CN111658682A - Application of passionflower flower extract in promoting skin cell mitochondrion activity and anti-aging gene expression - Google Patents
Application of passionflower flower extract in promoting skin cell mitochondrion activity and anti-aging gene expression Download PDFInfo
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Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
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- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
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- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
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Abstract
The invention relates to the field of plant extracts, in particular to application of a passion flower extract in promoting mitochondrial activity of skin cells and anti-aging gene expression. The invention provides application of passion flower extract in preparing a composition for promoting mitochondrial activity of skin cells or anti-aging gene expression. The passion flower extract not only helps to increase cellular energy production, but also improves the expression of various anti-aging genes related to mitochondrial renewal, cell stress response, and telomere elongation, such as PINK1, ATG1, ATG8, SIRT1, FOXO3, TERT, and TERC, thereby maintaining the vitality of skin cells and the youthful state of skin.
Description
Technical Field
The invention relates to the field of plant extracts, in particular to application of a passion flower extract in promoting mitochondrial activity of skin cells and anti-aging gene expression.
Background
With the increase of the average age of the population, the elderly living with good health and high quality becomes one of the focuses of public attention, and scientific research on aging is also receiving more attention. Aging can be broadly defined as a decline in physiological function over time, which is associated with a number of prevalent diseases, such as cardiovascular disease, cancer, metabolic disease, neurodegenerative disease, and the like. The aging process is complicated and influenced by many factors, including diet, exercise, mental state, and genetic factors. At the molecular or cellular level, possible internal mechanisms include telomere (telomere) shortening at the ends of chromosomes, gene mutations, gene expression disorders, constant loss of protein, decreased mitochondrial function, and the like. In addition, free radicals (e.g., reactive oxygen species) generated by physical or chemical factors in the surrounding environment of the cell damage the chromosomal deoxyribonucleic acid (DNA), proteins, and lipid biofilms of the cell, thereby disrupting the function of the cell.
Given the complexity of the factors responsible for aging, delaying the body and even the aging of various cells can be initiated from a variety of aspects, including altering dietary and motor habits, maintaining mood, reducing exposure to environmental factors that induce the formation of free radicals (e.g., ultraviolet light), and enhancing the functioning of cytoprotective mechanisms (e.g., promoting the expression of anti-aging genes). Among them, methods for promoting the cell protection mechanism are in the development stage, and the means for enhancing the cell protection mechanism are under study. Therefore, there is a need to develop a novel composition that can promote the cell protection mechanism, and provide a new direction for slowing down the cell function decline and maintaining the cell health status.
Disclosure of Invention
Accordingly, an object of the present invention is to provide a use of a passion flower (Passiflora spp.) extract for preparing a composition for promoting mitochondrial activity in skin cells, wherein the passion flower extract is obtained by extracting a passion flower with a solvent.
In one embodiment of the present invention, the passionflower flower extract is extracted with water, alcohol, or a mixture of alcohol and water as a solvent.
In one embodiment of the present invention, the passion flower extract inhibits the expression of a poly (adenosine diphosphate ribose) polymerase (also known as poly (ADP-ribose) polymerase, PARP), and the poly (adenosine diphosphate ribose) polymerase includes poly (adenosine diphosphate ribose) polymerase 1(PARP1) and poly (adenosine diphosphate ribose) polymerase 2(PARP 2). Since PARP is known to be associated with mitochondrial dysfunction, the extract of passion flower may, but is not limited to, enhance mitochondrial activity by inhibiting PARP gene expression.
Another object of the present invention is to provide a use of a passion flower extract for preparing a composition for promoting the expression of skin cells anti-aging genes, wherein the passion flower extract is obtained by extracting a passion flower with a solvent.
In one embodiment of the present invention, the anti-aging gene encodes an autophagy-related protein (ATG), including autophagy-related protein 1(ATG1) and autophagy-related protein 8(ATG 8).
In one embodiment of the invention, the anti-aging gene encodes a phosphatase and tensin homolog induced kinase 1(PINK 1).
In one embodiment of the present invention, the anti-aging gene encodes a sirtuin 2homolog 1(SIRT1) or a forkhead box protein O3(forkhead box class O3, FOXO 3).
In one embodiment of the invention, the anti-aging gene encodes a telomerase reverse transcriptase (TERT), a telomerase RNA component (TERC), or a combination thereof. Therefore, the passion flower extract can promote the synthesis of telomerase in skin cells.
In one embodiment of the present invention, the concentration of the passion flower extract for promoting mitochondrial activity or anti-aging gene expression in skin cells is 0.125mg/mL to 0.25 mg/mL.
Based on in vitro cell experiments and gene expression analysis results, the invention discloses that the passion flower extract can improve the mitochondrial activity of skin cells and the expression of various anti-aging genes. In view of the fact that maintenance of mitochondrial activity is very important for cell viability, and the anti-aging genes are involved in events that are conducive to cell rejuvenation, such as aged mitochondrial turnover, cell stress response (i.e., various molecular mechanisms that assist in reducing cell damage in stress), and lengthening of telomeres, passionflower flower extract has the potential to maintain skin cell viability and the young state of individual skin. Accordingly, the present invention provides the use of a passion flower extract for the preparation of a composition for promoting mitochondrial activity in skin cells or anti-aging gene expression. The composition can be in the form of powder, granule, liquid, gel or paste, and can be made into food, beverage, medicine, reagent or nutritional supplement, and administered to an individual by oral administration, skin application, etc.
The following embodiments are provided to illustrate the features and applications of the present invention, rather than to limit the scope of the invention, and it will be apparent to those skilled in the art that various changes and modifications can be made therein without departing from the spirit and scope of the invention.
Drawings
FIG. 1 shows the relative amount (%) of JC-1 aggregates in human dermal fibroblasts 24 hour after treatment with or without Passiflora flower extract;
FIG. 2 shows the relative expression of PARP1 and PARP2 genes in human skin fibroblasts 48 hours after treatment with and without Passiflora flower extract relative to control cells;
FIG. 3 shows the relative expression of the PINK1, ATG1, ATG8, SIRT1, FOXO3, TERT, and TERC genes in human skin fibroblasts 48 hours after treatment with and without Passiflora flower extract relative to control cells.
Detailed Description
The invention provides application of passion flower extract in preparing a composition for promoting mitochondrial activity of skin cells or anti-aging gene expression. The passion flower extract of the invention is obtained by extracting passion flower with a solvent, wherein the solvent can be water, alcohol or alcohol-water mixture, preferably water. The following examples show that treatment of the passion flower extract can increase mitochondrial activity in human skin cells and inhibit gene expression of various poly ADP ribose polymerases (PARP; e.g., PARP1 and PARP2) associated with mitochondrial dysfunction. In addition, the passion flower extract promotes the expression of anti-aging genes in human skin cells. The products expressed by the anti-aging genes may be ribonucleic acids (RNA) or proteins, including but not limited to: phosphatase and tensin homolog inducible kinase 1(PINK1), various autophagy-related proteins (ATG; e.g., ATG1 and ATG8), Sirtuin 2homolog 1(SIRT1), forkhead box protein O3(FOXO3), and telomerase reverse transcriptase (TERT) and telomerase RNA component (TERC) involved in telomerase reactions.
Definition of
As used herein, the numerical values are approximations and all numerical data are reported to be within the 20 percent range, preferably within the 10 percent range, and most preferably within the 5 percent range.
As used herein, "mitochondrial activity" refers to the efficiency of mitochondrial operations, including energy production, metabolism, efficiency of active oxygen species scavenging, and the like. Assessment of mitochondrial activity can be accomplished according to biochemical, molecular biological, and cytological techniques well known in the art, such as the measurement of mitochondrial membrane potential as described herein.
The term "anti-aging gene" as used herein generally refers to a gene whose presence is associated with the longevity of an organism or whose action of RNA or protein products maintains the normal function of a cell. The anti-aging gene encodes proteins which have the functions of promoting normal mitochondrial operation, assisting folding of other proteins, promoting telomerase synthesis, regulating gene expression of proteins involved in nutrient metabolism or cell stress response and the like.
Materials and methods
Material
Eagle's minimal medium (Gibco Eagle's minimal essential medium, MEM), fetal bovine serum (Gibco fetalbovene serum, FBS), sodium pyruvate, penicillin/streptomycin, and Phosphate Buffered Saline (PBS) containing Earle's Balanced Salt Solution (EBSS) were purchased from Thermo Fischer Scientific, Inc.
Cell culture
The following examples use human skin fibroblast (BCRC) CCD-966SK (BCRC 60153) purchased from Bioresource Collection and Research Center (BCRC). CCD-966SK cells were cultured in MEM medium (containing EBSS) to which 10% fetal bovine serum, 1mM sodium pyruvate, 0.01M nonessential amino acids, and 1% penicillin-streptomycin were added, hereinafter referred to as MEM cell culture medium, at 37 ℃ under 5% carbon dioxide.
Preparation of passionflower flower extract
The passion flower extract is obtained by extracting a flower of a passion plant (Passiflora spp.) with a solvent (called passion flower for short), such as passion fruit (Passiflora edulis) and passion flower (Passiflora foetida). The solvent may be water, alcohols, or alcohol water mixtures, preferably water. In one embodiment, the passionflower flowers are washed and dried before extraction, and then coarsely crushed using a homogenizer. Then, uniformly mixing the solvent and the homogeneous passionflower flower mass in a stirring manner for 0.5 to 3 hours to obtain a passionflower flower extract; the weight ratio of the solvent to the homogeneous passionflower flower mass is in the range of 20: 1 to 2: 1, preferably 5: 1; the extraction temperature is 40 ℃ to 75 ℃. The extracts of passion flower used in the following examples were obtained from jaboticaba biotechnologies, inc, and were prepared in a manner similar to the above procedure.
Mitochondrial Activity assay
To evaluate the mitochondrial activity of cells, flow cytometry (Beckman Coulter Life Sciences) and the mitochondrial membrane potential detection kit (BD)TMMitoScreen) measures changes in mitochondrial membrane potential, whichThe procedure is briefly described below. The test cells were rinsed in PBS according to the manufacturer's instructions, collected in a 1.5mL microcentrifuge tube and centrifuged (400Xg, 5 minutes). The supernatant was removed, the cells resuspended in PBS solution and centrifuged again (400xg, 5 min). 100 μ L JC-1(5, 5 ', 6, 6' -tetrachlororo-1, 1 ', 3, 3' -tetraethylene benzazolylcyanine iodide) working solution was mixed with the precipitated cells homogeneously in the dark for 15 minutes for fluorescence labeling, and the cells were washed 2 times with washing solution and centrifugation step (400Xg, 5 minutes). Finally, the cells were resuspended in 2% FBS in PBS and the relative amount of JC-1 aggregates indicative of mitochondrial membrane potential was detected using flow cytometry (excitation wavelength about 490nm, detection wavelength about 595 nm). A larger amount of JC-1 aggregate indicates a larger potential difference in the mitochondrial inner membrane and is considered to be higher in mitochondrial activity.
Analysis of Gene expression
The gene expression related to the activity of granulocytes and anti-aging in cells was determined based on quantitative polymerase chain reaction (qPCR), and the procedure is briefly described below. RNA was isolated from the cells using an RNA Extraction Kit (RNA Extraction Kit; Geneaid) according to the manufacturer's instructions and reverse transcriptase was used at 37 deg.CIII Reverse Transcriptase 2000ng of RNA was Reverse transcribed to cDNA (Invitrogen). Next, qPCR was performed on the cDNA using a qPCR Kit (KAPA CYBR FAST qPCR Kit (2X); KAPA Biosystems) and a primer set of the target genes listed in Table 1 below and Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) gene as an internal control in a PCR reactor (Step One Plus Real-Time PCR system; Applied Biosystems) to obtain a melting curve (melting curve).
TABLE 1
Finally, use 2-ΔΔCTMethod for determining target geneRelative expression of (c). The method uses the cycle threshold (C) of GAPDH geneT) The cycle threshold of the reference gene as an internal control was calculated as the relative fold change according to the following formula:
ΔCTc of target gene in experimental group or control groupTC of internal controlT
ΔΔCTΔ C of experimental groupTΔ C of control groupsT
Statistical analysis was performed by calculating the standard deviation of relative expression of each gene using the STDEV function in Excel software and calculating the statistical difference using the single-tailed student T test (TTEST).
Example 1
Passion flower extract for improving mitochondrion activity of skin cells
To investigate the effect of the passion flower extract on the mitochondrial function of skin cells, the present example uses a flow cytometer to evaluate the change in mitochondrial activity of human dermal fibroblasts, CCD-966SK, after treatment with the passion flower extract, first, the CCD-966SK cells were treated with 1 × 105Cells/well were seeded in 6-well plates, each well containing 2mL of MEM cell culture medium. After culturing the cells at 37 ℃ for 24 hours, the cell culture medium was removed and the cells were treated with 1mL of MEM cell culture medium containing 0.125mg/mL of Passiflora edulis flower extract (experimental group) or treated with MEM cell culture medium alone as a control group. Each group of cells was cultured at 37 ℃ for 24 hours and then analyzed for mitochondrial activity, with the fluorescent dye JC-1 indicating mitochondrial membrane potential and activity.
FIG. 1 shows the relative amount (%) of JC-1 aggregates in the preceding groups of CCD-966SK cells, with higher values indicating higher average mitochondrial activity of the cells;***represents p < 0.001 for the control group. According to FIG. 1, administration of 0.125% Passiflora incarnata flower extract significantly increased the relative amount of JC-1 aggregates in CCD-966SK cells by about 40% as compared to control cells, indicating that WesternThe extract of the crocus sativus can improve the mitochondrial activity of skin cells, so that the skin cells have enough activity to perform specific physiological functions or proliferate to replace old cells. Thus, the passion flower extract is beneficial for maintaining the youthful state of the skin of a human subject.
Example 2
Passion flower extract for improving cell aging resistance
To investigate the effect of passion flower extract on the anti-aging ability of skin cells, qPCR was used to determine the changes in gene expression of poly ADP ribose polymerases 1 and 2(PARP1 and PARP2), phosphatase and tensin homolog-induced kinase 1(PINK1), autophil-related proteins 1 and 8(ATG1 and ATG8), sirtuin 2homolog 1(SIRT1), forkhead box protein O3(FOXO3), telomerase reverse transcriptase (TERT), and telomerase RNA component (TERC) of human dermal fibroblast cells CCD-966SK after being treated with the passion flower extract, briefly, CCD-966SK cells were subjected to 1.5 × 105Cells/well were seeded in 6-well plates, each well containing 2mL of MEM cell culture medium. After culturing the cells at 37 ℃ for 24 hours, the cell culture medium was removed and the cells were treated with 1mL of MEM cell culture medium containing 0.125mg/mL or 0.25mg/mL of Passiflora edulis extract (experimental group) or only with MEM cell culture medium as a control group. The aforementioned cells were used for qPCR analysis after 48 hours of culture at 37 ℃.
FIG. 2 shows the relative expression of PARP1 and PARP2 genes relative to control cells after 48 hours of treatment of CCD-966SK cells with 0.125mg/mL passion flower extract. According to FIG. 2, treatment of Passiflora incarnata flower extract inhibited the gene expression of multiple Poly ADP Ribose Polymerases (PARP) in CCD-966SK cells. In view of previous studies indicating that inhibition of this enzymatic activity could improve mitochondrial activity, this result suggests that passion flower extract could increase mitochondrial activity, thereby improving cellular energy production and fighting against aging.
FIG. 3 shows the relative expression of PINK1, ATG1, ATG8, SIRT1, FOXO3, TERT, and TERC genes relative to control cells after 48 hours of treatment of CCD-966SK cells with 0.25mg/mL passion flower extract;*、**and, and***respectively representing the control group of the comparison as p < 0.05 and p <0.01, and p < 0.001. According to FIG. 3, the administration of Passiflora flower extract to CCD-966SK cells significantly increased the expression of PINK1, ATG1, and ATG8 genes associated with aged mitochondrial turnover and promoted the expression of SIRT1 and FOXO3 genes associated with cell stress response, as compared to the control group. In addition, the passion flower extract also enhanced the expression of the TERT and TERC genes involved in telomerase reactions to prolong telomeres. This result indicates that the passion flower extract enhances the production of various anti-aging genes in skin cells, thus helping to maintain the vitality of skin cells and delay skin aging.
In conclusion, the passion flower extract can improve the mitochondrial activity of skin cells and the expression of various anti-aging genes, and further maintain the vitality of the skin cells and the young state of the skin of an individual. Accordingly, the present invention provides the use of a passion flower extract for the preparation of a composition for promoting mitochondrial activity in skin cells or anti-aging gene expression. The composition can be in the form of powder, granule, liquid, gel or paste, and can be made into food, beverage, nutritional supplement, or medicinal product, and administered to an individual by oral administration, skin application, etc.
Claims (10)
1. Use of a passion flower extract for the preparation of a composition for promoting mitochondrial activity in skin cells, wherein the passion flower extract is obtained by extracting a passion flower with a solvent.
2. The use of claim 1, wherein the passion flower extract inhibits the gene expression of poly (adenosine diphosphate ribose) polymerase (PARP), and the poly (adenosine diphosphate ribose) polymerase comprises poly (adenosine diphosphate ribose) polymerase 1 and poly (adenosine diphosphate ribose) polymerase 2.
3. Use of a passion flower extract for the preparation of a composition for promoting expression of skin cell anti-aging genes, wherein the passion flower extract is obtained by extracting a passion flower with a solvent.
4. The use of claim 3, wherein said anti-aging gene encodes an autophagy-related protein (ATG) comprising autophagy-related protein 1 and autophagy-related protein 8.
5. Use according to claim 3, wherein said anti-aging gene is a gene encoding a stress protein homolog-induced kinase 1(PINK 1).
6. The use of claim 3, wherein said anti-aging gene encodes sirtuin 2homolog 1(SIRT 1).
7. Use according to claim 3, characterized in that the anti-aging gene is a gene encoding the one-forkhead box protein O3(FOXO 3).
8. The use of claim 3, wherein the anti-aging gene encodes telomerase reverse transcriptase (TERT), telomerase ribonucleic acid component (TERC), or a combination thereof.
9. Use according to claim 1 or 3, characterized in that the solvent is water, an alcohol, or an alcohol-water mixture.
10. Use according to claim 1 or 3, wherein the concentration of the passion flower extract is from 0.125mg/mL to 0.25 mg/mL.
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Citations (4)
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JP2002332224A (en) * | 2001-03-06 | 2002-11-22 | Kose Corp | Anti-ageing skin care preparation and anti-ageing skin care preparation composition |
WO2009147345A2 (en) * | 2008-05-13 | 2009-12-10 | Laboratoire Nuxe | Combination of passion flower and alkanet extracts for use in cosmetics |
WO2012172218A2 (en) * | 2011-04-20 | 2012-12-20 | Laboratoire Nuxe | Plant extract complex for skin protection |
US20180092952A1 (en) * | 2015-05-05 | 2018-04-05 | Idunn Technologies | Anti-aging composition comprising a plant extract |
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2019
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JP2002332224A (en) * | 2001-03-06 | 2002-11-22 | Kose Corp | Anti-ageing skin care preparation and anti-ageing skin care preparation composition |
WO2009147345A2 (en) * | 2008-05-13 | 2009-12-10 | Laboratoire Nuxe | Combination of passion flower and alkanet extracts for use in cosmetics |
WO2012172218A2 (en) * | 2011-04-20 | 2012-12-20 | Laboratoire Nuxe | Plant extract complex for skin protection |
US20180092952A1 (en) * | 2015-05-05 | 2018-04-05 | Idunn Technologies | Anti-aging composition comprising a plant extract |
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