TWI836809B - Ophioglossaceae plant extract for promoting cell growth, cell repair, protecting cells, anti-oxidation and anti-inflammation, and use thereof - Google Patents

Abstract

An Ophioglossaceaeplant extract having biological effects including anti-inflammation, anti-oxidation, cell repair, cell proliferation, or cell protection. The extract comprises at least 10 to 40wt% of flavonoids and 3 to 10wt% of phenols by weight of the extract, wherein the extract is extracted from an Ophioglossaceaeplant comprising Helminthostachys zeylanica.

Description

Ophioglossaceae plant extracts that can promote cell growth, repair cells, protect cells, antioxidant and anti-inflammation and their uses

The present invention relates to the use of a fern plant extract for promoting cell proliferation, repairing, protecting cells, anti-oxidation or anti-inflammation, and in particular to the use of an Ophioglossaceae plant extract.

Ophioglossaceae is a family of ferns with short-lived spores formed by sporangia and enlarged roots. Species belonging to this family usually only grow one frond per year, and only a few species can produce panicles. Generally speaking, the gametophyte of Ophioglossaceae is produced underground. Its spores do not grow under sunlight, and the gametophyte can survive for up to two centuries without forming a sporophyte. The vast majority of Ophioglossaceae plants are terrestrial plants, and only a few grow on tree trunks or in wetland environments.

Medicinal plants of the family Ophioglossum , such as Ophioglossum petiolatum Hook , grow in plains, fieldside meadows, and roadsides in the shade of hillside trees throughout Taiwan, in moist areas. In the field of traditional Chinese medicine, the whole plant parts have been used to clear heat and detoxify, activate blood circulation and remove blood stasis, reduce swelling and relieve pain, etc. Another example is Botrychium officinale , which is endemic to Sichuan. , distributed in Nanchuan, Tianquan, Danba and other places in Sichuan, has the functions of detoxifying, dispersing stagnation, clearing away heat and relieving cough, and is used in the field of traditional Chinese medicine to treat sores, carbuncles, tuberculosis, lung-heat cough and other diseases; and for example, Yin Botrychium virginianum of the subgenus Botrychium is distributed in Zhejiang, Shanxi, Shaanxi, western Hubei, northwestern Yunnan, Europe, North America, Brazil, temperate Asia and other places in China. It has the functions of clearing away heat, detoxifying and reducing swelling. It has the functions of dispersing knots and is specifically used in the field of traditional Chinese medicine to treat lung abscess, conjunctivitis, fatigue, insect bites, scrofula, etc.

Helminthostachys zeylanica , classified in the genus Helminthostachys zeylanica under the family Helminthaceae, is also known as the fallen centipede and is widely used in the field of traditional Chinese medicine. The roots of Helminthostachys zeylanica contain stigmasterol, rock A variety of medicinal ingredients such as fernsterol, dulbitol, and centipedeol have the effects of clearing the lungs and resolving phlegm, dispersing blood stasis and detoxifying. They are commonly used to treat sore throat, swelling and pain from bruises, carbuncles, snake bites, etc., and are used in In modern medicine, it is further used to prevent and treat metabolic syndrome, or to inhibit the growth of cancer cells.

For example, U.S. Patent No. US10272124B2 discloses that the extract obtained by heat reflux extraction of rhizome of Ceylonia ternata with ethanol-water solution has achieved positive results in the treatment of metabolic syndrome. For another example, PCT World Patent No. WO2016097831A4 discloses that the active substance Ugonin K in Ceylonia ternata has an inhibitory effect on the growth of skin cancer cells, providing a new option in the field of anti-cancer.

However, there is still a lack of related technologies or products in the prior art that use the plants of the family Cyperaceae, including Cyperaceae, Pteris cretica, Pteris fasciata, and Cyperus rotundus, to promote skin wound healing, cell growth, repair, anti-oxidation, anti-inflammatory and other cell protection activities.

As we all know, skin is the first barrier to protect an individual, and has functions such as preventing water loss, invasion of pathogenic bacteria, and various environmental damage; modern people are exposed to large amounts of 3C blue light, ultraviolet (UV), ionizing radiation, Drugs and other drugs can easily cause the skin to generate free radicals and continue to accumulate to form oxidative stress, which can have unpredictable destructive effects on intracellular components including DNA, proteins, lipids and other substances, leading to skin aging, lesions and even In the face of the above problems, novel pharmaceuticals, foods or skin care products that can promote wound healing, cell growth, repair, antioxidant, and anti-inflammation are urgently needed to be developed.

To solve the above problems, one object of the present invention is to provide a plant extract of the Ophioglossaceae family, which has the effects of promoting cell growth, repairing cells, protecting cells, anti-oxidation and anti-inflammatory, and comprises at least 10 to 40 wt% of total flavonoid active molecules and 3 to 10 wt% of total polyphenol active molecules based on the total weight of the extract, wherein the extract is extracted from a plant of the Ophioglossaceae family.

The Ophioglossaceae plant extract as described above, wherein the Ophioglossaceae plants include Helminthostachys zeylanica , Botrychium lunaria , and Alpine Ophioglossaceae. Ophioglossum austroasiaticum , Ophioglossum pedunculosum , Ophioglossum petiolatum , Ophioglossum reticulatum or Ophioglossum thermale .

The extract of the plant of the family Ampelopsis as described above, wherein the extraction method of the extract of the plant of the family Ampelopsis includes the following steps: S1) soaking a plant of the family Ampelopsis in an alcohol-water solution to obtain a plant soaking liquid; S2) ultrasonically vibrating the plant soaking liquid at a temperature to obtain a crude plant extract; S3) filtering the crude plant extract to obtain an extract of the plant of the family Ampelopsis, wherein the ultrasonic vibration is a multi-stage continuous ultrasonic vibration, wherein step S1 includes soaking the whole plant of the Ampelopsis plant or any plant part selected from the group consisting of rhizomes, stems, leaves, spikes and a combination of any two or more thereof.

As mentioned above, the multi-stage continuous ultrasonic vibration includes: a first stage, which includes continuously shaking the plant soaking liquid for a first shaking time T ₁ , and then suspending the shaking. The plant soaking liquid reaches a first pause time t ₁ ; a second stage includes continuously shaking the plant soaking liquid for a second shaking time T ₂ , and then pausing to shake the plant soaking liquid for a second pause time t ₂ ; A third stage, which includes continuous shaking of the plant soaking liquid for a third shaking time T ₃ , and then pausing to shake the plant soaking liquid for a third pause time t ₃ ; a fourth stage, which includes continuing to shake the plant The soaking liquid reaches a fourth shaking time T ₄ , and then the plant soaking liquid is paused to shake for a fourth pause time t ₄ ; and a fifth stage includes continuous shaking of the plant soaking liquid for a fifth shaking time T ₅ , Then, the plant soaking solution is paused and shaken for a fifth pause time t ₅ , where T ₁ ＜ T ₂ ＜ T ₃ ＜ T ₄ ＜ T ₅ or T ₁ ＞ T ₂ ＞ T ₃ ＞ T ₄ ＞ T ₅ , and t ₁ ＜t ₂ ＜t ₃ ＜t ₄ ＜t ₅ .

As for the extract of the Echinaceae plant as described above, the extraction method further includes the following steps: S4) Concentrate the Echinaceae plant extract to obtain a concentrated extract; and S5) Concentrate the The extract is freeze-dried to obtain a concentrated extract in the form of a dry powder.

The Ophioglossaceae plant extract as described above, wherein the Ophioglossaceae plant includes rhizomes.

The extract of the plant of the family Cyperaceae as described above, wherein the temperature is 20 to 35° C., the plant soaking liquid comprises 1 to 10 parts by weight of the plant of the family Cyperaceae and 15 to 150 parts by weight of an alcohol aqueous solution, the alcohol aqueous solution comprising 70 to 95% of a lower alcohol selected from the group consisting of methanol, ethanol and propanol.

Another object of the present invention is to provide a use of a plant extract of the family Cyperaceae for preparing a composition for promoting cell growth, repairing cells, protecting cells, and having anti-oxidation and anti-inflammatory effects, wherein the plant extract of the family Cyperaceae is the plant extract of the family Cyperaceae described above.

The use as described above includes administering the composition to a desired individual to achieve an effective concentration of the extract of the Herba Lysimachiae to promote cell growth, repair cells, protect cells, and provide anti-oxidation and anti-inflammation, wherein the effective concentration is 25 to 500 μg/mL.

Another object of the present invention is to provide an Ophioglossaceae plant extract microcapsule, which includes: an Ophioglossaceae plant extract as described in any one of claims 1 to 6. A plant extract of the family Herbaceae; and an oily microcapsule coating the plant extract of a plant of the family Amaranthaceae, wherein the oily microcapsule has an edible oil selected from the group consisting of olive oil, bitter camellia oil, sesame oil, and flax oil. , a group consisting of canola oil, soybean oil, peanut oil, corn germ oil, salad oil, palm oil, sunflower oil and fish oil.

The above-mentioned microcapsules of extracts of plants of the family Ampelopsis, wherein the oily microcapsules further comprise an antioxidant, which is a fat-soluble antioxidant including vitamin E, vitamin E derivatives, vitamin A or vitamin A derivatives.

The microcapsules of extracts of plants of the family Ampelopsis as described above, wherein the preparation method of the microcapsules of extracts of plants of the family Ampelopsis comprises the following steps: (a) dissolving a water-soluble polymer in a polar solvent to obtain a first solution; (b) uniformly mixing the extract of the plant of the family Ampelopsis with another water-soluble polymer to form a first mixed solution; (c) adding the first mixed solution to the first solution, and stirring uniformly at a first temperature to obtain a second mixed solution; (d) A cross-linking agent is added to the second mixed solution, and the mixture is stirred at a constant temperature to obtain a third mixed solution, which contains aqueous microspheres having the extract of the Herba Lycopodii and the water-soluble polymer; and (e) the third mixed solution is poured into the edible oil, and the mixture is stirred at a constant temperature to form an oily microcapsule to cover the aqueous microspheres, so as to obtain Herba Lycopodii plant extract microcapsules, wherein the cross-linking agent includes saponin, sodium bicarbonate, sodium carbonate, sodium acetate, sodium oxalate or sodium lactate.

In the above-mentioned microcapsules of extracts from plants of the family Psoralea corylifolia, the water-soluble polymer comprises poloxamer 188, polyvinyl pyrrolidone K30, polyvinyl pyrrolidone VA64, HPC-L or HPC-SSL.

Yet another object of the present invention is to provide a use of microcapsules of plant extracts from the family Herbaceae for preparing a composition that can promote cell growth, repair cells, protect cells, antioxidant and anti-inflammation, which includes administering the composition. To reach the effective concentration of the plant extract of the plant of the family Cortaceae to the required individual, in order to promote cell growth, repair cells, protect cells, antioxidant and anti-inflammation, the microcapsule of the plant extract of the plant of the family of the plant is as follows: The effective concentration of the aforementioned plant extract microcapsules is 25 to 500 μg/mL.

One embodiment of the present invention is to provide an Ophioglossaceae plant extract, which has the effects of promoting cell growth, repairing cells, protecting cells, anti-oxidation and anti-inflammatory, and comprises at least 10 to 40 wt% of total flavonoid active molecules and 3 to 10 wt% of total polyphenol active molecules, based on the total weight of the extract, wherein the extract is extracted from an Ophioglossaceae plant; preferably, the Ophioglossaceae plant extract comprises at least 15 to 35 wt% of total flavonoid active molecules and 5 to 8 wt% of total polyphenol active molecules, and more preferably 16 to 33 wt% of total flavonoid active molecules and 5.2 to 7.7 wt% of total polyphenol active molecules, based on the total weight of the extract.

Specifically, in the present invention, the so-called total flavonoid active molecule content is the content per unit weight calculated by testing the bottle grass plant extract using catechin as a standard, and the so-called total polyphenol active molecule is gallic acid. acid) as a standard to detect the content per unit weight of the bottle of Herba Lycopodii plant extract. In some specific examples, each gram of the Herba Lycopodii plant extract includes at least 60 to 400 mg of total flavonoid active molecules and 30 to 100 mg of total polyphenol active molecules; in other preferred examples, each gram of the Herba Lycopodii plant extract includes at least 150 to 300 mg of total flavonoid active molecules and 50 to 70 mg of total polyphenol active molecules; in other more preferred examples, each gram of the Herba Lycopodii plant extract includes at least 160 to 330 mg of total flavonoid active molecules and 52 to 80 mg of total polyphenol active molecules.

In this embodiment, the Ophioglossaceae plants include Helminthostachys zeylanica , Botrychium lunaria , Ophioglossum austroasiaticum , and Ophioglossum austroasiaticum. Ophioglossum pedunculosum , Ophioglossum petiolatum , Ophioglossum reticulatum or Ophioglossum thermale ; preferably, Ophioglossum family ( Ophioglossaceae ) plants include Helminthostachys zeylanica .

Another embodiment of the present invention provides a method for extracting an extract of a plant of the family Erythropaceae. Please refer to Figure 1A. The method includes the following steps: S1) Soaking a bottle of Erythroxylinaceae plant in an aqueous alcohol solution, To obtain a plant immersion liquid; S2) Vibrate the plant immersion liquid with ultrasonic waves at a temperature to obtain a plant crude extract; S3) Filter the plant crude extract to obtain a bottle of Erbacteriaceae plant extract, Wherein, the ultrasonic oscillation is a multi-stage continuous ultrasonic oscillation (S2'), wherein step S1 includes soaking the whole plant or any plant part selected from the rhizomes, stems and leaves of the plant. , spikes and any combination of two or more; the following is a specific example to illustrate the plant parts of the Herbaceae plant, as shown in Figure 1B: Helminthostachys zeylanica , According to the type, each plant part is divided into rhizome, stem, leaf and spike. The rhizome, as shown by the symbol Rz, is a kind of plant with a similar appearance to the root. The abnormal stem is a plant part that grows underground and has a plurality of fleshy thick root structures; the stem, as shown by the symbol St, is an elongated plant part that grows on the ground; the leaves are shown by the symbol Lf , growing on the top of the stem; the spore spike, as indicated by the label Sp, is a vertical elongated structure growing on the top of the stem, with a plurality of sporangia; in various embodiments, the The rhizome of the plant of the family Herbaceae is soaked and extracted.

Specifically, please continue to refer to Figure 1A. The multi-stage continuous ultrasonic oscillation (S2') includes: a first stage, which includes continuously oscillating the plant soaking solution for a first oscillation time T ₁ , and then pausing to oscillate the plant soaking solution. The plant soaking liquid reaches a first pause time t ₁ ; a second stage, which includes continuously shaking the plant soaking liquid for a second shaking time T ₂ , and then pausing to shake the plant soaking liquid for a second pause time t ₂ ; a third stage, which includes continuing to shake the plant soaking liquid for a third shaking time T ₃ , and then pausing to shake the plant soaking liquid for a third pause time t ₃ ; a fourth stage, which includes continuing to shake the plant soaking liquid The liquid reaches a fourth shaking time T ₄ , and then the plant soaking liquid is paused to shake for a fourth pause time t ₄ ; and a fifth stage includes continuous shaking of the plant soaking liquid for a fifth shaking time T ₅ , and then Pause and shake the plant soaking solution for a fifth pause time t ₅ , where T ₁ ＜ T ₂ ＜ T ₃ ＜ T ₄ ＜ T ₅ or T ₁ ＞ T ₂ ＞ T ₃ ＞ T ₄ ＞ T ₅ , and t ₁ ＜t ₂ ＜t ₃ ＜t ₄ ＜t ₅ .

In some embodiments, T ₁ is between 0 and 30 minutes but not 0 minutes, T ₂ is between 0 and 60 minutes but not 0 minutes, T ₃ is between 0 and 70 minutes but not 0 minutes, T ₄ Between 0 and 120 minutes but not 0 minutes, T ₅ between 0 and 180 minutes but not 0 minutes, and t ₁ between 0 and 60 minutes but not 0 minutes, t ₂ between 6 and 60 minutes, t ₃ is between 7 and 70 minutes, t ₄ is between 8 and 180 minutes, t ₅ is between 9 and 120 minutes; preferably, T ₁ is between 0 and 30 minutes but not 0 minutes, and T ₂ is between 9 to 60 minutes, T ₃ between 8 and 70 minutes, T ₄ between 7 and 120 minutes, T ₅ between 6 and 180 minutes, and t ₁ between 0 and 60 minutes but not 0 minutes, t ₂ between 6 to 60 minutes, t ₃ between 7 and 70 minutes, t ₄ between 8 and 180 minutes, t ₅ between 9 and 120 minutes; more preferably, T ₁ between 0 and 30 minutes but not 0 minutes, T ₂ is between 9 and 40 minutes, T ₃ is between 8 and 50 minutes, T ₄ is between 7 and 80 minutes, T ₅ is between 6 and 100 minutes, and t ₁ is between 0 and 60 minutes but not 0 minutes , t ₂ is between 6 and 60 minutes, t ₃ is between 7 and 70 minutes, t ₄ is between 8 and 80 minutes, and t ₅ is between 9 and 120 minutes.

In the present embodiment, the plant of Ophioglossaceae includes Helminthostachys zeylanica, Botrychium lunaria , Helminthostachys zeylanica , Ophioglossum austroasiaticum , Ophioglossum pedunculosum , Ophioglossum petiolatum , Ophioglossum reticulatum or Ophioglossum thermale , preferably Helminthostachys zeylanica .

In these embodiments, the Echinaceae plant can be in a crushed state or in its original state, and is not particularly limited; in the so-called crushed state, the Echinaceae plant is washed and air-dried or dried, and then the required The plant parts are converted into a pulverized state by crushing, grinding or chopping to facilitate subsequent uniform mixing with the solvent.

In these embodiments, the temperature is 20 to 35°C, preferably 22 to 30°C, and more preferably 25°C; the plant soaking liquid includes 1 to 10 parts by weight of a plant of the family Cyperaceae and 15 to 150 parts by weight of an alcohol aqueous solution; in these embodiments, the alcohol aqueous solution includes 70 to 99% of a lower alcohol selected from the group consisting of methanol, ethanol and propanol; the alcohol aqueous solution is preferably 75 to 95% of an ethanol aqueous solution, and more preferably 95% of an ethanol aqueous solution.

In this embodiment, please continue to refer to FIG. 1A , the extraction method further includes the following steps: S4) concentrating the extract of the Herba Lysimachiae to obtain a concentrated extract; and S5) freeze-drying the concentrated extract to obtain a concentrated extract in the form of a dry powder.

Another embodiment of the present invention is to provide a use of a plant extract of the family Cyperaceae for preparing a composition for promoting cell growth, repairing cells, protecting cells, and anti-oxidation and anti-inflammation, wherein the plant extract of the family Cyperaceae is the plant extract of the family Cyperaceae described above or the plant extract of the family Cyperaceae prepared by the extraction method described above.

In various embodiments of this embodiment, the use includes administering the composition to the desired individual to reach an effective concentration of the plant extract of the family Echinacea to promote cell growth, repair cells, protect cells, and resist oxidation. Or anti-inflammatory, wherein the effective concentration is 25 to 500 μg/mL; preferably, the effective concentration is 100 to 400 μg/mL.

Yet another embodiment of the present invention is to provide a microcapsule of a plant extract from a plant of the family Erythropaceae, which includes a plant extract of a plant of the family Erythropaceae and an oily microcapsule, wherein the oily microcapsule coats the plant extract of the family Erythraceae. The grass plant extract is the grass plant extract as mentioned above or the grass plant extract prepared by the method as mentioned above.

In various embodiments, the oily microcapsules have an edible oil selected from the group consisting of olive oil, tea oil, sesame oil, flax oil, canola oil, soybean oil, peanut oil, corn germ oil, salad oil, palm oil, sunflower oil, and A group consisting of fish oil.

In various embodiments, the oily microcapsules further comprise an antioxidant, which is a fat-soluble antioxidant including vitamin E, vitamin E derivatives, vitamin A or vitamin A derivatives.

Specifically, please refer to FIG. 1B , the preparation method of the microcapsule of the extract of the phytoplankton family includes the following steps: (a) dissolving a water-soluble polymer in a polar solvent to obtain a first solution; (b) uniformly mixing the extract of the phytoplankton family with another water-soluble polymer to form a first mixed solution; (c) adding the first mixed solution to the first solution, and stirring uniformly at a first temperature to obtain a second mixed solution; (d) adding a crosslinking agent to the second mixed solution, and stirring at a constant temperature to obtain a third mixed solution, which contains aqueous microspheres having the extract of the phytoplankton family and the water-soluble polymer; and (e) Pour the third mixed solution into the edible oil and fat, and stir at a constant temperature to form an oily microcapsule to cover the aqueous microsphere, so as to obtain a bottle of microcapsules of the Herba Herbaceae extract.

In various embodiments, the crosslinking agent comprises saponin, sodium bicarbonate, sodium carbonate, sodium acetate, sodium oxalate or sodium lactate; preferably, the crosslinking agent is sodium bicarbonate.

In various embodiments, the water-soluble polymer includes poloxamer 188, polyvinyl pyrrolidone K30, polyvinyl pyrrolidone VA64, HPC-L or HPC-SSL.

In some embodiments, the fat is dissolved in an antioxidant system including vitamin E, vitamin E derivatives, vitamin A or vitamin A derivatives.

In some embodiments, the first temperature is 60 to 85°C, wherein steps (c), (d), and (e) are stirred for 10 to 60 minutes, respectively. Preferably, the first temperature is 70°C, and steps (c), (d), and (e) are stirred for 20 to 45 minutes, respectively.

In these embodiments, in the step (b), the volume ratio of the first solution to the extract of the Herba Lycopodii family is (1 to 10):1; in the step (c), the volume ratio of the first mixed solution to another first solution is 1:(1 to 10); in the step (e), the volume ratio of the third mixed solution to the oil is (1 to 10):1.

Yet another embodiment of the present invention is the use of a microcapsule of a plant extract of the Herba family for preparing a composition that promotes cell growth, repairs cells, protects cells, is antioxidant and anti-inflammatory, which includes administering the composition The effective concentration of the plant extract of the plant of the family Erythroxylum aeruginosa is reached to the required individual to promote cell growth, repair cells, protect cells, antioxidant or anti-inflammation, wherein the microcapsule of the plant extract of the plant Erythroxylinum nuciferae is as described above. The effective concentration of the microcapsules of the plant extract of Boeraceae is 25 to 500 μg/mL; preferably, the effective concentration is 100 to 400 μg/mL.

According to the present invention, the compositions may further comprise an acceptable adjuvant for the preparation of skin care products, and the acceptable adjuvant may include one or more agents selected from the following: solvents, gelling agents, active agents, preservatives, antioxidants, screening agents, chelating agents, surfactants, dyeing agents, thickeners, fillers, fragrances and odor absorbents.

According to the present invention, the skin care products can be manufactured into a form suitable for skin care or makeup using techniques well known to those skilled in the art, including but not limited to aqueous solutions, alcohol-water solutions or oily solutions, or water-in-oil, oil-in-water or composite emulsions, gels, ointments, creams, masks, patches, plasters, wipes, powders, aerosols, sprays, lotions, emulsions, pastes, foams, dispersions, drops, mousse, sunscreen, lotions, foundations, makeup removers and soaps, etc.

According to the present invention, the skin care product can also be used in combination with one or more external agents with known activity, wherein the external agents can include whitening agents such as vitamin A acid, catechin, kojic acid, and arbutin. or vitamin C, humectants, anti-inflammatory agents, bactericides, ultraviolet absorbers, plant extracts such as aloe vera extracts, skin nutrients, anesthetics, anti-acne agents, anti-itch agents, analgesics, anti-dermatitis agents, anti-inflammatory agents Keratinizing agents, anti-dry skin agents, antiperspirants, anti-aging agents, anti-wrinkle agents, anti-seborrhea agents, wound treatment agents, corticosteroids and hormones.

According to the present invention, these compositions can also be used as food additives, added to food raw materials in a known manner, or added during the processing of food raw materials, and formulated with any edible material into food products for human and non-human animals to consume; specifically, the food products include but are not limited to beverages, fermented foods, baked foods, health foods or dietary supplements.

In addition, the following several examples are used to further prove the scope of practical application of the present invention, but are not intended to limit the scope of protection of the present invention. All modifications and improvements based on the spirit of the present invention are within the scope of the present invention. Declaration of scope of protection. Table 1 Multi-stage ultrasonic vibration Example Original/Crushed Solvent Liquid to solid ratio (weight ratio) temperature first stage second stage The third stage Stage 4 The fifth stage 1 Original state 95% ethanol 15~150: 1~10 room temperature Vibration time (minutes) 0＜T ₁ ≦30 0＜T ₂ ≦40 0＜T ₃ ≦50 0＜T ₄ ≦80 0＜T ₅ ≦100 Pause time (minutes) 0＜ _t1 ≦60 10＜ _t2 ≦60 20＜ _t3 ≦70 30＜ _t4 ≦80 60＜t ₅ ≦120 2 smash 95% ethanol 15~150: 1~10 room temperature Vibration time (minutes) 0＜T ₁ ≦50 0＜T ₂ ≦60 0＜T ₃ ≦70 0＜T ₄ ≦120 0＜T ₅ ≦180 Pause time (minutes) 5＜t ₁ ≦30 10＜ _t2 ≦30 30＜ _t3 ≦60 60＜ _t4 ≦180 30＜ _t5 ≦120 3 smash 75% ethanol 15~150: 1~10 room temperature Vibration time (minutes) 0＜T ₁ ≦20 0＜T ₂ ≦19 0＜T ₃ ≦18 0＜T ₄ ≦17 0＜T ₅ ≦16 Pause time (minutes) 5＜t ₁ ≦10 6＜ _t2 ≦11 7＜t ₃ ≦17 8＜t ₄ ≦18 9＜t ₅ ≦19 4 Original state 75% ethanol 15~150: 1~10 room temperature Vibration time (minutes) 0＜T ₁ ≦20 0＜T ₂ ≦19 0＜T ₃ ≦18 0＜T ₄ ≦17 0＜T ₅ ≦16 Pause time (minutes) 5＜t ₁ ≦10 6＜ _t2 ≦11 7＜t ₃ ≦17 8＜t ₄ ≦18 9＜t ₅ ≦19 5 Original state 55% ethanol 15~150: 1~10 room temperature Vibration time (minutes) 0＜T ₁ ≦20 0＜T ₂ ≦19 0＜T ₃ ≦18 0＜T ₄ ≦17 0＜T ₅ ≦16 Pause time (minutes) 5＜t ₁ ≦10 6＜ _t2 ≦11 7＜t ₃ ≦17 8＜t ₄ ≦18 9＜t ₅ ≦19 Comparative example Original state pure water - 40 to 50 Soaking time (hour) 1 to 8 hours

Ceylon Seven Fingers Extract

Please refer to Table 1, which details the specific parameters of Examples 1 to 5; generally speaking, the experimental method is to mix the rhizome of Pterostilbene with a solvent to obtain a mixture of plant parts, and Multi-stage ultrasonic extraction is carried out at room temperature, which is divided into 5 stages, namely the first stage, the second stage, the third stage, the fourth stage and the fifth stage. In each stage, the plants are shaken in sequence. After the mixed liquid reaches the set shaking time, the shaking is paused to reach the set stopping time; after completing the multi-stage ultrasonic extraction, filter the extraction liquid obtained after shaking to obtain the filtrate; then concentrate the filtrate, and then concentrate it. The extract is finally freeze-dried to obtain dry powdery extract of Ceylon Fern.

Please refer to the column below Table 1. In order to compare the active substance extraction rate and biological function of the non-multi-stage ultrasonic extraction of the Ceylon seven-fingered fern extract in subsequent experiments, the rhizome of the original Ceylon seven-fingered fern was soaked in pure water as a comparison example. The extract obtained after soaking was also filtered, concentrated, and freeze-dried for use in subsequent experiments.

Experimental example 1 : Evaluation of the effect of extracts on fibroblast proliferation efficiency

In a 96-well plate, 10,000 human dermal fibroblasts (Human Dermal Fibroblasts) were planted in each culture well, and cultured overnight with 100µl cell culture medium (10% FBS MEM medium) to allow cells to adhere; remove the culture medium from each culture well After the solution was washed twice with PBS, 100 µl of cell culture solution containing extracts of different concentrations was added again, and the 96-well plate was placed in a cell culture incubator for reaction for 24 or 48 hours. Cell proliferation was detected using Sulforhodamine B colorimetric assay (Sulforhodamine B, SRB assay).

In Experimental Example 1, the SRB Kit (Cat. ab235935) provided by Abcam was used. The SRB colorimetric method is briefly described as follows: 50 μl of TCA solution (30%, w/v) pre-cooled to 4°C was added to each culture well to fix the cells. , the final concentration of the TCA solution is 10%; let it stand for 5 minutes, then move it to a 4°C refrigerator to fix for 1 hour, rinse it with deionized water 5 times, and dry it at room temperature; after drying, add SRB to each culture well 45 μl of dye solution. After staining for 30 minutes, pour out the dye solution, rinse the culture wells 4 times with Wash buffer to remove unbound dye, and dry at room temperature; finally, add 200 μL of solubilization buffer (Solubilization buffer) to wash and place Shake on a horizontal oscillator for 4 minutes and repeat four times. The light absorption value of the washed cells is measured at 565 nm. Table 2 Treatment concentration (%) Cell proliferation rate (%) 0.266 0.301 Example 1 288.6 500.0 Example 2 123.7 247.6 Example 3 129.4 293.6

The results of the cell proliferation experiment are shown in Table 2 and Figure 2. The extract prepared in Example 1 has excellent cell proliferation effect. At a concentration of 0.266%, it can cause cell proliferation by 288.6%. At a concentration of 0.301%, It can cause cell proliferation by 500%, which is equivalent to having 5 times the cell proliferation effect; the extract prepared in Example 2 can cause cell proliferation by 123.76% at a concentration of 0.266%, and can cause cell proliferation by 123.76% at a concentration of 0.301%. 247.58%, equivalent to 2.5 times the cell proliferation effect, and also has a good cell proliferation effect; the extract prepared in Example 3 can cause cell proliferation by 129.4% at a concentration of 0.266%, and at a concentration of 0.301%, It can cause cell proliferation by 293.6%, which is equivalent to nearly 3 times the cell proliferation effect, and its cell proliferation effect is also quite excellent. Although Comparative Example 1 extracted with pure water also has the effect of cell proliferation, it only makes cells more effective at a concentration of 0.266%. The cell proliferation rate is 119.5%. At a concentration of 0.301%, the cell proliferation rate is only 142%, which is not as good as the cell proliferation-promoting effect obtained by alcohol extraction in Examples 1 to 3.

Experimental example 2 : wound healing test (Wound healing assay)

Culture 5 × 10 ⁴ human dermal fibroblasts (HDF) cells in a 24-well culture plate. Add 500 μl MEM culture medium to each culture well and culture for 24 hours. When the culture time is up, use the tip of a 1 ml micropipette. (tip) Draw a small groove in the center of the cells at the bottom of the circular plate, as shown in Figure 3 for 0 hours; then, draw out the culture medium from the culture well, add PBS to wash twice; then add 500µl of different Cell culture medium with concentrations of extracts (0.226%, 0.113%); and then continued to culture for 24 hours and 48 hours. The effects of different concentrations of extracts on the degree of cell healing were observed under a microscope at these time points.

Please continue to refer to Figure 3. After 48 hours of culture, compared with the untreated control group, the cell healing efficiency of the extract prepared in Example 1 was improved, and as the treatment concentration increased, the cell healing efficiency increased. The higher the healing efficiency; the control group that was not treated with any extract had a wound healing rate of only 12.8% after 48 hours, while the group treated with a concentration of 0.113% achieved a wound healing rate of 29.2% after 48 hours, and the treatment concentration was 0.226 % group even reached a wound healing rate of 78.6%.

Experimental example 3 ：Antioxidant Assessment - Clear DPPH The Effects of Free Radicals

In this experiment, the scavenging effect of Examples 1, 4 and 5 on DPPH free radicals was tested to evaluate the effectiveness of the samples in scavenging free radicals. The specific method was to add 50 μL of the sample of different concentrations to each well of a 96-well plate, and then add 150 μL of a 200 μM DPPH solution for reaction, and place it at 25°C for 15 minutes. The change in absorbance at a wavelength of 515 nm within the aforementioned 15 minutes was measured using a microplate reader (Molecular Devices Spectra Max M2 Microplate Reader). The absorbance values of Examples 1, 4, 5 and Comparative Example 1 after the reaction were different, and each was compared with the control group to convert the relative ability of each Example to scavenging DPPH. Table 3 DPPH removal efficiency (%) Treatment concentration (μg/ml) Embodiment 1 Control group 5 25.0±2.8 53.7±1.3 10 35.8±1.4 60.1±1.3 15 52.6±2.8 65.2±1.2 20 70.2±3.4 91.0±0.7 25 81.5±3.3 95.1±0.9 50 99.2±1.1 96.9±1.7

In Experimental Example 3, vitamin C (Vit C) is used as the control group; please refer to Table 3 and Figure 4A. The experimental results show that in the low concentration range (5 to 25 μg/ml), Example 1 is effective in scavenging DPPH free radicals. The efficiency is lower than Vit C, but when the concentration reaches 50 μg/ml, Vit C can scavenge 96.9% of DPPH free radicals, but the efficiency of scavenging DPPH free radicals in Example 1 can reach 99.2%, which shows a higher ability to scavenge DPPH free radicals. effect. Table 4 DPPH removal efficiency (%) Treatment concentration (μg/ml) control group Example 1 Example 4 Example 5 Comparative example 1 25 95.1±0.9 81.5±3.3 15.8±2.8 7.9±0.3 0.1±0.1 50 96.9±1.7 99.2±1.1 62.8±2.7 24.5±1.4 1.9±1.0 100 98.7±2.2 99.4±0.1 90.5±1.0 49.0±1.6 5.0±1.0 200 99.1±1.5 99.7±0.3 92.2±1.0 89.2±1.2 22.9±3.3 400 99.4±1.4 99.9±0.9 95.5±1.0 94.1±3.4 63.8±0.4

Please refer to Table 4 and Figure 4B. Vit C was used as the control group to compare the scavenging efficiency of Examples 1, 4, 5 and Comparative Example 1 for DPPH free radicals. Among them, when the concentration of Examples 4, 5 and Comparative Example was lower than 50 μg/ml, The scavenging efficiency of DPPH free radicals was not obvious, being lower than 1.9±1.0%, 24.5±1.4%, and 62.8±2.7%, respectively. In the concentration range of 100 to 400 μg/ml, Example 1 still exhibited the highest scavenging efficiency of DPPH free radicals, reaching 99.9±0.9% at a concentration of 400 μg/ml, while Examples 4 and 5 reached 95.5±1.0% and 94.1±3.4%, respectively. In contrast, Comparative Example 1 extracted with pure water only reached 63.8±0.4%, and the effect was not significant.

Experimental example 4 : Antioxidant assessment - Clear ABTS The Effects of Free Radicals

In this experimental example, the scavenging effect of Examples 1, 4 and 5 on ABTS free radicals was tested to evaluate the effectiveness of the samples in scavenging free radicals; the specific method was to prepare 14 mM ABTS solution and 4.9 mM K ₂ S with deionized water respectively. ₂ O ₈ solution; take equal amounts of ABTS solution and K ₂ S ₂ O ₈ solution, mix evenly at 25°C in a dark environment for 12 to 24 hours and set aside; when performing experiments, take this mixed solution and dilute it with deionized water 25 times to form the ABTS analysis solution, measure the absorbance value at 734 nm to be about 0.7, store it away from light and use it on the day of the experiment; then, add 100 μl of the sample to be tested including Vit to each microwell of the 96-well plate. C. Example 1, Example 4, Example 5 and Comparative Example 1, then add 100 μl ABTS analysis solution and react at 25°C for 20 minutes; measure with a microplate reader (Molecular Devices Spectra Max M2 Microplate Reader) Measure the changes in the absorbance value at a wavelength of 734 nm after the aforementioned reaction for 20 minutes; the absorbance values after the reaction of each embodiment 1, 4, 5 and comparative example 1 are different, and each is compared with the control group to convert the ABTS removal value of each embodiment Relative ability. table 5 ABTS clearance efficiency (%) concentration (μg/ml) Example 1 control group 5 35.9±1.1 43.7±1.8 10 60.1±2.3 64.8±2.1 15 73.8±1.5 80.9±1.3 20 83.6±1.1 88.1±2.0 25 98.1±1.3 93.7±1.4 50 99.1±1.2 96.6±2.3

In Experimental Example 4, vitamin C (Vit C) was used as a control group; see Table 5 and FIG. 5A . The experimental results show that in the low concentration range (5 to 20 μg/ml), the efficiency of Example 1 in scavenging ABTS free radicals is lower than that of Vit C. However, when the concentration reaches 25 to 50 μg/ml, Vit C can scaveng 93.7±1.4% and 96.6±2.3% of ABTS free radicals, respectively. However, the efficiency of Example 1 in scavenging ABTS free radicals can reach 98.1±1.3% and 99.1±1.2%, respectively, which shows a higher efficacy in scavenging ABTS free radicals.

Please refer to Table 6 and Figure 5B. Vit C is also used as the control group to compare the scavenging efficiency of ABTS free radicals in Examples 1, 4, 5 and Comparative Example 1. Among them, the concentrations of Examples 4, 5 and Comparative Example 1 are When the interval is lower than 50 μg/ml, the scavenging efficiency of ABTS free radicals is not obvious, which are lower than 54.0±1.5%, 60.2±1.7%, and 26.7±1.2 respectively; in the concentration interval between 100 and 400 μg/ml, Example 1 It still shows the highest DPPH free radical scavenging efficiency, achieving the complete ABTS scavenging effect (100.8±1.2%) at a concentration of 100 μg/ml, while Examples 4 and 5 reached 100.0±1.8% and 100.0±1.8% respectively at a concentration of 400 μg/ml. The ABTS removal efficiency is 99.4±1.5%; on the other hand, Comparative Example 1 extracted with pure water only reaches 43.5±2.6% at a concentration of 100 μg/ml, and the effect is not significant. However, it can reach 97.8±0.5% at a high concentration of 400 μg/ml. The ABTS removal efficiency is still not as good as the ABTS removal efficiency of Examples 1, 4 and 5. Table 6 ABTS clearance efficiency (%) concentration (μg/ml) control group Example 1 Example 4 Example 5 Comparative example 1 25 93.7±1.4 98.1±1.4 42.3±1.9 39.6±1.6 21.1±1.5 50 96.6±2.3 99.1±1.3 60.2±1.7 54.0±1.5 26.7±1.2 100 100.9±1.6 100.8±1.2 87.8±2.3 83.1±2.1 43.5±2.6 200 100.3±2.3 100.8±1.1 98.8±1.5 98.6±0.7 72.7±1.5 400 100.2±2.5 100.9±1.6 100.0±1.8 99.4±1.5 97.8±0.5

Experimental example 5 ：Total flavonoids and total polyphenols content detection

The extract powders of Examples 1 and 4 were prepared and mixed with 1 mg/mL of the test sample and 10 μL of 0.05M NaNO ₂ evenly. The reaction was carried out in the dark at room temperature for 6 minutes, and then 10 μL of 1M Al(NO ₃ ) ₃ was injected. And react at room temperature for 5 minutes in the dark. Finally, inject 80 μL of 1M NaOH, and react at room temperature for 30 minutes in the dark. Record the absorbance value of the mixture at a wavelength of 510 nm, and use Catechin as the standard for calculation. Flavonoid content in Examples 1 and 4.

On the other hand, take the extract powders of Examples 1 and 4 and prepare a test sample of 1 mg/mL respectively, and sequentially inject 20 μL of sample and 90 μL of 10% Folin-Ciocalteu into a 96-well plate at room temperature. React at room temperature for 5 minutes in the dark, and then inject 90 μL of 5% Na ₂ CO _3. After the same reaction at room temperature in the dark for 2 hours, record the absorbance value of the sample to be tested at a wavelength of 750 nm, and add it with gallic acid (Gallic acid). acid) was used as a standard substance to calculate the total phenolic content of Examples 1 and 4.

Please refer to Table 7, which respectively shows the calculated total flavonoid contents and total polyphenol contents of Examples 1, 4 and Comparative Example 1; wherein, the total flavonoid contents are 328.8±1.3 mg/g, 160.8±1.8 mg/g and 68.3±1.8 mg/g, respectively, and the total polyphenol contents are 76.3±2.0 mg/g, 52.3±1.6 mg/g and 36.1±1.5, respectively; obviously, compared with Comparative Example 1 which is extracted with pure water, the alcohol extraction can obtain higher total flavonoid and total polyphenol active substances. Table 7 Total flavonoid content (mg/g) Total polyphenol content (mg/g) Embodiment 1 328.8±1.3 76.3±2.0 Embodiment 4 160.8±1.8 52.3±1.6 Comparison Example 1 68.3±1.8 36.1±1.5

Experimental example 6 : Testing of the UV protection efficacy of Ceylon sphenopsis extract

In Experimental Example 6, the efficacy of the Ceylon Fern extract in protecting cells from UVB damage was evaluated; the experimental method specifically included culturing fibroblast CCD966SK (1×10 ⁴ /well) in a 96-well plate, and in Incubate in a 37°C and 5% _CO2 incubator for 24 hours.

In the non-irradiated group (without UVB irradiation), the cell supernatant was removed and replaced with fresh culture medium, followed by 4 hours of culture, followed by washing with PBS and placing in fresh culture medium for another 4 hours before survival analysis.

In the control group (without the addition of the extract of Pteris crenata), the cell supernatant was removed and replaced with fresh culture medium, and then the cells were cultured for 4 hours. The supernatant was removed and washed with PBS. Then the PBS was removed and UVB irradiation was started. The UVB energy used was 20 mJ/cm ² . Fresh culture medium was added after UVB irradiation. The cells were cultured for 4 hours and then the cell viability was analyzed.

In the treatment group (with the addition of Pteris sinensis extract), the cell supernatant was removed, Example 1 and Comparative Example 1 were respectively added to the fresh culture medium, and the concentrations of Example 1 or Comparative Example 1 were respectively prepared to reach 100, 300 and 500 μg/ml culture medium, and culture the cells with these culture medium for 4 hours, remove the supernatant and wash with PBS, then remove the PBS, conduct UVB irradiation (20 mJ/cm ² ) respectively, and then Then add fresh culture medium containing Example 1 or Comparative Example 1 at concentrations of 100, 300, and 500 μg/ml, continue culturing for 4 hours, and then perform cell viability analysis.

Cell viability analysis includes removing the old culture medium, washing once with PBS, replacing with new culture medium, adding culture medium containing 0.5 mg/ml MTT (3-(4,5-cimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide), and reacting in a cell culture incubator for 3 hours; after removing the MTT solution, 200µl DMSO was added to dissolve the formazan precipitate, and then the absorbance value at a wavelength of 550 nm was analyzed by ELISA to calculate its viability. Table 8 Treatment concentration (μg/mL) Cell viability (%) Unirradiated group Control group Processing Group 0 0 100 300 500 Embodiment 1 100.0±1.2 67.1±1.8 78.5±2.0 85.9±1.9 98.9±1.6 Comparison Example 1 100.0±0.9 67.1±1.8 68.3±2.0 69.3±1.3 67.9±2.3

Please refer to Table 8. The viability of the unirradiated tissue cells is 100%. In an environment without adding the extract for protection, after UVB light irradiation, the cell viability decreased to 67.1±1.8%. The cell protection experiment was carried out by adding different concentrations of Example 1 to the cells. The results showed that the extract of Seven Fingers Pteris Ceylon was helpful in protecting cells from the damage caused by UVB irradiation. Under the protection treatment of treatment concentrations of 100μg/mL, 300μg/mL and 500μg/mL, the cell viability after UVB irradiation reached 78.5±2.0%, 85.9±1.9% and 98.9±1.6%, respectively. The results showed that the extract of Ceylonema fern prepared according to Example 1 has the function of protecting cells from damage caused by UVB ultraviolet light. On the other hand, the cell survival rates of the Ceylonema fern extract obtained in Comparative Example 1 after UVB irradiation were only 68.3±2.0%, 69.3±1.3% and 67.9±2.3, respectively, under the protection treatments of treatment concentrations of 100μg/mL, 300μg/mL and 500μg/mL, which were not significant compared with the control group. It is obvious that the extract of Ceylonema fern extracted with pure water cannot effectively protect cells from damage caused by UVB ultraviolet light.

Experimental example 7 : Anti-inflammatory efficacy test

In Experiment 7, LPS was used to induce the expression of nitric oxide in mouse RAW264.7 cells to evaluate the anti-inflammatory effect of the extract of Pteris cretica. The experimental method specifically includes planting RAW264.7 cells at a number of 5×10 ⁴ cells/well in a 96-well culture plate, culturing them overnight with 100 μl of DMEM culture medium containing 10% FBS, removing the culture medium and adding culture medium containing 50 ng/ml LPS and different concentrations of Pteris cretica extract; after reacting for 24 hours, taking out 100 μl of the culture medium and adding 100 μl of Griess reagent (concentration of 0.04 g/ml) to test the content of nitrite, a metabolite of NO, in the culture medium after LPS-induced mouse RAW264.7 cells.

Please refer to Table 9 and Figure 6. In the treatment concentration range of 50 to 200 μg/mL, Example 1 showed stronger anti-inflammatory effects than Comparative Example 1. Example 1 inhibited NO generation rate by 68.5±1.2% at a treatment concentration of 200 μg/ml, while Comparative Example 1 only reached 32.7±1.8%. The control group was treated with rutin at a concentration of 200 μg/mL, and its NO generation rate was only inhibited by 56.8%. This shows that the alcohol-extracted extract of the seven-fingered fern not only showed a higher NO generation rate inhibition effect compared to the pure water extraction, but also showed a better NO generation rate inhibition effect than the positive control group rutin, indicating that it has a good anti-inflammatory effect on cells. Table 9 NO generation inhibition rate (%) Treatment concentration (μg/ml) Control group Embodiment 1 Comparison Example 1 50 29.6±1.1 36.3±0.9 25.6±1.3 100 39.8±0.9 53.6±1.5 28.9±1.6 200 56.8±1.3 68.5±1.2 32.7±1.8

The plant extract of the family Pteris cerana provided by the present invention has active ingredients of Pteris cerana including flavonoids and total polyphenols, and exhibits the effects of promoting cell proliferation, repair, anti-oxidation, anti-ultraviolet damage and anti-inflammation in the low concentration range (25 to 50 μg/mL). In addition, in the high concentration range (100 to 500 μg/mL), it not only has the above-mentioned effects, but also has no cytotoxicity, making it safe in application aspects such as cosmetics, skin care products or food.

Since the extract of the plant of the family Pinellaceae provided by the present invention can exert the aforementioned effects at only a low concentration, more dosage selectivity can be provided in subsequent applications, so that the amount added can be reduced during the processing of cosmetics, skin care products or foods, so that the final product can have more room for other added ingredients. The invention has wide applicability and considerable industrial value.

S1 to S5: steps

S2’: Step

a to e: Steps

Figure 1A is a flowchart illustrating the extraction method of the plant extract of the family Erythropaceae provided by the present invention; Figure 1B shows the specific plant parts of the plant Erythraceae provided by the present invention; Figure 1C is an illustration of the plant extract of Erythraceae provided by the present invention. Flowchart of the preparation method of plant extract microcapsules; Figure 2 is a bar chart to illustrate the effect of plant extracts of the family Erythroxylinaceae on cell survival rate under different extraction conditions; Figure 3 is a microscope image to illustrate different treatments The effect of plant extracts from Erythroxylinaceae on cell wound healing at different concentrations; Figures 4A to 4B are bar graphs illustrating the DPPH removal efficiency of Erythromycinaceae plant extracts at different treatment concentrations; Figures 5A to 5B The bar graph is used to illustrate the ABTS scavenging efficiency of the plant extracts of the family Echinaceae under different treatment concentrations; Figure 6 is the bar chart used to illustrate the inhibitory effect of the plant extracts of the family Echinaceae on NO production under different treatment concentrations.

Claims (8)

A method for extracting plant extracts from a bottle of Herbaceae, including: S1) Soaking the original whole plant or rhizome of a bottle of Herbaceae plant in an alcoholic aqueous solution to obtain a plant soaking liquid, wherein, The Helminthostachys zeylanica plant includes Helminthostachys zeylanica ; S2) The plant soaking liquid is vibrated with ultrasonic waves at a temperature to obtain a crude plant extract, wherein the ultrasonic wave vibration is a polysaccharide Stage continuous ultrasonic oscillation, which includes: a first stage, which includes continuous shaking of the plant soaking liquid for a first shaking time T ₁ , and then pausing to shake the plant soaking liquid for a first pause time t ₁ ; a first stage. The second stage includes continuous shaking of the plant soaking liquid for a second shaking time T ₂ , and then pausing to shake the plant soaking liquid for a second pause time t ₂ ; a third stage, which includes continuous shaking of the plant soaking liquid for a second pause time T 2 a third shaking time T ₃ , then pausing shaking the plant soaking liquid for a third pause time t ₃ ; a fourth stage including continuing to shake the plant soaking liquid for a fourth shaking time T ₄ , and then pausing shaking the plant soaking liquid The plant soaking liquid reaches a fourth pause time t ₄ ; and a fifth stage includes continuously shaking the plant soaking liquid for a fifth shaking time T ₅ , and then pausing to shake the plant soaking liquid for a fifth pause time t ₅ , wherein, T ₁ <T ₂ <T ₃ <T ₄ <T ₅ and t ₁ <t ₂ <t ₃ <t ₄ <t ₅ ; and S3) filter the plant crude extract to obtain a bottle of Erxiaoxiangke Plant extracts. The method as described in claim 1, wherein the Botrychium lunaria plants include Botrychium lunaria, Ophioglossum austroasiaticum , Ophioglossum pedunculosum , Ophioglossum petiolatum , Ophioglossum reticulatum or Ophioglossum thermale . The method as described in claim 1, further comprising the following steps: S4) concentrating the Erythropus plant extract to obtain a concentrated extract; and S5) freeze-drying the concentrated extract, To obtain a concentrated extract in dry powder form. The method as described in claim 1, wherein the temperature is 20 to 35°C, the plant soaking liquid comprises 1 to 10 parts by weight of a plant of the family Aglaonema and 15 to 150 parts by weight of an alcohol aqueous solution, the alcohol aqueous solution comprising 70 to 95% of a lower alcohol selected from the group consisting of methanol, ethanol and propanol. A method for preparing a composition of a plant extract of the family Trichoderma spp. to promote cell growth, repair cells, protect cells, antioxidant and anti-inflammation, which includes administering the composition to a desired individual to reach an effective concentration. An extract of a plant of the family Herbaceae, wherein the extract of a plant of the family Herbaceae is prepared according to the extraction method described in any one of claims 1 to 4. The use as described in claim 5, wherein the effective concentration is 25 to 500 μg/mL. A use of an Ophioglossaceae plant extract for preparing an Ophioglossaceae plant extract microcapsule that promotes cell growth, repairs cells, protects cells, and has anti-oxidation and anti-inflammatory effects, comprising administering the microcapsule to a desired individual to achieve an effective concentration of the Ophioglossaceae plant extract, wherein the Ophioglossaceae plant extract microcapsule comprises: an Ophioglossaceae plant extract prepared by the extraction method described in any one of claims 1 to 4; and an oily microcapsule that encapsulates the Ophioglossaceae plant extract, wherein the oily microcapsule has an edible oil. The use as described in claim 7, wherein the effective concentration is 25 to 500 μg/mL.