Hair is a unique feature of mammals, covering almost the entire body surface, which undertakes a variety of physiological functions, including avoiding or reducing the body from various damages (such as ultraviolet rays, trauma) and regulating body temperature [38]. It has not only a cosmetic effect, but also a sign of body health [38]. Although modern medical methods can effectively improve hair loss in a short period of time, the treatment options available so far are extremely limited [39]. The Chinese nationality has a long history, and has many knowledges for preventing and curing hair loss, which have been passed on through generations. Based on these knowledges, many safe and effective ingredients were discovered and developed with scientific theories and methods, which are benefit for hair growth. O. vulgatum, is a kind of fern, which has been used as food and dietary supplement in Yunnan, Guizhou and Sichuan Provinces in China since ancient times [40]. However, there are few studies on the chemical consists and biological activity of O. vulgatum in decades. Hence, we did some research to demonstrate and evaluate cell growth promoting activity of it.
In this study, we used reflux extraction and fractional extraction to prepare the extract of O. vulgatum L. to obtain ethyl acetate fraction (OpvE). UPLC-Q/TOF-MS was engaged in identifying components in OpvE. 3-O-methyl-quercetin-7-O-diglucose-4'-O-glucoside, linoleic acid, quercetin, Ophioglossum vulgatum alcohol, etc. 24 ingredients were identity to TCM database of the SCIEXOS. Based on the prediction of these chemical components by network pharmacology, we speculate that OpvE has the activity of promoting cell proliferation and energy metabolism. We verified the analytic results with some studies, in vitro. In HaCaT cell assays, OpvE (0.98 to 250 µg/mL) showed promoting cell proliferation activity. And the results indicated that OpvE could increase the intracellular ATP content and CAT activity at a relative low concentration, which were 7.5 and 30 µg/mL. At the same time, the transcription of the skeletal proteins COL1A1 and FN1, and the cell signaling factor TNF-α are also stimulated with OpvE treatment (15–60 µg/mL).
ATP (adenosine triphosphate) is one of mainly energy-carrying molecule in all living creatures, signaling molecular, as well as basic material of DNA synthesis [41]. Therefore, increased ATP concentration in cell is one of the key signal for cell multiplication. Mitochondria are the organelle where oxidative phosphorylation and ATP synthesis takes place, a process of oxygen free radicals generation [42]. It has been known that free radicals can directly impair function and structure of micro mitochondria. This is a key factor for inducing and accelerating diseases and aging [43]. In particular, increasing the anti-oxidation capability of cell is specially important for fast growing cell [44]. CAT in mitochondria is an essential antioxidant enzyme [45]. According to the results of the current research, OpvE promoted cell growth through improve ATP levels and enhance the activity of CAT enzymes in mitochondria. This can effectively prevent fast growing induced cell damage.
The hair follicle regeneration process is still less known. The previous studies reported that increasing the content of type I collagen in skin cells can not only alleviate skin aging, but also benefit the regeneration of hair follicle stem cells [46]. Collagen type I alpha, COL1A1, is a component of extracellular matrix and is closely related to cell growth, proliferation and differentiation [47]. Fibronectin 1 (FN1) is a kind of fibronectin, which is mainly distributed in connective tissue and the basement membrane [48]. It has the physiological function of promoting the connection between cells and fibrous matrix, and participates in cell adhesion, differentiation, wound healing, blood coagulation and body defense [48].Our research results show that the extract of O. vulgatum promotes the transcription of structural proteins COL1A1 and FN1 and cell proliferation regulator (TNF-α) in cells. The results indicated the potential hair follicle promotional function of OpvE.
Ferns are an important evolutionary marker for land plants on the earth, which occupy an important position in the composition of the flora. There are a lot of medicinal value ferns used by Chinese since ancient time. Such as Lycopodium annotinum L., Selaginella tamariscina, Equisetum hyemale L., Lygodium japonicum (Thunb.) Sw., Davallia mariesii Moore ex Bak., Scutellaria barbata D. Don, Pyrrosia lingua (Thunb.) Farwell, Dryopteris crassirhizoma Nakai (Rhizoma Dryopteris Crassirhizomae) are listed in the Pharmacopoeia of the People's Republic of China [49]. In comparison of aforementioned ones, O. vulgatum is not a popular used herb. It was reported high content of unsaturated fatty acids [50, 51], as well as galactoglycerolipids [52], flavonoids [53, 54] and polysaccharides [55] in O. vulgatum. Even though, the pharmacochemical characterization was still rarely known. This limited the bioactivity studies.