CN111803420B

CN111803420B - Cosmetic composite with anti-pollution heavy metal resistance

Info

Publication number: CN111803420B
Application number: CN202010800429.4A
Authority: CN
Inventors: 金荣熙; 金延埈; 周春亚; 曹铉; 金庚泰; 李载雨; 刘畅; 刘培玉; 冯菊; 王洁; 庞雪
Original assignee: Hyundai Bioland Jiangsu Co Ltd; Cosmax China Cosmetics Co Ltd
Current assignee: Hyundai Bioland Jiangsu Co Ltd; Cosmax China Cosmetics Co Ltd
Priority date: 2020-08-11
Filing date: 2020-08-11
Publication date: 2023-01-31
Anticipated expiration: 2040-08-11
Also published as: CN111803420A

Abstract

The invention discloses a cosmetic compound with the effects of resisting pollution and heavy metals, which is prepared by compounding burdock and water shield, effectively inhibits the stimulation of environmental harmful factors to the skin, reduces the skin injury and recovers the barrier function of the skin. The complex is a product compounded by natural plants, has mild texture, easy absorption and no stimulation, meets the requirements of people on the safety and effectiveness of cosmetics, and has wide application prospect.

Description

Cosmetic compound with anti-pollution and heavy metal-resistant functions

Technical Field

The invention relates to the field of cosmetics, in particular to a cosmetic compound with pollution resistance and heavy metal resistance.

Background

The human skin is used as a body part directly exposed to the external environment, is a protective film for protecting important organs of the human body, and plays a role in regulating water evaporation and protecting the body from external infection. However, excessive ultraviolet rays, contaminated environments, etc. induce skin irritation, resulting in skin aging.

Among such skin irritants, fine dust, yellow sand, and haze mixed with harmful substances such as heavy metals, which have been induced by recent industrial development, are the main causes of skin aging and skin problems. The heavy metal refers to mercury, cadmium, lead, copper and the like with the density of more than 4g/cm ³ Generally, the metal(s) of (1) is (are) incorporated into a substance in the human body to form an organic complex which is not easily decomposed when inhaled, and is not rapidly discharged from the body, and is accumulated on internal organs such as the liver and kidney, or bones. Long-term heavy metal inhalation can cause symptoms of quadriplegia, dyskinesia skin pigmentation and keratinization, toenail, heel and hair atrophy, reproductive dysfunction, abnormal growth, hypoevolutism, and immunologic hypofunction. Environmental hazardous substances are considered one of the most harmful environmental problems to health, and have a great correlation with skin diseases (environmental risk assessment and regulation research on heavy metal pollution [ D)]University of Nanjing, 2011.). Harmful heavy metal dust, etc. chemically react in the atmosphere, and in addition, harmful substances such as Nitrogen Oxides (NO), sulfur Oxides (SO), etc. are generated, increasing inflammatory substances of the skin, and possibly causing skin problems such as allergy, etc. The particle size of heavy metals is 2 \13211andbelow, and is much smaller than that of general dust (average size is 20 \13211andabove). The fine dust containing the heavy metals can easily enter the deep layer of skin pores because of small particles, and skin inflammation and skin sensitivity are easily caused if the fine dust invading into the skin can not be cleaned.

Clinically, the harmful substance direct contact of environment to skin can arouse anaphylactic reaction and stimulation, produces contact dermatitis easily, and simultaneously, the moisture that skin can be taken away to dry awful yellow sand, arouses skin dryness, cutin, itch, yellow sand pox and anaphylaxis etc. these problems are not uncommon, but corresponding treatment and countermeasure are still not clear and definite. Heavy metals, fine dust, yellow sand and the like, which are harmful substances, are considered as stresses in the skin, and these stresses are generated under the influence of free radicals. Free radicals (Free physics) refers to atoms or groups with unpaired electrons that lose a pole upon interaction with other molecules, which are in a very unstable state, to "treat" themselves, and to rob a pole from surrounding healthy molecules (korean journal of skin cosmetology 7 (2): 51-62). In this process, more free radicals are formed and the tissue is damaged. Since such radicals are easily reactive with living bodies, if they are not removed from the human body, oxidative stress (oxidative stress) is induced, leading to collagen and the like cleavage and an increase in MMP-1 activity, and finally, reduction in skin moisture and accelerated aging.

Disclosure of Invention

In order to solve the above problems, the present invention discloses a cosmetic composition having anti-contamination and heavy metal resistance.

The technical scheme of the invention is as follows: a cosmetic composite with anti-pollution and heavy metal resistance comprises burdock and water shield, and the raw material formula of the composite comprises the following components:

1 to 10 portions of burdock

1 to 10 parts of water shield.

Further, the proportion of the burdock to the water shield is 1: 10-10: 1.

preferably, the proportion of the burdock to the water shield is 1:1.

furthermore, the compound containing burdock and water shield has an inhibiting effect on the AhR and CYP1A1 genetic factor expression increase caused by the stimulation of an environment harmful factor benzopyrene.

The invention has the advantages that: research on natural substances capable of protecting skin from being damaged by benzopyrene, one of representative environmentally harmful substances causing skin irritation, and 2 kinds of heavy metals (arsenic and cadmium) has been conducted, and the burdock and water shield compound can be used as a cosmetic raw material for relieving and protecting skin damage by protecting skin keratinocytes at the outermost layer of the skin from being invaded by external harmful factors. The compound has an inhibiting effect on AhR and CYP1A1 genetic factor expression increase caused by the stimulation of an environmental harmful factor benzopyrene, can improve skin damage caused by heavy metal, thereby recovering the normal protection effect of the skin, and is easy to be used as a cosmetic raw material for resisting pollution, improving skin damage and protecting the skin.

Drawings

FIGS. 1-7 show the results associated with cytotoxicity of the present invention;

FIGS. 8-9 are graphs of cell viability in accordance with the present invention;

FIGS. 10 to 11 are graphs showing the expression of the genetic genes of the present invention.

Detailed Description

For the purpose of enhancing the understanding of the present invention, the following detailed description of the present invention will be given in conjunction with examples and drawings, which are provided for the purpose of illustration only and are not intended to limit the scope of the present invention.

Adding 10 times of 70% (w/w) ethanol water solution into dried burdock and water shield, extracting at 60 deg.C for 5 hr, sieving with 250 mesh sieve, and filtering with 0.3 v/13211a filter paper plate. The filtrate was concentrated under reduced pressure using a vacuum concentrator (EYELA, N-1000, japan) to obtain dry powders as control 1 and control 2. The raw materials were mixed in accordance with the following table, and examples 1 to 5 were prepared in accordance with the above-described method.

Cytotoxicity evaluation test of the Complex

In order to evaluate toxicity in the cells of the 2 control groups and 5 examples, the cell viability was determined by the MTT assay method. MTT assay is an experimental method widely used in the detection of cytotoxicity or cell proliferation by measuring the number of living cells, and is characterized in that succinate dehydrogenase (or mitochondrial dehydrogenase) in mitochondria of living cells can reduce water-soluble yellow salt MTT (3- [4, 5-dimethylthiozolin-2-yl ] -2, 5-diphenyltetrazolium bromide) into water-insoluble blue color and deposit in cells, while dead cells do not have such a function. The generated crystalline formazan (formazan) is dissolved in DMSO (dimethlysulfoxide) and then the absorbance is measured. Specifically, a 24 well multi plate (corning) was inoculated with 1ml each of 10% bovine serum-containing DMEM medium and keratinocyte (HaCaT) at a density of 1.5X105 cells/well, and the medium was changed to a serum-free medium after 24 hours of culture. Samples with different concentrations were added to serum-free medium for 24 hours after treatment. Thereafter, the medium was removed, treated with 0.25 mg/ml MTT solution, and allowed to react at 37 ℃ for 4 hours. The MTT crystalline formazan was dissolved by adding 1ml of DMSO to the cells from which the MTT solution was removed, and the absorbance was measured at 570nm, and the cell viability was calculated according to the following formula.

The control group was tested without the addition of sample. The results related to cytotoxicity are shown in tables 1 to 2 and FIGS. 1 to 7.

[ TABLE 1 ]

[ TABLE 2 ]

And (4) conclusion: the cell survival rate of the control group 1, the control group 2 and the examples 1 to 5 is close to 100 percent at all treatment concentrations of 1 to 250ug/ml, and toxicity does not occur.

Efficacy test of the Compound for protecting cells against heavy metals

In order to evaluate the effect of protecting cells against heavy metals, the degree of recovery of cell viability is determined for 2 representative heavy metals arsenic (As) and cadmium (Cd) contained in the dust particles. The efficacy of protecting cells against heavy metals is obtained by MTT assayMTT assay is an assay widely used for detecting cytotoxicity or cell proliferation by measuring the number of living cells, and the detection principle is that succinate dehydrogenase (or mitochondrial dehydrogenase) in mitochondria of living cells can make MTT (3- [4, 5-dimethylthiozole-2-yl) a water-soluble yellow salt]2,5-diphenyl tetrazolium bromide) to a water insoluble blue color and deposited in the cells, whereas dead cells did not have this function. The generated crystalline formazan (formazan) is dissolved in DMSO (dimethlysulfoxide) and then absorbance is measured. The specific experimental procedure was as follows 1.5X10 in a 24 well multi plate (corning) ⁵ cells/well were inoculated at a density of 1ml each with DMEM medium containing 10% bovine serum and keratinocytes (HaCaT), and 24 hours later, the medium was changed to serum-free medium. Heavy metals arsenic (As) and cadmium (Cd) are added into a serum-free culture medium, and samples with different concentrations are respectively treated and then cultured for 24 hours. Thereafter, the medium was removed, treated with 0.25 mg/ml MTT solution, and allowed to react at 37 ℃ for 4 hours. DMSO was added to the cells from which the MTT solution was removed, MTT crystalline formazan was dissolved, and the absorbance was measured at 570nm, and the cell viability was calculated according to the following formula.

The results are shown in tables 3 to 4 and FIGS. 8 to 9.

[ TABLE 3 ]

[ TABLE 4 ]

As a result: different samples are used for 2 heavy metals CdCl ₂ , As ₂ After the O3 is treated, the cell survival rate is increased, and compared with the single burdock or water shield extract, the effect of mixing the burdock and the water shield extract is betterGood, and example 3 is the best in protecting cells from harmful factors such as heavy metals.

Testing of the anti-fouling efficacy of the composite

To evaluate the efficacy against contamination, benzopyrene (Benzopyrene) of PAHS, a representative environmental contamination mutagen, which has an effect on DNA damage, inflammation, ROS production, and MMP-1 activity, was used as a contamination source. Thus, contaminants due to benzopyrene are produced, inflammation and aging are induced by increasing the expression of Ahr (Aryl Hydrocarbon Receptor) and inflammatory cytokine (IL-1 α) which act as environmental and chemical stimuli and immune responses to link the environment, and the expression of CYP (Cytochrome P450) 1A1, which is one of enzymes required for metabolism of foreign substances out of the body, is increased, and an aging state under a harmful environment is produced, and then the anti-pollution efficacy is determined by effectively reducing Ahr, CYP1A1, and IL-1 α genetic factors. The specific experimental method was as 1.0X10 in 6 well Multi plate (corning) ⁶ cells/well were inoculated at a density of 2ml each with 2ml of a DMEM medium containing 10% bovine serum and keratinocytes (HaCaT), and cultured for 24 hours. One day later benzo (a) pyrene (Wako, 020-13591) was added to 2ml serum-free medium in CO ₂ Incubate in the incubator for 30 minutes. After removal of the medium, 2ml of fresh serum-free medium was added, the sample was added, and the mixture was incubated with CO ₂ Incubated in an incubator for 4 hours. QIAzol was treated in cultured cells after washing with PBS buffer ^® (QIAGENL ^® USA), the manufacturer (QIAGENL) was used after cell lysis ^® ) The procedures provided isolate RNA. Using Qubit ^® The isolated RNA \47484wasquantified by fluoroemeter with RNA BR Assay kit, followed by cDNA synthesis, and Real-time fluorescence quantification (Real-time PCR) was performed. cDNA Synthesis Kit (qPCRBIO cDNA Synthesis Kit, PCRBIOS, london, UK) was used for cDNA Synthesis, and experiments were performed according to the Kit method. Real-time fluorescent quantitation (Real-time PCR) the genetic factor amplification product was quantitatively analyzed using the Real-time PCR Kit (2 x qPCRBIO SyGreen Blue mix Lo-ROX, PCRBIOS, london, UK).

Ahr, cyp1A1, IL-1. Alpha., beta. -Actin primers used in PCR were synthesized by cosmogenetech (Korea), and the primer sequences are shown in Table 5.

[ TABLE 5 ]

Primer and method for producing the same

The cDNA synthesis reaction conditions were Ahr, cyp1A1, IL-1. Alpha. And. Beta. -Actin were all reacted at 42 ℃ for 30 minutes and 85 ℃ for 10 minutes. The positive control group used Resveratrol (Sigma, R5010), a well-known antioxidant. The results of the gene expression are shown in tables 6 to 7 and FIGS. 10 to 11.

[ TABLE 6 ]

[ TABLE 7 ]

As a result: resveratrol, used as a positive control, is an AhR inhibitor and an antioxidant to prevent CYP1A1 transcription as one of the phytoalexins. Compared with Resveratrol (Resveratrol) as a positive control group, the burdock extract and water shield extract alone and all the examples showed better inhibition capability to AhR and CYP1A1 expression, of which example 3 showed the most excellent inhibition capability to AhR and CYP1A1 expression.

Claims

1. A cosmetic composition having anti-fouling heavy metal resistance, characterized by: the compound is extracted from burdock and water shield, and the raw material formula of the compound comprises the following components:

1 to 10 parts of burdock;

1 to 10 parts of water shield;

the preparation method of the compound comprises the following steps: preparing dried burdock and water shield, adding 10 times of 70% (w/w) ethanol water solution, extracting at 60 deg.C for 5 hr, sieving with 250 mesh sieve, filtering with 0.3 v/13211a filter paper plate, and concentrating the filtrate under reduced pressure with a reduced pressure concentrator to obtain dried powder.

2. The cosmetic composition with anti-contaminant heavy metal resistance of claim 1, wherein: the proportion of the burdock to the water shield is 1: 10-10: 1.

3. the anti-contaminant heavy metal resistant cosmetic composition of claim 2, wherein: the proportion of the burdock to the water shield is 1:1.

4. the cosmetic composition of claim 1, wherein the anti-contaminant heavy metal-resistant cosmetic composition comprises: the compound has an inhibition effect on AhR and CYP1A1 genetic factor expression increase caused by the stimulation of an environmental harmful factor benzopyrene.