Peptide bioregulators application in cosmetology: clinical and molecular aspects

Peptide bioregulators application in cosmetology: clinical and molecular aspects

The relevance of the topic. Skin aging and peptide bioregulation. The reason of accelerated skin aging is an exposure to ultraviolet radiation (UR), which is the part of the spectrum of sunlight, and stresses in everyday life. Skin aging is expressed in a decrease of its elasticity and the formation of wrinkles. Age-related changes and accelerated skin aging significantly reduce satisfaction with the appearance, especially in people who lead an active social life.

Skin aging is expressed in a violation of its endocrine, immune function and a decrease in the viability of fibroblasts — the main type of skin cells. Peptide bioregulators Endoluten^®, Sigumir^®, Cartalax^®, Crystagen^® and Vladonix^® help prevent accelerated aging and slow down the development of age-related changes in the skin.

Endoluten^® is a polypeptide complex from pineal gland, the regulator of the neuro-immuno-endocrine system and a geroprotector. The following free amino acids (3.3%), dipeptides (23.2%), tripeptides (50.7%), tetrapeptides (22.1%) and pentapeptides (0.7%) were detected in the polypeptide complex from pineal gland by mass spectrometry and high-performance liquid chromatography. The AEDG peptide was identified among tetrapeptides in pineal gland polypeptide complex by the method of selective monitoring of reactions (SRM). The biological activity inherent in pineal gland polypeptide complex is due to the effect of the AEDG peptide included in its composition [6]. The AEDG peptide regulates circadian rhythms, functions of the endocrine and immune systems, has an antioxidant effect, stress-protective effect, increases the length of telomeres in cells and life expectancy in experiments [7]. Since the skin is an organ of the neuro-immuno-endocrine system, the AEDG peptide contributes to the normalization of skin cells functions.

Sigumir^® is a polypeptide complex from cartilage and bone tissues, a regulator of connective tissue cells (cartilage and skin fibroblasts) functions. Sigumir^® was analyzed at the Institute of Toxicology of the FMBA of Russia for the content of the AED peptide (Сartalax^®) by matrix-activated laser desorption/ionization (MALDI) and ultra-efficient liquid chromatomass spectrometry (UEFC-MS). The AED peptide was found in the composition of Sigumir^® in an amount of 0.2 mg/g and is its active component [9]. Chondroprotective effects of Сartalax^® (efficacy in osteoarthritis, osteoporosis, etc.) are due to the activation of proliferation (cell division) of cartilage fibroblasts. Fibroblasts have similar properties in all organs and tissues. Thus, Cartalax^® has a stimulating effect on the metabolism of skin fibroblasts.

Vladonix^® is a polypeptide complex from thymus, a regulator of the functions of immune system. Vladonix^® was analyzed at the Institute of Toxicology of the FMBA of Russia for the content of the EDP (Crystagen^®) peptide in it by MALDI and UEFC-MS methods. The EDP peptide was found in Vladonix^® in an amount of 0.04 mg/g and is its active component [8]. The EDP peptide stimulates differentiation and activation of B-cells, macrophages, proliferation of immune cells during aging, reduces apoptosis of stem and immune cells during aging. At the systemic level, these effects of the EDP peptide are expressed in regulation of immune system functions, antioxidant action and stress-protective effect [7].

Peptide bioregulators application in cosmetology.
Middle-aged (45-59 years old) and elderly (60-74 years old) women were observed by a cosmetologist participated in the study in ArtMediA Clinic (St. Petersburg, Russia). All women gave written informed consent to participate in the study. Women were divided into equal groups of 15 people each: the 1st group (control) received a saline solution by electrophoresis on the facial skin area, once a week during 8 weeks. The 2nd group received peptides Cartalax^® and Crystagen^® (sublingual biologically active food supplements) by electrophoresis on the face area, according to the same scheme as in the control group. The 3rd group received oral polypeptide complexes (biologically active food supplements) Sigumir^® and Endoluten^® simultaneously with electrophoresis. All women excluded any cosmetic procedures 4 months before the study. The impact of skin aging on the women’s quality of life was assessed using an adapted questionnaire «Dermatological Quality of Life Index» on the first stage of the research. After that the effect of peptides on the structure of facial skin was studied using the ultrasound imaging method. The evaluation criteria were the sensations of dryness or tightness of the skin, the color from normal to dull, the surface relief of the skin, smoothness, the presence of enlarged capillaries, the severity of wrinkles, age spots and the general feeling of the appearance — freshness, radiance or tired appearance. Interpretation of results: 0 points — absence of age-related changes in all signs, 40 points — the most pronounced age-related changes in all signs. The questionnaire was filled out by women and cosmetologist. The skin structure was examined using ultrasound scanning at the next step of investigation. It was used the equipment of ultrasound diagnostics department of the E.E. Eichwald Clinic of the Nord-Western State Medical University named after I.I. Mechnikov (St. Petersburg, Russia) [1, 2].

It was found that involutes skin changes in the majority of middle-aged and elderly women significantly worsen the quality of life. After Cartalax^® and Crystagen^® application, as well as the combined use of these short peptides and Endoluten^®, Sigumir^® polypeptide complexes, subjective improvement of skin quality was noted in women of both age groups.

Application electrophoresis with Cartalax^® and Crystagen^® in combination with Endoluten^® and Sigumir^® in capsules contributed to a significant increase in the thickness of the dermis and epidermis by 15-50% in women of different ages. Ultrasound scans of middle-aged and elderly women revealed an increase in echogenicity of the dermis under the influence of the combined use of short peptides and polypeptide complexes. An increase in the echogenicity of the dermis under the influence of peptides indicates an increase in collagen synthesis.

Molecular mechanism of peptide bioregulators protective action on skin cells during their aging.

The effect of the AEDG peptide, which is part of Endoluten^®, on the expression of genes encoding enzymes of the antioxidant system in skin fibroblasts during accelerated aging. The effect of the AEDG peptide on the expression of genes encoding enzymes of the antioxidant system (NQO1, SOD1, CATALASE) in human skin fibroblasts was evaluated in an experimental model of accelerated aging induced by UR [4].

The study was used skin fibroblasts of 45 years old woman (DF2 cell line) obtained from the Collection of cell cultures of the Institute of Cytology of the Russian Academy of Sciences (St. Petersburg). Skin cells were exposed to UR during 24 hours for simulation of accelerated aging. Peptides were added to dermal fibroblasts before exposure to UR. This allowed us to evaluate the protective effect of peptides from the formation of free radicals caused by exposure to UR. Control cell cultures were grown under the same conditions without irradiation.

Superoxide dismutase 1 (SOD1) is an antioxidant enzyme that protects the intracellular space from superoxide anions by catalyzing their conversion into molecular oxygen and hydrogen peroxide. SOD1 gene expression decreases in human skin fibroblasts during aging. An increase of SOD1 gene expression in human skin fibroblasts is one of the mechanisms of response to the effects of oxidative stress (UR). In mice with a deficiency in the synthesis of SOD1 enzyme, there is a decrease in the thickness of the dermis, a violation of the migration and proliferation of fibroblasts and accelerated skin aging. The AEDG peptide increased SOD1 gene expression by 3.5 times. It is shown the protecting peptide activity on skin fibroblasts from the action of free radicals arising under the action of UR.

Catalase is an enzyme of the oxidoreductase class, which is part of the antioxidant system of the cell. It performs the function of antiperoxide protection. This enzyme catalyzes the decomposition of hydrogen peroxide formed during biological oxidation into water and molecular oxygen. The increased of catalase and SOD1 synthesis associated with aging is one of the protective mechanisms that prevent DNA damage in skin fibroblasts under the action of oxidative stress. Compensatory increase of catalase synthesis in fibroblasts was revealed with accelerated skin aging under the influence of UR. The AEDG peptide stimulates this effect by activating the antioxidant system.

NQO1 (NADP(H) — quinone oxidoreductase 1) enzyme is localized in the cytosol (inside the cell), catalyzes the two-electron reduction of quinone compounds and prevents the formation of semiquinone free radicals and active oxygen molecules, protecting cell from oxidative stress. NQO1 gene overexpression is observed in various cell types when exposed to oxidants and heavy metals. The AEDG peptide stimulates NQO1 gene expression of the antioxidant system.

Effect of the AED (Cartalax^®) and the AEDG peptides on synthesis of proteins — markers of skin fibroblasts functional activity during their aging in culture. Cartalax^® increased sirtuins 1 and 6 synthesis by 2 times and 11.5 times, and the synthesis of collagen type 1 by 2.7 times in human skin fibroblasts during their aging in culture. Collagen type 1 is one of the main structural proteins synthesized by skin fibroblasts, which indicates the high functional activity of these cells. It synthesis in fibroblasts decreases during skin aging. Sirtuin-1 (Sirt1) is a multifunctional protein that participates in stress response, cellular metabolism and aging through deacetylation of various substrates, including histones and transcription factors.

Sirtuin-1 regulates energy homeostasis, cell cycle, apoptosis, inflammatory reactions and level of reactive oxygen species in cell. Sirutin-6 (Sirt6), by weakening the action of the cytokine NFkB, inhibits cellular aging. Sirtuin-6 contributes to an increase in life expectancy. It participates in DNA repair, activates cell metabolism. The effect of Cartalax^® on Sirt1, Sirt6 and collagen type 1 synthesis is the basis of its ability to increase the functional activity of skin fibroblasts, to have an antioxidant and geroprotective effect [3].

The AEDG peptide increased Ki67 and CD98hc proteins synthesis respectively by 1.4-1.5 times in human skin fibroblasts during their aging in culture. In addition, the AEDG peptide reduced MMP9, p16, p53 and caspase-3 proteins synthesis by 1.3, 1.7, 3 and 1.4 times, respectively, in human skin fibroblasts during their aging in culture. Ki67 is one of the main proteins necessary for the proliferation (division) of skin fibroblasts. Ki67 synthesis decreases in fibroblasts during skin aging. CD98hc protein is synthesized on the membrane of skin fibroblasts and participates in their proliferation (division). CD98hc synthesis in skin fibroblasts decreases during aging, which leads to a slowdown in its regeneration. Matrix metalloproteinase-9 (MMP-9) is involved in the remodeling of the intercellular environment by skin fibroblasts. The UR-induced synthesis of MMR-9 contributes to the destruction of type 1 and 3 collagens in the dermis. The synthesis of MMP-2, MMP-3, and MMP-9 leads to the degradation of non collagenic components of the dermis, including glycoproteins and proteoglycans of the basement membrane. The level of MMP-2, MMP-3, MMP-9 tissue inhibitors decreases during aging, which contributes to the activation of remodeling of the intercellular matrix. Caspase-3 and p16, p53 transcription factors are the most important components of the cascade of apoptosis (programmed cell death) in skin fibroblasts. An increase of its synthesis and decrease of Ki67 synthesis are characteristic of skin aging [5].

Conclusion

1. Polypeptide complexes of cartilage (Sigumir^®), pineal gland (Endoluten^®), and AED (Cartalax^®), EDP (Crystagen^®), AEDG short peptides increase the thickness of the epidermis and dermis in middle-aged and elderly women according to the questionnaire and evaluation of the thickness of the epidermis and dermis by ultrasound diagnostics. This helps to slow down the aging of skin cells and improve its appearance.
2. AED (Cartalax^®) and AEDG short peptides interact with promoters of genes encoding proteins SIRT1, SIRT6, p16, p53, Caspase-3, CD98hc, MMP-9, type 1 collagen, catalase, SOD1, NQO1 — markers of skin fibroblasts functional activity. This contributes to the normalization of skin fibroblasts functions with their natural and accelerated (caused by the action of sunlight) aging.
3. The obtained data indicate the effectiveness of the use of peptide bioregulators Sigumir^®, Endoluten^®, Crystagen^® and Cartalax^® to slow down age-related skin changes. These peptide bioregulators can be used to improve the quality of life and satisfaction with their appearance in middle-aged and elderly women.

References

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