Can Cistanche Tubulosa Extract Whiten Skin?
Cistanche is a precious Chinese medicinal herb that has whitening, moisturizing, anti-aging and antibacterial effects in cosmetics and skin care. [1] Its application was first recorded in Shennong's Classic of Materia Medica, and it is second only to ginseng in the frequency of its appearance in anti-aging and antioxidant prescriptions, earning it the reputation of “desert ginseng”. [2-4] In ancient medical classics, it was recorded that the Chinese medicinal herb Cistanche deserticola can have the effect of beautifying the skin. Both the “Discussion on the Nature of Drugs” and the “Yuzhou Pharmacological Interpretation” mention that it can benefit the bone marrow, improve complexion and prolong life.
In recent years, most research has focused on the detection, identification and extraction of active ingredients in Cistanche. 70 phenylethanoid glycosides and 26 cyclic ene-ether terpenes and their glycosides have been identified, among which echinacoside and acteoside belong to the phenylethanoid glycosides. Echinacoside antioxidant and anti-aging effects, while mulberrofuran has both tyrosinase inhibitory activity and antioxidant effects, which have whitening and anti-aging effects. Both are index components specified in the 2015 edition of the Chinese Pharmacopoeia. In addition, Cistanche also contains natural polysaccharides that delay aging, resist radiation, and have antioxidant properties. This article mainly analyzes the active ingredients of Cistanche deserticola, summarizes the testing methods and extraction processes, and explains the application and efficacy mechanism of Cistanche deserticola in cosmetics, in order to expand the application of Cistanche deserticola in cosmetics.
1 The main active ingredients and testing methods of Cistanche extract's main active ingredients and detection methods
Although Cistanche has a complex and diverse chemical composition, in the study of its skin care benefits, its main active ingredients are phenylethanoids and polysaccharides. Among these phenylethanoids, echinacoside and verbascoside are the most studied. Polysaccharides polysaccharides are mainly used as anti-aging ingredients. They are composed of fructose, galactose, arabinose, ribose, glucose, rhamnose, fucose and xylose, as well as mannitol, glucuronic acid, galacturonic acid and other components [5] connected by glycosidic bonds to form straight or branched chains, and are widely used in cosmetic formulations.
With the continuous upgrading of detection technology and equipment, more and more compounds in Cistanche extract have been detected qualitatively and quantitatively. Zhou Ye et al. [6] used Fourier transform infrared spectroscopy (FTIR) combined with HPLC-ESI-MS to analyze the differences in active ingredients in Cistanche due to different production areas. Seven active ingredients such as echinacoside and verbascoside were detected in Cistanche. According to the infrared fingerprint maps of the five Cistanche herbs, the authors calculated the common peaks and difference peaks of these herbs, and then clearly analyzed the differences between the five traditional Chinese medicines Cistanche. The HPLC-ESI-MS method mainly determines the content of certain chemical components of Cistanche, with relatively high sensitivity, large sample size, and fast analysis speed. The combined analysis of the two can accurately identify the quality differences of Cistanche from different origins.
Qian Hao et al. [7] used RP-HPLC to achieve good separation of echinacoside, acteoside and other components in Cistanche deserticola. The results showed that the content of the four components in Cistanche deserticola varied greatly depending on the harvest season, providing a scientific basis for ensuring the purchase of high-quality Cistanche deserticola. Wang Xue et al. [8] used HPLC-UV, HPLC-ELSD, UV-VIS and an alcohol-soluble extract method to measure the content of echinacoside and madecassoside, galactitol and betaine, soluble polysaccharides and extract in Cistanche deserticola, to study the authenticity of Cistanche deserticola and compare the differences in various components in different seasons. Qin et al. [9] used UPLC to analyze the relative content of five phenylethanoid glycosides in Cistanche deserticola. The results showed that the content of these five components also varied depending on the place of origin.
The authors also used MALLS and ion chromatography to determine the molecular weight, distribution and monosaccharide composition of Cistanche polysaccharides, providing a methodological basis and data reference for the comprehensive evaluation of desert Cistanche. Zhu Nai et al. [10] used ultra-high performance liquid chromatography (UPLC) to demonstrate the fingerprints and differences between different types of Cistanche herbs. The results showed that there was not much difference in the content of phenylethanoid glycosides and cyclic ene-ether glycosides in Cistanche deserticola, and the content of phenylethanoid glycosides was significantly lower than that in Cistanche tubulosa. Nan Ze et al. [11] accurately identified the active ingredients in the methanol extract of Cistanche herb using HPLC-MS technology and specific chromatographic conditions. Seventeen components were identified based on retention time, relative molecular weight, and comparison of fragment ions with those of the reference substance, providing a certain standard for comprehensively controlling the quality of Cistanche herb. Research
Ren Lu et al. [13] established the adsorption and elution conditions for the separation of phenylethanoid glycosides from Cistanche deserticola by macroporous adsorption resin through static adsorption and dynamic adsorption as well as dynamic elution experiments. The AB-8 adsorption resin was selected with an adsorption capacity of 57.21 mg/g, a sample solution pH of 3.43, a sample loading concentration of 3.6–6.0 mg/L, a sample loading rate of 1.5 BV/h, and an elution with 40% ethanol aqueous solution.
Meng Qingyan et al. [14] used single-factor experiments to investigate the effects of extraction time, material-to-liquid ratio, extraction temperature, ethanol volume fraction and extraction times on the extraction process. Then, an L9(34) orthogonal design table was used to conduct orthogonal experiments on the material-to-liquid ratio, extraction times and ethanol extraction fraction. The results showed that the number of extractions had the most significant effect. Heating and refluxing at 80 °C for 2 h, a liquid-to-solid ratio of 1:20, and then extracting with 40% ethanol once, reducing the pressure until the smell of ethanol disappears, vacuum drying, to obtain crude phenylethanoid glycosides.
Zhang Kai [15] extracted phenylethanoid glycosides from Cistanche deserticola, and used Soxhlet extraction with ethanol as the solvent; extraction with ethyl acetate to degrease and n-butanol as the solvent; and distillation to dryness. Orthogonal experiments were also used to determine the significant influencing factor of the ethanol Soxhlet extraction process, i.e., the extraction time, and to optimize the best extraction process conditions.
Wang Yingzi et al. [16] used three methods to extract echinacoside, β-sitosterol, and total sugar from Cistanche deserticola. Under the same technological conditions such as the mesh size of the crushed Cistanche deserticola, the amount used, filtration, and concentration, , the flocculation water extraction method is superior to the other two methods. The yields of echinacoside, β-sitosterol and total sugar are 25.46 mg/g, 0.38 mg/g and 12.84 mg/g, respectively.
Gao Lei et al. [17] invented a patented process for the extraction of Cistanche polysaccharides using a combination of enzymes and ultrasound. The extraction process conditions were optimized using single-factor experiments and orthogonal design experiments to obtain the process with the highest yield of Cistanche polysaccharides. This process is highly operable, has simple steps and is environmentally friendly, providing strong support for research on the skin care mechanism of Cistanche polysaccharides.
Fan Qianwen et al. [18] invented a patent for the extraction of an oligosaccharide mixture from Cistanche tubulosa bulbs. The bulbs were defatted and dried, then extracted multiple times with boiling water. The filtrate was combined, concentrated, and then protein was removed with trichloroacetic acid. The solution was neutralized with alkali, dialyzed with flowing water for 3 days, and then the internal liquid was concentrated and centrifuged to remove the precipitate, the supernatant was precipitated with an organic solvent, the precipitate was collected by centrifugation and successively washed with anhydrous ethanol and acetone, and then vacuum-dried to obtain a dried Cistanche tubulosa bulb polysaccharide extracted by water. The resulting polysaccharide was degraded with strong acid and high temperature to remove acidic substances, and the supernatant was collected after centrifugation to prepare a Cistanche extract containing a mixture of oligosaccharides. This extract has the effect of delaying skin aging in cosmetics.
Huang Xiang et al. [19] extracted Cistanche deserticola, and the content of five phenylethanoid glycosides, namely echinacoside, acteoside, cistanoside A, isacteoside, and 2'-acetylacteoside, was as the evaluation criteria, and HPLC was used for detection and analysis. The influence of each parameter on the extraction process was optimized using single-factor experiments and response surface optimization design, and the optimal extraction process was established: extraction with 55.14% methanol, solid-liquid ratio: 1:46.39, extraction for 38.5 min, and drying at 80 °C.
3 Whitening and anti-aging mechanism of action
Skin ageing is a complex biological and cellular process involving changes in the ultrastructure and biochemistry of the skin. Lifestyle factors (such as habits, diet and smoking), diseases (such as diabetes) and gravity all have an effect on skin ageing. During the ageing process, both endogenous and exogenous ageing factors come into play. Endogenous aging is the natural, gradual aging process within the body. To some extent, endogenous aging is a programmed aging process that is regulated by the cell's internal biological clock, which determines the number of cell replications the cell undergoes during the aging process and the period required for each replication. Exogenous factors also cause skin aging. Exogenous aging mainly refers to the influence of environmental factors, such as reactive oxygen species (ROS), UV radiation, and free radicals, which have a cumulative effect on the skin and lead to the degradation of collagen and the cell matrix. UV radiation causes photoaging to deepen the endogenous aging process. Various external factors cause the collagen and elastin proteins in skin tissue to denature, reducing skin elasticity, causing wrinkles to form, and promoting skin aging.
3.1 Anti-aging
The mechanism of action of skin anti-aging is mainly to protect the skin's internal cells from damage caused by the external environment and self-aging, enhance the activity of immune cells, improve antioxidant effects, and repair the skin barrier, thereby preventing skin aging and maintaining a healthy skin condition. The anti-aging effect of Cistanche deserticola is mainly caused by D-mannitol, polysaccharides (CDPS), and phenylethanoid glycosides, which can significantly delay skin aging and increase the elasticity of the skin by increasing the content of collagen fibers [20,21]. Data show that 1% D-mannitol and 1% Cistanche polysaccharide can both increase the intracellular hydroxyproline content of animal skin cells, increase the content of collagen fibers, improve skin elasticity, and thus delay skin aging [21]. In addition, RT-PCR experiments showed that the expression of collagenase and elastase mRNA in melanoma cells treated with 4 mg/mL Cistanche deserticola extract was sharply reduced, which in turn caused the activity of collagenase, elastase and hyaluronidase to decrease, thereby slowing down skin aging [22].
Liu Yuanyuan et al. [23] studied the effect of Cistanche deserticola polysaccharides on skin aging. 20–50 mg/mL Cistanche deserticola polysaccharides can protect HaCaT cells from UVB irradiation damage, inhibit the continuous reaction of oxygen free radicals, and improve the self-healing ability of skin tissue. In particular, skin cells after UVB irradiation will be protected from UVB damage, thereby delaying skin aging. Studies have shown that CDPs have a proliferative effect on macrophages, lymphocytes and fibroblasts [24-27], and can promote cell proliferation and differentiation of megakaryocytes and erythroid progenitor cells [28]. Its polysaccharide extract promotes the proliferation of splenocytes, activates dendritic cells and the immune response [29]. Cistanche water extract can induce a strong and long-lasting antigen-specific immune response by activating dendritic cells. Dendritic cells participate in the regulation of the expression of maturation markers and cytokines through the TLR4-related NF-κB pathway. Studies have found that the active ingredient in Cistanche deserticola, phenylethanoid glycosides, also has a certain delaying effect on skin aging. Cistanche deserticola phenylethanoid glycoside emulsion can effectively absorb ultraviolet radiation from sunlight [30], and can better prevent and alleviate the damage to the skin and the photoaging effect of ultraviolet radiation [31]. In addition, the phenylethanoid glycoside extract from Cistanche has a strengthening effect on superoxide dismutase (SOD) [32], which can remove free radicals in the body, achieving an anti-oxidant and anti-aging effect.
3.2 Whitening
The most important determinant of skin tone is melanin, which is produced by melanocytes in the basal layer of the epidermis under stress conditions. The biosynthesis of skin melanin is a hydroxylation reaction of tyrosine in the body, catalyzed by the key enzyme tyrosinase to synthesize L-dopa, which is then oxidized to form dopaquinone. Dopaquinone undergoes a series of metabolic transformations, rearrangements, and combinations, and finally binds to proteins to form melanin, causing the skin to tan.
Therefore, inhibiting the activity of tyrosinase and targeting tyrosinase is the main direction of research and development of whitening cosmetics [33-36]. Compounds with potential whitening effects are mainly those containing benzene rings or phenolic hydroxyl groups, such as the commonly used whitening agents arbutin and kojic acid. According to literature reports, more than 125 different genes regulate melanin pigmentation. Some important biological functions of melanocytes, such as cell differentiation and pigmentation or melanin production, are related to these genes.
An important member of the melanocyte development transcription factor is the microphthamia transcription factor (MITF), which initiates the transcription process by activating the expression of the TYR, TRP-1, and TYR-2 genes [37]. Phenylethanoid glycosides are a class of natural glycosides formed by an ester bond and a glycosidic bond connecting the parent ribose group, phenylacrylic acid, and part of phenylethanol. Yang Jianhua et al. [38] found that 1.5–3.0 mg/mL phenylethanol glycosides can inhibit tyrosinase activity. The effect is a competitive reversible inhibition, and the whitening effect is better than arbutin. It whitens by inhibiting the production of melanin in human epidermal melanocytes.
Wu Shanshan's research report [39] found that 400 μg/mL Cistanche tubulosa phenylethanoid glycosides can effectively inhibit the tyrosinase inhibitory activity in a human skin melanocyte model caused by UVB radiation, promote apoptosis of human skin melanocytes, reduce the content of melanin granules, significantly down-regulate the mRNA transcription levels of melanin synthesis-related genes TYR, TPR-1 and MITF, and reduce the protein expression of TYR, TRP-1, TRP-2 and MITF. These results all indicate that Cistanche phenylethanoid glycosides can significantly inhibit tyrosinase activity, laying a theoretical foundation for the development of Cistanche phenylethanoid glycosides in the field of natural whitening cosmetics. Cistanche total glycosides have the effect of inhibiting skin pigmentation and whitening, and can be used to prevent and treat pigmentation-related skin diseases, or used in the development of cosmetics for whitening and freckle removal.
4 Conclusion
As the external environment continues to deteriorate, the demand for whitening and anti-aging cosmetics is also increasing due to changes in skin tone such as freckles, birthmarks, and sunspots. However, most of the efficacy studies on Cistanche extract have been carried out in vitro, using mouse skin models and nematode models to infer the mechanism of action of whitening and anti-aging. There is a lack of actual human skin care efficacy studies. Therefore, the whitening and anti-aging skin care efficacy of Cistanche extract the whitening and anti-aging skin care effects still need to be tested using human efficacy data, so that the effects of Cistanche extract in skin repair, delaying skin aging, and whitening can be further understood, so that Cistanche-based cosmetics can be sold well in the market.
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