tumors and cardiovascular diseases. The most classic is the "free radical theory", which believes that oxidative stress is the primary factor driving aging, and the use of free radical inhibitors and antioxidants can delay aging.

[0003] With the intensification of the global aging trend and the improvement of people's health awareness, the market demand for anti-aging and antioxidant products is growing. At present, there are many kinds of products on the market, but most of them rely on chemical synthetic ingredients or single nutrient supplements, lacking comprehensive and natural anti-aging and antioxidant effects. At the same time, many products fail to fully utilize the advantages of traditional Chinese medicinal materials, resulting in limited effects in improving human functions and enhancing immunity.

[0004] In view of the above-mentioned deficiencies, the present invention proposes an anti-aging and antioxidant composition, which is characterized in that it comprehensively adopts substances with medicinal and edible properties as main raw materials, and combines modern scientific and technological means for processing and extraction, aiming to provide a natural, safe and comprehensive anti-aging and antioxidant composition. Summary of the invention

[0005] In view of this, the present invention provides an anti-aging and antioxidant composition and a preparation method thereof, which uses ginseng, astragalus, gynostemma pentaphyllum, cassia seed, lotus leaf and hawthorn as main raw materials, and obtains a composition by improving performance through synergistic fermentation of Bifidobacterium bifidum CICC-6170 and Lactobacillus acidophilus CICC-6095. The composition has the effects of anti-aging, antioxidant and controlling body fat.

[0006] In a first aspect, the present invention provides a method for preparing a composition, the specific steps of which are as follows: S1. Pretreatment of raw materials: weigh 10-15g ginseng, 8-12g astragalus, 5-8g gynostemma pentaphyllum, 3-5g hawthorn, 5-8g cassia seed, 3-5g lotus leaf, wash and slice, dry and crush; S2. Preparation of Gynostemma pentaphyllum extract: add 8-10 times of ethanol to the Gynostemma pentaphyllum prepared in step S1, stir evenly, soak for 30 minutes, stir and reflux at 60° C. for 1.5 hours, reflux and extract three times, combine the extracts and concentrate under reduced pressure to completely remove ethanol, to obtain a Gynostemma pentaphyllum extract.

[0007] S3. Activation of Bifidobacterium bifidum: strain: Bifidobacterium bifidum, with a collection number of CICC-6170, provided by China Industrial Microbiological Culture Collection Management Center. The strain was inoculated on RCM agar medium and cultured in an oven at 37° C. until milky white colonies were formed; S4. Activation of Lactobacillus acidophilus: strain: Lactobacillus acidophilus, collection number CICC-6095, provided by China Industrial Microbiological Culture Collection Management Center, inoculate the strain on MRS agar medium, place in an oven at 37° C. for culture, and culture until white colonies are formed; S5. Preparation of Bifidobacterium bifidum bacterial solution: The Bifidobacterium bifidum activated in step S3 was transferred to a shake flask containing 500-600 mL of RCM liquid medium and fermented for 6-8 hours to obtain a concentration of 3.0-3.5×10 7 cfu / mL of Bifidobacterium bifidum bacterial solution; S6, preparation of fermented bacterial liquid of Lactobacillus acidophilus: the Lactobacillus acidophilus activated in step S4 was transferred to a shake flask containing 500-600 mL of MRS liquid medium and fermented for 6-8 h to obtain a liquid liquid with a concentration of 2.5-3.0×10 7 cfu / mL of Lactobacillus acidophilus liquid; S7, preparation of fermented product: stir ginseng, astragalus, hawthorn, lotus leaf, cassia seed and the extract of Gynostemma pentaphyllum prepared in step S1 and place in a beaker containing 20-25 mL of Bifidobacterium bifidum CICC-6170 culture liquid prepared in step S5 and 20-25 mL of Lactobacillus acidophilus CICC-6095 culture liquid prepared in step S6 and 20-25 mL of basal medium, and ferment anaerobically at 37°C for 20-22h. After the fermentation, inactivate Bifidobacterium bifidum and Lactobacillus acidophilus in the obtained fermented liquid with 3% H2O2 solution; S8. Preparation of the composition: add 8-10 times of ethanol to the fermented product prepared in step S7, stir evenly, extract under reflux at 60°C for 1.5 h, repeat the extraction 3 times, combine the extracts, concentrate under reduced pressure until the ethanol is completely removed, and then put them into a centrifuge and centrifuge at 3000 r / min for 15 min, take the supernatant, and you can get the composition of the present invention.

[0008] In a second aspect, the composition prepared by the above method has the effects of anti-aging, anti-oxidation and controlling body fat.

[0009] Preferably, the particle size of the crushed particles in step S1 of the present invention is 80-100 mesh particles.

[0010] Preferably, the ethanol added in step S2 and step S8 of the present invention is ethanol with a volume fraction of 60-70%.

[0011] Preferably: the RCM agar medium used in step S3 of the present invention comprises: 5 g peptone, 10 g beef extract powder, 3 g yeast extract powder, 5 g glucose, 1 g soluble starch, 5 g sodium chloride, 3 g sodium acetate, 0.5 g L-cysteine ​​hydrochloride, 15 g agar, and 1000 mL distilled water.

[0012] Preferably: the components of the MRS agar medium used in step S4 of the present invention are: 10 g casein peptone, 10 g beef extract powder, 5 g yeast extract powder, 20 g glucose, 1 g Tween, 5 g sodium acetate, 2 g triammonium citrate, 2 g dipotassium hydrogen phosphate, 0.2 g magnesium sulfate heptahydrate, 0.05 g manganese sulfate monohydrate, 15 g agar, and 1000 mL distilled water.

[0013] Preferably, the basic culture medium used in step S7 of the present invention comprises: 10 g of peptone, 10 g of beef extract powder, 4 g of yeast extract powder, 10 g of glucose, and 1000 mL of distilled water.

[0014] Preferably: the composition of the present invention is used to prepare anti-aging, anti-oxidation and body fat control products.

[0015] The present invention has the following beneficial effects: 1. The ginseng fermentation liquid and the astragalus fermentation liquid in the present invention have good antioxidant capacity under the synergistic effect.

[0016] 2. In the present invention, under fermentation conditions, the extracts of ginseng, astragalus and gynostemma pentaphyllum work synergistically to have anti-aging effects.

[0017] 3. The hawthorn fermentation liquid and the astragalus fermentation liquid in the present invention work synergistically to control body fat.

[0018] 4. The composition prepared by the present invention contains active substances such as saponin and polysaccharide, which can enhance immunity and promote cellular immunity and humoral immunity.

[0019] 5. The present invention improves the taste and flavor of the raw materials through the fermentation process, making the prepared composition more easily accepted and liked by consumers. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] In order to more clearly illustrate the technical solutions in the embodiments of the present invention, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative labor.

[0021] Figure 1 It is a comparison chart of the antioxidant effects of Example 1 and Comparative Examples 1-2.

[0022] Figure 2 It is a comparison chart of the fat index, TC and TG concentrations in serum of mice in Example 3 and Comparative Examples 12-15. DETAILED DESCRIPTION

[0023] The following describes the embodiments of the present invention by specific examples, and those skilled in the art can easily understand other advantages and effects of the present invention from the contents disclosed in this specification. The present invention can also be implemented or applied through other different specific embodiments, and the details in this specification can also be modified or changed in various ways based on different viewpoints and applications without departing from the spirit of the present invention. It should be noted that the following embodiments and features in the embodiments can be combined with each other without conflict.

[0024] Ginseng, astragalus, gynostemma pentaphyllum, cassia seed, lotus leaf and hawthorn were all provided by Bozhou Yuanshengtang Pharmaceutical Co., Ltd.

[0025] Example 1

[0026] This example demonstrates that the ginseng fermentation broth and the astragalus fermentation broth have good antioxidant capacity under the synergistic effect.

[0027] This embodiment provides a method for preparing a composition, and the specific steps are as follows: S1. Pretreatment of raw materials: weigh 10g ginseng, 8g astragalus, 5g gynostemma pentaphyllum, 3g hawthorn, 5g cassia seed, and 3g lotus leaf on a balance, wash and slice, dry and crush into 80 mesh particles; S2. Preparation of Gynostemma pentaphyllum extract: add 8 times 60% ethanol to the Gynostemma pentaphyllum prepared in step S1, stir evenly, soak for 30 minutes, stir and reflux at 60°C for 1.5 hours, reflux and extract three times, combine the extracts and concentrate under reduced pressure to completely remove ethanol, to obtain a Gynostemma pentaphyllum extract.

[0028] S3. Activation of Bifidobacterium bifidum: strain: Bifidobacterium bifidum, with a collection number of CICC-6170, provided by China Industrial Microbiological Culture Collection Management Center. The strain was inoculated on RCM agar medium and cultured in an oven at 37° C. until milky white colonies were formed; S4. Activation of Lactobacillus acidophilus: strain: Lactobacillus acidophilus, collection number CICC-6095, provided by China Industrial Microbiological Culture Collection Management Center, inoculate the strain on MRS agar medium, place in an oven at 37° C. for culture, and culture until white colonies are formed; S5. Preparation of Bifidobacterium bifidum bacterial solution: The Bifidobacterium bifidum activated in step S3 was transferred to a 500 mL RCM liquid medium shake flask and fermented for 6 h to obtain a concentration of 3.0×10 7 cfu / mL of Bifidobacterium bifidum bacterial solution; S6, preparation of Lactobacillus acidophilus bacterial liquid: The Lactobacillus acidophilus activated in step S4 was transferred to a 500 mL shake flask of MRS liquid culture medium and fermented for 6 h to obtain a concentration of 2.5×10 7 cfu / mL of Lactobacillus acidophilus liquid; S7, preparation of fermented product: ginseng, astragalus, hawthorn, lotus leaf, cassia seed and the extract of Gynostemma pentaphyllum prepared in step S1 are stirred evenly and placed in a beaker containing 20 mL of the bacterial solution of Bifidobacterium bifidum CICC-6170 prepared in step S5, 20 mL of the bacterial solution of Lactobacillus acidophilus CICC-6095 prepared in step S6 and 20 mL of the basal medium, and anaerobically fermented at 37° C. for 20 h. After the fermentation is completed, the Bifidobacterium bifidum and Lactobacillus acidophilus in the obtained fermented liquid are inactivated with a 3% H2O2 solution, and a fermented product is obtained after the inactivation is completed; S8. Preparation of the composition: add 8 times of ethanol to the fermented product prepared in step S7, reflux extract at 60°C for 1.5 h, repeat the extraction 3 times, combine the extracts and concentrate under reduced pressure until the ethanol is completely removed, then put them into a centrifuge and centrifuge at 3000 r / min for 15 min, and take the supernatant to obtain the composition.

[0029] Comparative Example 1: Ginseng is not added in step S7, and the remaining steps are the same as those in Example 1.

[0030] Comparative Example 2: In step S7, no astragalus was added, and the remaining steps were the same as those in Example 1.

[0031] Determination of DPPH free radical scavenging rate: First, mix 200 uL of the sample with 200 uL of 0.2 mmol / L DPPH anhydrous ethanol solution, and then place it in a dark place without light at room temperature for 30 minutes. After the reaction is completed, use an enzyme reader to measure the absorbance of the mixture at 517nm. The blank control is a mixture obtained by mixing 200uL of distilled water and 200uLDPPH-anhydrous ethanol solution; the control group is a mixture obtained by mixing 200uL of sample solution and 200uL of anhydrous ethanol. Vitamin C is used as the positive control group. According to the absorbance data obtained, the DPPH free radical scavenging rate (R1, %) is calculated using the relevant formula. The results are shown in Figure 1 shown.

[0032] R1 / %=[1-(A 样 -A 对 ) / A 空 ]×100, where: A 样 , A 对 , A 空 are the absorbance of the sample group, control group, and blank group, respectively.

[0033] (2) Determination of OH radical scavenging rate: First, 200 uL of sample was mixed with 200 uL of 9 mmol / L ferrous sulfate solution, 200 uL of 9 mmol / L salicylic acid ethanol solution, and 200 uL of 9 mmol / L hydrogen peroxide solution. The mixture was placed in a water bath at 37°C for 30 min. After standing for 20 min, the absorbance of the mixture at 510 nm was measured using an ELISA reader. The blank group was replaced with distilled water, the control group was a mixed solution of 200 uL sample solution, 400 uL distilled water, and 200 uL anhydrous ethanol. Vitamin C was used as a positive control. Based on the absorbance data obtained, the OH radical scavenging rate (R2, %) was calculated using the relevant formula.

[0034] R2 / %=[1-(A 样 -A 对 ) / A 空 ]×100, where: A 样 , A 对 , A 空 are the absorbance of the sample group, control group, and blank group, respectively. Figure 1 As shown: Figure 1It is a comparison chart of the antioxidant effects of Example 1 and Comparative Examples 1-2. It can be seen from the results that in the DPPH radical scavenging rate determination experiment, the elimination rates of DPPH radicals in the compositions of Example 1 and Comparative Examples 1-2 were 90.25%, 72.28% and 74.50%, respectively, among which the elimination rate of DPPH radicals in the composition of Example 1 was the strongest; in the ·OH radical scavenging rate determination experiment, the elimination rates of ·OH radicals in the compositions of Example 1 and Comparative Examples 1-2 were 86.36%, 67.37% and 69.95%, respectively, among which the elimination rate of ·OH radicals in the composition of Example 1 was the strongest. It can be seen that compared with Comparative Examples 1 and 2, the elimination rates of DPPH radicals and ·OH radicals in Example 1 were the strongest, and the antioxidant effect was the best, which confirmed that the ginseng fermented liquid and the astragalus fermented liquid had good antioxidant capacity under the synergistic effect.

[0035] Example 2

[0036] This example demonstrates that under fermentation conditions, the extracts of ginseng, astragalus and gynostemma pentaphyllum have anti-aging effects under synergistic action.

[0037] This embodiment provides a method for preparing a composition, and the specific steps are as follows: S1. Pretreatment of raw materials: weigh 12g ginseng, 10g astragalus, 6g gynostemma pentaphyllum, 4g hawthorn, 6g cassia seed, 4g lotus leaf on a balance, wash and slice, dry and crush into 80 mesh particles; S2. Preparation of Gynostemma pentaphyllum extract: add 9 times 65% ethanol to the Gynostemma pentaphyllum prepared in step S1, stir evenly, soak for 30 minutes, stir and reflux at 60°C for 1.5 hours, reflux and extract three times, combine the extracts and concentrate under reduced pressure to completely remove ethanol, to obtain a Gynostemma pentaphyllum extract.

[0038] S3. Activation of Bifidobacterium bifidum: strain: Bifidobacterium bifidum, with a collection number of CICC-6170, provided by China Industrial Microbiological Culture Collection Management Center. The strain was inoculated on RCM agar medium and cultured in an oven at 37° C. until milky white colonies were formed; S4. Activation of Lactobacillus acidophilus: strain: Lactobacillus acidophilus, collection number CICC-6095, provided by China Industrial Microbiological Culture Collection Management Center, inoculate the strain on MRS agar medium, place in an oven at 37° C. for culture, and culture until white colonies are formed; S5. Preparation of Bifidobacterium bifidum bacterial solution: The Bifidobacterium bifidum activated in step S3 was transferred to a 550 mL RCM liquid medium shake flask and fermented for 7 hours to obtain a concentration of 3.2×10 7cfu / mL of Bifidobacterium bifidum bacterial solution; S6, preparation of Lactobacillus acidophilus bacterial liquid: The Lactobacillus acidophilus activated in step S4 was transferred to a 550 mL shake flask of MRS liquid culture medium and fermented for 7 h to obtain a concentration of 2.7×10 7 cfu / mL of Lactobacillus acidophilus liquid; S7, preparation of fermented product: ginseng, astragalus, hawthorn, lotus leaf, cassia seed and the extract of Gynostemma pentaphyllum prepared in step S1 are stirred evenly and placed in a beaker containing 22 mL of the bacterial solution of Bifidobacterium bifidum CICC-6170 prepared in step S5, 22 mL of the bacterial solution of Lactobacillus acidophilus CICC-6095 prepared in step S6 and 22 mL of the basal medium, and anaerobically fermented at 37° C. for 21 h. After the fermentation is completed, the Bifidobacterium bifidum and Lactobacillus acidophilus in the obtained fermented liquid are inactivated with a 3% H2O2 solution, and a fermented product is obtained after the inactivation is completed; S8. Preparation of the composition: Add 9 times of ethanol to the fermented product prepared in step S7, reflux extraction at 60°C for 1.5 h, repeat the extraction 3 times, combine the extracts, and concentrate under reduced pressure until the ethanol is completely removed, then put them into a centrifuge and centrifuge at 3000 r / min for 15 min, and take the supernatant to obtain the composition.

[0039] Comparative Example 3: Except that ginseng is not added in step S7, the remaining steps are the same as those in Example 2.

[0040] Comparative Example 4: Except that Astragalus is not added in step S7, the remaining steps are the same as those in Example 2.

[0041] Comparative Example 5: Except that no Gynostemma pentaphyllum extract is added in step S7, the remaining steps are the same as those in Example 2.

[0042] Comparative Example 6: Except that Bifidobacterium bifidum CICC-6170 and Lactobacillus acidophilus CICC-6095 were not added in step S7 and fermentation was carried out directly, the remaining steps were the same as those in Example 2.

[0043] Comparative Example 7: Except that Bifidobacterium bifidum CICC-6170 was not added in step S7, the remaining steps were the same as those in Example 2.

[0044] Comparative Example 8: Except that Lactobacillus acidophilus CICC-6095 is not added in step S7, the remaining steps are the same as those in Example 2.

[0045] Comparative Example 9: Except that Bifidobacterium bifidum CICC-6170 was replaced by Bifidobacterium bifidum CICC-6171 (purchased from China Industrial Microbiological Culture Collection Center) in step S7, the remaining steps were the same as those in Example 2.

[0046] Comparative Example 10: Except that Lactobacillus acidophilus CICC-6095 was replaced by Lactobacillus acidophilus CICC-20540 (purchased from China Industrial Microbiological Culture Collection Center) in step S7, the remaining steps were the same as those in Example 2.

[0047] Animal experiment: KM mice were provided by Beijing Weitong Lihua Experimental Animal Technology Co., Ltd., weighing 20±2g and with SPF grade hygiene. 120 mice were randomly placed in separate cages at room temperature, while ensuring that they had free access to food and water. The experimental process adopted the "3R" (replacement, reduction, optimization) experimental design principle. During the entire experiment, a comfortable environment was created for the animals as much as possible, and standardized operating procedures were adopted to minimize their pain.

[0048] Grouping and administration: After the mice were fully adapted to the environment, 120 mice were randomly divided into 12 groups, 10 mice in each group, namely, model group, negative control group, positive control group (piracetam), Example 2 group, and Comparative Examples 3-10 groups. The Example 2 group and Comparative Examples 3-10 groups were intragastrically administered with the corresponding composition, which was diluted to 10% v / v with purified water, and the intragastrically administered dose was 15 mL / kg; the positive control group was intragastrically administered with piracetam, 600 mg / kg; the model group and the negative control group were intragastrically administered with 0.9% NaCl, 15 mL / kg; at the same time, the negative control group was intraperitoneally injected with 0.9% NaCl 100 mg / kg; and the other groups were intraperitoneally injected with 2% D-galactose 140 mg / kg. Continuous administration was maintained for 49 days.

[0049] Determination of the content of SOD, NOS, MDA, and CAT in mouse brain tissue: After 49 days, the mice were killed by cervical dislocation and their brains were quickly removed. The surface of the brain tissue was washed with ice-cold 0.9% NaCl solution to remove blood stains, and then dried with filter paper. Next, the hippocampal tissue was extracted from the brain, and 9 times of 4°C 0.9% NaCl solution was added and mixed in a homogenization tube to make a homogenate. The homogenate was placed in a low-temperature centrifuge at 4°C and centrifuged at 3000r / min for 15min. After centrifugation, the supernatant was collected and the content of SOD, NOS, MDA, and CAT was determined according to the instructions. The results are shown in Table 1. Among them, superoxide dismutase (SOD), catalase (CAT); nitric oxide synthase (NOS), malondialdehyde (MDA) kits were all purchased from Nanjing Jiancheng Bioengineering Institute.

[0050] Table 1 Group SOD (U / mg) NOS (U / mg) CAT (U / mg) MDA (nmol / mg) Negative control group 125.32±4.57 7.98±0.47 16.12±0.96 8.01±0.95 Model Group 87.17±2.88 3.71±0.42 8.69±0.69 18.21±0.68 Positive control group 149±3.72 6.92±0.59 17.99±0.72 10.20±0.71 Example 2 Group 169.65±5.16 7.88±0.46 17.21±1.02 9.21±0.88 Comparative Example 3 101.21±3.67 4.96±0.47 10.69±0.71 16.48±0.73 Comparative Example 4 116.47±3.73 5.01±0.48 11.72±0.73 15.12±0.78 Comparative Example 5 108.68±3.70 4.99±0.47 11.37±0.72 15.95±0.77 Comparative Example 6 96.58±3.02 4.55±0.45 9.68±0.70 16.98±0.74 Comparative Example 7 127.97±3.89 5.23±0.53 12.69±0.88 14.02±0.83 Comparative Example 8 132.68±4.12 5.29±0.55 12.75±0.91 13.74±0.85 Comparative Example 9 130.49±4.01 5.27±0.54 12.73±0.89 13.77±0.84 Comparative Example 10 139.28±4.28 5.33±0.56 12.89±0.92 13.62±0.86 SOD is one of the main antioxidant enzymes in the body. The increase in its content can enhance the free radical scavenging ability and reduce cell damage, thereby having the effect of delaying aging; the increase in NOS content can increase the production of NO, improve learning and memory ability, thereby resisting the decline in physiological functions related to aging; the increase in CAT content helps to protect the body's organs from damage caused by free radicals, thereby reducing oxidative damage and resisting aging; MDA is a lipid peroxidation product produced under the action of free radicals, and the reduction in its content can reduce cell membrane lipid oxidative damage, thereby protecting the cell membrane and resisting aging. Table 1 is the effect of the composition of Example 2 and Comparative Examples 3-10 on the content of SOD, NOS, CAT, and MDA in the brain tissue of aging model mice. As can be seen from the results, compared with the mice in the negative control group, the content of SOD, NOS, and CAT in the brain tissue of the model group mice was significantly reduced, and the content of MDA was significantly increased. Compared with the model group mice, the SOD, NOS, and CAT contents in the brain tissues of the mice in Example 2 and Comparative Examples 3-10 groups were significantly increased, and the MDA content was significantly decreased, among which the SOD, NOS, and CAT contents in the brain tissues of the mice in Example 2 group increased the most, and the MDA content decreased the most.

[0051] In summary, the composition of Example 2 enhances the body's antioxidant capacity and reduces oxidative stress and cell damage by increasing the activity of SOD and CAT and reducing the level of MDA. At the same time, by increasing the activity of NOS, the production of NO is increased, and the learning and memory abilities are improved, and these effects work together to combat the aging process. It is confirmed that the extracts of ginseng, astragalus and gynostemma pentaphyllum have anti-aging effects under the synergistic effect under fermentation conditions.

[0052] Example 3

[0053] This example demonstrates that the synergistic effect of hawthorn fermented liquid, cassia seed fermented liquid and lotus leaf fermented liquid has the effect of controlling body fat.

[0054] This embodiment provides a preparation method, and the specific steps are as follows: S1. Pretreatment of raw materials: weigh 15g ginseng, 12g astragalus, 8g gynostemma pentaphyllum, 5g hawthorn, 8g cassia seed, 5g lotus leaf on a balance, wash and slice, dry and crush into 100 mesh particles; S2. Preparation of Gynostemma pentaphyllum extract: add 10 times 70% ethanol to the Gynostemma pentaphyllum prepared in step S1, stir evenly, soak for 30 minutes, stir and reflux at 60° C. for 1.5 hours, reflux and extract three times, combine the extracts and concentrate under reduced pressure to completely remove ethanol, to obtain a Gynostemma pentaphyllum extract.

[0055] S3. Activation of Bifidobacterium bifidum: strain: Bifidobacterium bifidum, with a collection number of CICC-6170, provided by China Industrial Microbiological Culture Collection Management Center. The strain was inoculated on RCM agar medium and cultured in an oven at 37° C. until milky white colonies were formed; S4. Activation of Lactobacillus acidophilus: strain: Lactobacillus acidophilus, collection number CICC-6095, provided by China Industrial Microbiological Culture Collection Management Center, inoculate the strain on MRS agar medium, place in an oven at 37° C. for culture, and culture until white colonies are formed; S5. Preparation of Bifidobacterium bifidum bacterial solution: The Bifidobacterium bifidum activated in step S3 was transferred to a 600 mL RCM liquid medium shake flask for fermentation for 8 h to obtain a concentration of 3.5×10 7 cfu / mL of Bifidobacterium bifidum bacterial solution; S6, preparation of Lactobacillus acidophilus bacterial liquid: The Lactobacillus acidophilus activated in step S4 was transferred to a 600 mL shake flask of MRS liquid culture medium and fermented for 8 h to obtain a concentration of 3.0×10 7 cfu / mL of Lactobacillus acidophilus liquid; S7, preparation of fermented product: ginseng, astragalus, hawthorn, lotus leaf, cassia seed and the extract of Gynostemma pentaphyllum prepared in step S1 are stirred evenly and placed in a beaker containing 25 mL of the bacterial solution of Bifidobacterium bifidum CICC-6170 prepared in step S5 and 25 mL of the bacterial solution of Lactobacillus acidophilus CICC-6095 prepared in step S6 and 25 mL of the basal medium, and anaerobically fermented at 37°C for 22 hours, and after the fermentation is completed, the Bifidobacterium bifidum and Lactobacillus acidophilus in the obtained fermented liquid are inactivated with 3% H2O2 solution, and the fermented product is obtained after the inactivation is completed; S8. Preparation of a composition: Add 9 times of ethanol to the fermented product prepared in step S7, reflux extraction at 60°C for 1.5 h, repeat the extraction 3 times, combine the extracts, and concentrate under reduced pressure until the ethanol is completely removed, then put them into a centrifuge and centrifuge at 3000 r / min for 15 min, and take the supernatant to obtain a composition.

[0056] Comparative Example 12: Except that hawthorn is not added in step S7, the remaining steps are the same as those in Example 3.

[0057] Comparative Example 13: Except that Cassia seed is not added in step S6, the remaining steps are the same as those in Example 3.

[0058] Comparative Example 14: Except that lotus leaves are not added in step S6, the remaining steps are the same as those in Example 3.

[0059] Comparative Example 15: Except that Bifidobacterium bifidum CICC-6170 and Lactobacillus acidophilus CICC-6095 were not added in step S6, the remaining steps were the same as those in Example 3.

[0060] Experimental animals: 70 SPF-grade ICR mice were provided by Beijing Weitonglihua Experimental Animal Technology Co., Ltd. The weight of the mice was 20±2g. Before the experiment, the mice were adaptively raised in a constant temperature and oxygen supply environment at 25±2℃ for one week.

[0061] Grouping: The mice were randomly divided into 7 groups: blank group, model group, Example 3 group, and Comparative Example 12-15 groups.

[0062] Blank group gavage feed: gavage with the basic feed provided by Haopai Biotechnology Co., Ltd.; Other groups were gavaged with a special high-fat feed, which contained 79g of basal feed, 10g of lard, 10g of egg yolk powder and 1g of cholesterol per 100g of feed; Administration of Example 2 and Comparative Examples 12-15: intragastric administration of the composition, which was diluted with purified water to a concentration of 10% v / v, with an intragastric administration dose of 15 mL / kg; Administration of blank control group and model group: gavage with an equal amount of purified water instead of the composition; Experimental period: continuous gavage for 30 days.

[0063] After 30 days, the mice were fasted for 24 h, and the eyeballs were removed for blood collection. The blood samples were placed in a 37°C constant temperature water bath for 30 min, then centrifuged at 3000 r / min for 15 min, and the supernatant was collected to prepare serum.

[0064] The TC and TG kits provided by Shanghai Jiachu Bioengineering Co., Ltd. were used to measure the concentrations of total cholesterol (TC) and triglycerides (TG) in serum according to the operating specifications. The results are as follows Figure 2 shown.

[0065] After enucleation of the eyeballs, the adipose tissue around the mouse genitals was collected and weighed, and the fat coefficient of each group of mice was calculated.

[0066] Fat coefficient = fat mass around mouse genitals / mouse mass × 100% Figure 2It is a comparison chart of the fat coefficient, serum TC and TG concentration of mice in Example 3 and Comparative Example 12-15 groups. As can be seen from the figure, compared with the blank group, the fat coefficient, serum TC and TG concentration of mice in the model group were significantly increased; compared with the model group, the fat coefficient, serum TC and TG concentration of mice in Comparative Example 12-15 groups were slightly lower, and the fat coefficient, serum TC and TG concentration of mice in Example 3 group were the lowest, indicating that Example 3 has a significant inhibitory effect on fat accumulation and the increase of TC and TG caused by a high-fat diet. It is confirmed that the synergistic effect of hawthorn fermentation liquid, cassia seed fermentation liquid and lotus leaf fermentation liquid has the effect of controlling body fat.

[0067] The above-mentioned embodiments only express the specific implementation methods of the present invention, and the descriptions thereof are relatively specific and detailed, but they cannot be understood as limiting the protection scope of the present invention. It should be pointed out that, for those skilled in the art, without departing from the concept of the present invention, any other changes, modifications, substitutions, combinations, simplifications made without departing from the spirit and principle of the present invention should be equivalent replacement methods and are included in the protection scope of the present invention.

Claims

1. An anti-aging antioxidant composition, characterized in that: The composition is obtained by using medicine-food homologous substances as main raw materials through cooperative fermentation and extraction of composite strains; the medicine-food homologous substances are ginseng, astragalus, gynostemma pentaphyllum, cassia seed, lotus leaf and hawthorn; the composite strains are Bifidobacterium bifidum CICC-6170 and Lactobacillus acidophilus CICC-6095.

2. The anti-aging and antioxidant composition according to claim 1, characterized in that: The main raw materials of the composition include 10-15 parts of ginseng, 8-12 parts of astragalus, 5-8 parts of gynostemma pentaphyllum, 3-5 parts of hawthorn, 5-8 parts of cassia seed and 3-5 parts of lotus leaf by weight.

3. An anti-aging antioxidant composition according to claim 2, characterized in that: The composition has the effect of controlling body fat in addition to its anti-aging and anti-oxidation effects.

4. A method for preparing an anti-aging and antioxidant composition according to any one of claims 1 to 3, characterized in that: The specific preparation method of the composition is as follows: S1. Pretreatment of raw materials: weigh 10-15g ginseng, 8-12g astragalus, 5-8g gynostemma pentaphyllum, 3-5g hawthorn, 5-8g cassia seed, 3-5g lotus leaf, wash and slice, dry and crush; S2, preparation of Gynostemma pentaphyllum extract: add 8-10 times of ethanol to the Gynostemma pentaphyllum prepared in step S1, stir evenly, soak for 30 minutes, stir and reflux at 60°C for 1.5 hours, reflux extraction 3 times, combine the extracts and concentrate under reduced pressure until the ethanol is completely removed to obtain a Gynostemma pentaphyllum extract; S3. Activation of Bifidobacterium bifidum: Bifidobacterium bifidum CICC-6170 was inoculated on RCM agar medium and cultured in an oven at 37° C. until milky white colonies were formed; S4. Activation of Lactobacillus acidophilus: Lactobacillus acidophilus CICC-6095 was inoculated on MRS agar medium and cultured in an oven at 37° C. until white colonies were formed; S5. Preparation of Bifidobacterium bifidum bacterial solution: The Bifidobacterium bifidum activated in step S3 was transferred to a shake flask containing 500-600 mL of RCM liquid medium and fermented for 6-8 h to obtain a concentration of 3.0-3.5×10 7 cfu / mL of Bifidobacterium bifidum bacterial solution; S6, preparation of Lactobacillus acidophilus bacterial liquid: The Lactobacillus acidophilus activated in step S4 was transferred to a shake flask containing 500-600 mL of MRS liquid medium and fermented for 6-8 h to obtain a concentration of 2.5-3.0×10 7 cfu / mL of Lactobacillus acidophilus liquid; S7, preparation of fermented product: stir ginseng, astragalus, hawthorn, lotus leaf, cassia seed and the extract of Gynostemma pentaphyllum prepared in step S1 and place in 20-25 mL of Bifidobacterium bifidum CICC-6170 culture liquid prepared in step S5, 20-25 mL of Lactobacillus acidophilus CICC-6095 culture liquid prepared in step S6 and 20-25 mL of basal medium, and perform anaerobic fermentation at 37°C for 20-22 hours before stopping the fermentation, and inactivate Bifidobacterium bifidum and Lactobacillus acidophilus in the obtained fermented liquid with 3% H2O2 solution; S8. Preparation of the composition: add 8-10 times of ethanol to the fermented product prepared in step S7, stir evenly, reflux extract at 60°C for 1.5 h, repeat the extraction 3 times, combine the extracts, concentrate under reduced pressure until the ethanol is completely removed, and then put them into a centrifuge and centrifuge at 3000 r / min for 15 min, take the supernatant, and obtain the composition.

5. The method for preparing an anti-aging antioxidant composition according to claim 4, characterized in that: The particle size of the crushed particles in step S1 of the present invention is 80-100 mesh particles.

6. The method for preparing an anti-aging antioxidant composition according to claim 4, characterized in that: The ethanol added in step S2 and step S8 of the present invention is ethanol with a volume fraction of 60-70%.

7. The method for preparing an anti-aging antioxidant composition according to claim 4, characterized in that: The RCM agar medium used in step S3 of the present invention comprises: 5 g of peptone, 10 g of beef extract powder, 3 g of yeast extract powder, 5 g of glucose, 1 g of soluble starch, 5 g of sodium chloride, 3 g of sodium acetate, 0.5 g of L-cysteine ​​hydrochloride, 15 g of agar, and 1000 mL of distilled water.

8. The method for preparing an anti-aging antioxidant composition according to claim 4, characterized in that: The components of the MRS agar medium used in step S4 are: 10 g casein peptone, 10 g beef extract powder, 5 g yeast extract powder, 20 g glucose, 1 g Tween, 5 g sodium acetate, 2 g triammonium citrate, 2 g dipotassium hydrogen phosphate, 0.2 g magnesium sulfate heptahydrate, 0.05 g manganese sulfate monohydrate, 15 g agar, and 1000 mL distilled water.

9. The method for preparing an anti-aging antioxidant composition according to claim 4, characterized in that: The basic culture medium used in step S7 comprises: 10 g of peptone, 10 g of beef extract powder, 4 g of yeast extract powder, 10 g of glucose, and 1000 mL of distilled water.

10. An anti-aging and antioxidant composition according to claims 1-3 or a composition obtained by the preparation method of an anti-aging and antioxidant composition according to any one of claims 4-9, which is used for preparing anti-aging, antioxidant and body fat control products.

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