RESUMEN
This study aimed to investigate the effect of the oral administration of sea cucumber protein (SCP) on wound healing. SCP was isolated and purified from the body wall of Stichopus japonicus. A mouse skin incision model was operated on to evaluate the wound repair effect of SCP. The histological changes in the skin at the wound sites of BALB/c mice were observed by staining with haematoxylin and eosin (H&E) and Masson's trichrome. The enzyme-linked immunosorbent assay (ELISA) was used to analyze the expression of inflammatory cytokines in BALB/c mice. The boost cell migration ability was detected by a scratch assay after HaCaT cells were cultured with digested SCP (dSCP). Western blotting and RT-PCR assays were performed to determine the mechanism of SCP promoting wound healing. As a result, the wound healing rate in the SCP high dose group was 1.3-fold, compared to that in the blank group on day 14. Also, increased epidermal thickness and 1.79-fold collagen deposition contrasted with the blank group. Additionally, SCP could up-regulate the levels of pro-inflammatory factors (IL-1ß, IL-6, TNF-α) from day 3 to 7 firstly and decreased from day 7 to 14. IL-8 expression continuously decreased while the level of anti-inflammatory factor (IL-10) increased during the healing stage. Furthermore, the cell closure area reached 67% after being treated with 50 µg mL-1 of dSCP for 48 h. Cell proliferation was associated with the dSCP-activated PI3K/AKT/mTOR pathway. Taken together, SCP can be orally used as an effective agent for wound repair.
Asunto(s)
Pepinos de Mar , Stichopus , Administración Oral , Animales , Colágeno/metabolismo , Eosina Amarillenta-(YS)/farmacología , Interleucina-10 , Interleucina-6 , Interleucina-8 , Ratones , Fosfatidilinositol 3-Quinasas/genética , Fosfatidilinositol 3-Quinasas/metabolismo , Proteínas Proto-Oncogénicas c-akt/genética , Proteínas Proto-Oncogénicas c-akt/metabolismo , Pepinos de Mar/metabolismo , Transducción de Señal , Stichopus/metabolismo , Serina-Treonina Quinasas TOR/genética , Serina-Treonina Quinasas TOR/metabolismo , Factor de Necrosis Tumoral alfa/farmacología , Cicatrización de HeridasRESUMEN
Codonopsis pilosula is a perennial species of flowering plants propagated in Northeast Asia. A water-soluble polysaccharide, CPPS(3), was extracted from the root of Codonopsis pilosula by boiled water extraction and ethanol precipitation. The molecular weight was estimated to be 7.4 x 10(4) Da determined by using Gel permeation chromatography. Monosaccharide composition and the structure of the polysaccharide were determined by gas spectroscopy, Fourier transform IR (FT-IR) spectroscopy, NMR spectroscopy and mass spectroscopy and some chemical method analysis was made. The components were galactose, arabinose and rhamnose in the molar ratio of 1.13:1.12:1. The main chain of CPPS(3) is illustrated to be (1-->3)-linked-beta-GalpNAc, (1-->3)-linked-alpha-Rhap and (1-->2,3)-beta-Galp.
Asunto(s)
Codonopsis/química , Monosacáridos/análisis , Extractos Vegetales/química , Polisacáridos/química , Cromatografía en Gel , Estructura Molecular , Raíces de Plantas , Polisacáridos/aislamiento & purificación , Solubilidad , Análisis Espectral/métodos , AguaRESUMEN
A water-soluble polysaccharide, GPB, was obtained from leaf of Ginkgo biloba L by hot water extraction followed by precipitation with ethanol and fractionation with gel chromatography. The results of HPLC with TSK-GEL column and gel filtration chromatography with Sepharose CL-6B analysis indicated GPB was uniform in polarity and its molecular weight (MW) was about 10 kDa. The structure of GPB was analysed using chemical methods, IR spectroscopy, and NMR spectroscopy. GPB has a high branched structure with polygalactose as core part of backbone. The repeating unit of polygalactose consists of 1,6-linked Galactose (Gal) and 1,3,6-linked Gal.