阅读说明：本技术 一种密蒙花醇提取物的应用 (Application of alcohol extract of buddleja officinalis ) 是由 姚期凤 姜丹 吴越 王越 蒋耀权 章漳 于 2018-05-29 设计创作，主要内容包括：本发明公开了一种密蒙花醇提取物的应用,具体为在制备抗皮肤衰老基因的表达上调剂中的应用。密蒙花醇提取物可以促进胞外基质合成和稳定基底膜结构完整,稳固皮肤结构的作用,诱导细胞修复相关的及抗衰老相关的基因表达,提高皮肤的结构的稳定和延缓衰老的功效；在特定的浓度范围下,可使皮肤抗衰老相关基因的表达量上调至1.36倍以上,最高可达2.13倍。该发现对于日后研发抗衰老相关的护肤品或者化妆品有重要意义。(the invention discloses application of a buddleja officinalis alcohol extract, in particular to application of the buddleja officinalis alcohol extract in preparation of an expression up-regulating agent of a skin aging resistant gene. The alcohol extract of flos Buddlejae can promote synthesis of extracellular matrix, stabilize basement membrane structure, stabilize skin structure, induce cell repair-related and anti-aging-related gene expression, and improve skin structure stability and antiaging effect; under a specific concentration range, the expression level of the skin anti-aging related gene can be up-regulated to more than 1.36 times, and can reach 2.13 times at most. The discovery is of great significance for developing skin care products or cosmetics related to anti-aging in the future.)

1. an application of alcohol extract of flos Buddlejae (flos Buddleja officinalis Maxim) in preparing expression up-regulator of anti-skin aging gene is provided.

2. The use of claim 1, wherein the anti-skin aging gene is selected from one or more of the SIRT1 gene, FGF7 gene, HBEGF gene, NID1 gene, FBN1 gene and FN1 gene.

3. the use of claim 1, wherein the alcohol extract of buddleja officinalis is an alcohol extract of buddleja officinalis from the group consisting of C1 to C5; preferably, the buddleja officinalis is an ethanol extract, and the ethanol is preferably 70-95% ethanol;

Preferably, the alcohol extract of buddleja officinalis is prepared by the following method comprising:

(1) Pulverizing flos Buddlejae to obtain powder; the flos Buddlejae is preferably flos Buddlejae whole plant;

(2) adding ethanol for extraction, and filtering to obtain filtrate; the extraction temperature is preferably room temperature, and the room temperature is 20-30 ℃;

(3) Evaporating the filtrate to obtain; the degree of evaporation is to dryness.

4. The use according to claim 3, wherein the powder in step (1) has a mesh size of 20 to 60 mesh.

5. The use of claim 3, wherein said ethanol in step (2) is 70% ethanol, and said ethanol is added in an amount of 8-12 times, preferably 10 times, the volume of said pulverized butterflybush flower;

And/or, the extraction times in the step (2) are more than 1 time, preferably 2 times.

6. The use of claim 3, wherein the extraction in step (2) is ultrasonic extraction, and the frequency of the ultrasonic extraction is preferably 400-550W, more preferably 500W;

And/or the ultrasonic treatment time is 25-40min, preferably 30 min.

7. The use according to claim 3, wherein the filtration method in step (2) is centrifugation, the centrifugation speed is preferably 6000-10000rpm, more preferably 8000rpm, and the centrifugation time is preferably 8-12 minutes, more preferably 10 minutes.

8. the use of claim 3, wherein said alcohol extract of Buddleja officinalis is prepared by a method comprising:

(1) Pulverizing dried whole plant of flos Buddlejae to obtain powder with mesh number of 20;

(2) adding 70% ethanol with a volume 10 times of the powder, performing 500W ultrasonic extraction at room temperature for 2 times, each for 30min, centrifuging, filtering, and mixing filtrates; the speed of the centrifugation is 8000rpm, and the time of the centrifugation is 10 minutes;

(3) And (5) rotatably evaporating the filtrate to dryness to obtain the product.

9. The use of claim 8, wherein the alcohol extract of buddleja officinalis is present in the up-regulator of expression of the anti-skin aging gene at a concentration of 0.8 to 12 μ g/ml; preferably 1 to 10. mu.g/ml; wherein, the concentration in the expression up-regulator for HBEGF gene is preferably 0.8-1.2 μ g/ml, more preferably 1 μ g/ml; the concentration in the up-regulator for expression of FGF7 and/or FN1 gene is preferably 8 to 12. mu.g/ml, more preferably 10. mu.g/ml.

10. The use according to any one of claims 1 to 9, wherein the up-regulator of the expression of the anti-skin-aging gene is used in cosmetics or skin care products.

Technical Field

The invention relates to the field of plant extracts and daily chemicals, in particular to application of alcohol extract of buddleja officinalis.

Background

the skin covers the surface of a human body and is in direct contact with the external environment, so that the human body can be protected from being damaged by various external factors, and the human body can be used as a mirror to reflect the health state of the human body. The skin is composed of epidermis, dermis, and subcutaneous tissue, and contains skin appendages, abundant nerves, blood vessels, lymphatic vessels, and muscles. The connective extracellular matrix region of the dermis and epidermis, also known as the Basement Membrane (BM), not only supports the epidermis, but also serves multiple functions, such as the junctions of the dermis and epidermis acting as a selective cellular barrier that generally prevents connective tissue cells from contacting the epidermal cells of the Basement membrane, but allows passage of lymphocytes, macrophages, and neuronal synapses; in addition, many vital phenomena such as cell morphology, cell polarity, cell metabolism, protein distribution in plasma membranes, cell survival, cell proliferation and differentiation, and cell migration are very closely related to the basement membrane. Nestin is an important component of basement membrane and is essential for maintaining the structure and function of basement membrane, which is essential for maintaining normal renewal of skin. The dermis is composed of cells and extracellular matrix, and fibroblasts are main cells of the dermis, and the normal proliferation and differentiation of the fibroblasts maintain the structure of the dermis of the skin and form normal structure and physiological functions; extracellular matrix is a kind of macromolecular substance synthesized and secreted by cells and interacting around the cells, and many life phenomena such as proliferation and differentiation of cells, cell migration and the like are closely related to the extracellular matrix. The extracellular matrix is composed of four major classes of molecules: collagen, elastin, glycoprotein and proteoglycan. The extracellular matrix is finely and skillfully entangled and interwoven through different molecules to form various exquisite three-dimensional structures, so that the skin is endowed with elasticity and tensile strength, and looks plump and full of vitality.

SIRT1(sirtuin 1): one of the famous longevity genes, SIRT1, is a highly conserved protein deacetylase, which participates in regulating longevity caused by energy restriction and is called a longevity gene; the SIRT1 has complex physiological action, participates in regulation of glycolipid metabolism, inflammation, oxidative stress, cell aging, apoptosis and the like, and is closely related to skin aging.

Fibroblast growth factor 7 (FGF 7) gene encodes a Fibroblast growth factor 7 mitogenic and cell survival activity and is involved in a variety of biological processes, including embryonic development, cell growth, morphogenesis, tissue repair, tumor growth and invasion. The protein is an effective epithelial cell specific growth factor, and the mitogenic activity of the protein is mainly expressed in keratinocytes, so that the proliferation rate of the skin is increased, the renewal capacity of the skin is improved, and the protein has an anti-aging effect.

The heparin binding epidermal growth factor hbegf (heparin binding EGF like growth factor) is a member of the EGF family that binds to and activates EGF receptor growth factors. HBEGF plays a key role in pathophysiological processes, including cancer. After the partial thickness of human skin is damaged, epidermal growth factor is generated to promote cell proliferation and be beneficial to healing of skin wounds, and meanwhile, EGF is the epidermal growth factor to promote the proliferation of epidermal cells and maintain the young state of the skin.

NID1(Nidogen) gene encodes protein nestin 1, which is important protein of basement membrane and plays a role in connecting type iv collagen and laminin 5, and is essential for the structure and function of basement membrane. Whether the structure and function of the basement membrane are normal or not determines whether the skin can be normally renewed and differentiated, the nestin expression is reduced, the basement membrane structure is damaged, the proliferation and differentiation of epidermal cells are influenced, and the skin is aged.

Fibrillin 1 encoded by FBN1(Fibrillin 1) gene is secreted by fibroblasts of dermis into extracellular matrix, and can be used as structural component of calcium-binding microfibril to provide stress structure for elastic and inelastic connective tissues of whole body and mediate a series of cell and matrix interactions. Is an important component of elastic fiber in connective tissue, in the process of forming the elastic tissue, the fibrillin 1 provides a skeleton for storing the elastin, the periphery of the mature elastic fiber grows, the reduction of the skin elasticity is an important characteristic of skin aging, the expression level of the fibrillin 1 is closely related to the skin elasticity, and the reduction of the expression level of the fibrillin 1 is also a key factor causing the skin aging.

Fibronectin, also known as Fibronectin, encoded by the FN1(Fibronectin 1) gene, primarily mediates cell adhesion. Purified fibronectin enhances cell-to-cell adhesion and cell-to-matrix adhesion. By adhesion, fibronectin can regulate the shape of cells and the organization of cytoskeleton through cell signal transduction pathways, promote cell spreading, promote cell growth, and make cell growth inseparable from skin state, and high expression of fibronectin can promote cell growth, so that the skin can be continuously renewed, and can resist aging.

Therefore, the research on how to improve the expression of genes related to key parts and functions of the skin has important significance for improving the stability of the true epidermal structure of the skin and delaying skin aging.

Flos Buddlejae (Buddleja officinalis Maxim.), and name of the Chinese medicinal materials. Also named: the Chinese herbal medicine is prepared from rice dyeing flower, murraya paniculata, broccoli, mongolian butterflybush flower, yellow rice flower, pimple bark tree flower and canton love-pea vine flower, is shrub of buddleia of Loganiaceae and deciduous shrub, and is 1-3 m high. The small branches are gray brown, slightly quadrangular, and densely covered by gray-white villi. The leaf pair is in a shape of a needle, the shape is narrow oval to linear, the length is 5-15 cm, the width is about 3 cm, the tip is tapered, the base is wedge-shaped, the whole edge is provided with small saw teeth, the surface is provided with fine star-shaped hair, the veins are sunken, the lower part is densely provided with grey white to yellow star-shaped hairy hair, and the veins are raised; the petiole is 6-10 mm long and is hairy with grey white; the leaves are withered at the base parts of the two petioles to form a transverse line, the panicle grows at the top of the panicle, grows for 5-12 cm, is densely covered with grey-white soft hair, and the bracts are coated with needles and are covered with villi; calyx is bell-shaped, and the tip is 4 cleft; the corolla is cylindrical, the length is about 1.5 cm, the tip is 4 cracks, the drum is violet, the mouth is orange, and the inside and the outside are soft hair; stamen 4, which grows in the middle of the corolla tube; the ovary is superior, 2 rooms, the tip is hairy, the style is short, and the stigma is not cracked. The capsule is 2-6 mm long, 2 petals are split, and the base part has the reserved calyx and petals. Most seeds are fine and flat. The flowering period is 2-3 months. The fruit period is 7-8 months, is dry flower or flower bud of Buddleja officinalis of Loganiaceae, has effects of dispelling pathogenic wind, cooling blood, moistening liver, and improving eyesight, and can be used for treating conjunctival congestion, swelling and pain, lacrimation, photophobia, cataract, and blepharitis. The 2800 m old shrub cluster or forest edge on sunny hillside, river side, village with elevation of 200 is distributed in China, Plumbum preparatium, Burma, Vietnam, mainly produced in Hubei, Sichuan, Shaanxi, Henan. In addition, it is also produced in Yunnan, Guangxi and Hunan provinces. .

The butterflybush flower was first documented in "Kaibao Bencao" in Song Dynasty, and mentioned in "Bencao Jing Shu" in Ming Dynasty: "Buddleja officinalis, who watches the book of Ben Jing (book of academic records), does not have the symptoms of liver deficiency and heat, covers the liver to open orifices, and eyes can see the blood, and liver blood deficiency indicates the symptoms of cyanosis, nebula, severe liver heat, red swelling, gum secretion, red pulse, infantile bean sore, residual toxicity, and malnutrition. The herb is sweet and effective in tonifying blood, cold in removing heat, and liver blood is sufficient, so that the symptoms are not cured. Mention is made in "materia medica truth of the Qing Dynasty": pale butterflybush flower, sweet and slightly cold … … enters liver meridian to remove heat and nourish nutrient. The liver covers the eyes to induce resuscitation, blood can be seen, skin is bluish and nebula is seen in deficiency, red and swollen lacrimation is seen in heat. The eyes with red veins, the infantile acne and residual toxicity, malnutrition and qi attacking the eyes, so it is sweet in tonifying and cold in removing heat, and liver blood and foot are not healed . However, the flavor is thinner than qi, so it is more powerful with the action of nourishing blood. The flos Buddlejae water extract and its flavonoid monomer components have inhibitory effect on Staphylococcus aureus and beta hemolytic streptococcus, and the verbascoside in flos Buddlejae also has antifungal activity. Luteolin, verbascoside, cyclamate and poliumoside separated from flos Buddlejae have antioxidant activity in vitro. The main effective components and pharmacological research progress of the buddleja officinalis show that the buddleja officinalis has certain research value and application potential in the field of skin external application, including the aspects of improving skin vitality, resisting radiation damage and the like.

Disclosure of Invention

The invention aims to solve the technical problem that substances for improving the anti-aging capacity of skin are lacked in the prior art, and provides application of a buddleja officinalis alcohol extract. The alcohol extract of the buddleja officinalis can improve the expression of genes related to skin aging resistance, and further has the function of delaying the aging of human skin.

the invention firstly provides that the alcohol extract of the buddleja officinalis has the capability of improving the expression of extracellular matrix genes, basement membrane related genes and anti-aging related genes, stabilizes the skin structure, regulates and controls the synthesis of elastic fibers in dermis, and is suitable for preparing the skin external preparation with the effect of delaying aging.

The invention mainly solves the technical problems by the following technical means:

The invention provides application of alcohol extract of buddleja officinalis in preparation of an expression up-regulator of anti-skin-aging genes.

The anti-skin aging gene is not limited to genes directly related to skin aging, such as genes involved in skin tissue damage repair and fibrosis processes, but also relates to genes indirectly related to skin aging, such as basement membrane related genes, the expression of which is improved, so that the stability of a basement membrane structure is promoted, the tight connection of the true epidermis is strengthened, and the skin aging is resisted. The skin aging resistant gene is preferably one or more of SIRT1 gene, FGF7 gene, HBEGF gene, NID1 gene, FBN1 gene and FN1 gene.

The alcohol extract of the buddleja officinalis can be alcohol extract of the buddleja officinalis which is conventional in the field, and preferably alcohol extract of the buddleja officinalis from C1 to C5; more preferably, the buddleja officinalis is an ethanol extract, and the ethanol is preferably 70-95% ethanol;

The alcohol extract of buddleja officinalis is preferably prepared by a method comprising:

(1) Pulverizing flos Buddlejae to obtain powder; the flos Buddlejae is preferably flos Buddlejae whole plant;

(2) Adding ethanol for extraction, and filtering to obtain filtrate; the extraction temperature is preferably room temperature, and the room temperature is 20-30 ℃;

(3) Evaporating the filtrate to obtain; the degree of evaporation is to dryness.

In order to more effectively extract the effective components in the buddleja officinalis: the mesh number of the powder in the step (1) is 20-60 meshes; the ethanol in the step (2) is preferably 70% ethanol, and the addition amount of the ethanol is preferably 8-12 times, more preferably 10 times of the volume of the crushed butterflybush flower; the number of times of extraction in step (2) is preferably 1 or more, more preferably 2; wherein the extraction is preferably ultrasonic extraction, and the frequency of the ultrasonic is preferably 400-550W, more preferably 500W; the ultrasonic treatment time is 25-40min, preferably 30 min.

The filtration method in step (2) is centrifugation, the centrifugation speed is preferably 6000-10000rpm, more preferably 8000rpm, and the centrifugation time is preferably 8-12 minutes, more preferably 10 minutes.

In a preferred embodiment of the present invention, the alcohol extract of buddleja officinalis is prepared by a method comprising:

(1) pulverizing dried whole plant of flos Buddlejae to obtain powder with mesh number of 20;

(2) Adding 70% ethanol with a volume 10 times of the powder, performing 500W ultrasonic extraction at room temperature for 2 times, each for 30min, centrifuging, filtering, and mixing filtrates; the speed of the centrifugation is 8000rpm, and the time of the centrifugation is 10 minutes;

(3) And (5) rotatably evaporating the filtrate to dryness to obtain the product.

The concentration of the alcohol extract of the buddleja officinalis prepared by the method in the expression up-regulator of the anti-skin-aging gene is preferably 0.8-12 mu g/ml; more preferably 1 to 10. mu.g/ml; wherein, the concentration in the expression up-regulator for HBEGF gene is preferably 0.8-1.2 μ g/ml, more preferably 1 μ g/ml; the concentration in the up-regulator for expression of FGF7 and/or FN1 gene is preferably 8 to 12. mu.g/ml, more preferably 10. mu.g/ml.

The expression up-regulator of the anti-skin aging gene is suitable for cosmetics or skin care products.

unless otherwise specified, all references to "extract of Buddleja officinalis" in the present invention refer to "alcohol extract of Buddleja officinalis".

On the basis of the common knowledge in the field, the above preferred conditions can be combined randomly to obtain the preferred embodiments of the invention.

The reagents and starting materials used in the present invention are commercially available.

The positive progress effects of the invention are as follows:

the alcohol extract of flos Buddlejae can promote synthesis of extracellular matrix, stabilize basement membrane structure, stabilize skin structure, induce cell repair-related and anti-aging-related gene expression, and improve skin structure stability and antiaging effect; under a specific concentration range, the expression level of the skin anti-aging related gene can be up-regulated to more than 1.36 times, and can reach 2.13 times at most. The discovery is of great significance for developing skin care products or cosmetics related to anti-aging in the future.

Drawings

FIG. 1 is a graph showing the results of 1. mu.g/mL of the extract of Buddleja officinalis promoting the expression of FBN1 gene.

FIG. 2 is a graph showing the results of the promotion of FBN1 gene expression by the extract of Buddleja officinalis at 10. mu.g/mL.

FIG. 3 is a graph showing the results of FN1 gene expression promotion by 10. mu.g/mL Buddleja officinalis extract.

FIG. 4 is a graph showing the results of 1. mu.g/mL of a Buddleja officinalis extract promoting SIRT1 gene expression.

FIG. 5 is a graph showing the result of 1. mu.g/mL of the extract of Buddleja officinalis promoting HBEGF gene expression.

FIG. 6 is a graph showing the results of 10. mu.g/mL of a Buddleja officinalis extract promoting SIRT1 gene expression.

FIG. 7 is a graph showing the results of 10. mu.g/mL of the extract of Buddleja officinalis promoting the expression of FGF7 gene.

FIG. 8 is a graph showing the results of 1. mu.g/mL of the extract of Buddleja officinalis promoting the expression of NID1 gene.

FIG. 9 is a graph showing the results of using 10. mu.g/mL of the extract of Buddleja officinalis to promote the expression of NID1 gene.

Detailed Description

The invention is further illustrated by the following examples, which are not intended to limit the scope of the invention. The experimental methods without specifying specific conditions in the following examples were selected according to the conventional methods and conditions, or according to the commercial instructions.