| Objective:1. To study the determination method of total flavones in Oxytropis. falcata Bunge (TFOF).2. To study a technic process, this could be used to enrich the total flavones from O. falcata Bunge (TFOF) with macroporous adsorptive resins.3. To study the oil in water (O/W) cream matrixes with different compositions, and prepare the one percent O. falcata total flavones cream (1% TFC), then the stabilities of the cream will be evaluated.4. To research the protective effects of 1% TFC on the destructed skin induced by moderate-wave ultraviolet (UVB) irradiation, to study the ability of anti-UVB by rats'appearance and changes of pathological, as well as some changes of biochemical indicators in the tissue homogenate of rat skin.Methods:1. Rutin was used as reference substance and the different of coloring reagent was chosed to determine the content of the total flavones of O. falcata Bunge. Before extracting and enriching the total flavones from O. falcata Bunge, the adsorbability and availability of macroporous adsorptive resins were investigated by determining the content of flavonoid glycoside using colorimetric method.2. Different cream matrixes with diverse composition of oil in water (O/W) were chosen and the one percent O. falcata total flavones cream (1% TFC) was prepared using mixing method. The quality and stability of the creams were evaluated by observating the extrinsic and microcosmic and determinating the contents of total flavones.3. The dorsal skin of rats was depilated by 8% Na2S and the changes would recover one week. The depilated rats were irradiated with UVB, acute UVB damage mice model was set up by adjusting the irradiate time. 4. The depilated rats were divided into four groups:normal control, UVB control, UVB+GYC group and UVB+1% TFC group. The UVB+GYC and UVB+1% TFC groups were treated with 10 mg/cm2 drugs after anesthetized. The UVB control was irradiated, but not the normal control. The weight of rats was recorded and their skin was observed before irradiated daily. After seven days, the rats were killed within 24 hours. Pathological sections of rat's skin were made and observed. The rat's irradiated skin was cut (0.3 g for each) and saved in -20℃. The biochemical indicators of the skin including:superoxide dismutase (SOD), malondialdehyde (MDA), glutathione peroxidease (GSH-Px), glutathion-s-transferase (GST), glutathione (GSH), catalase (CAT), hydroxyproline (Hyp) and hydroxy radical (-OH) were diagnosed with ultraviolet spectrophotometry.Results:1. Aqueous extract of O. falcata Bunge was hydrolyzed by 2.5 M hydrochloric acid methanol and colorated by 3.0% zirconium oxychloride (ZrOCl2). The content of total flavones in O. falcata Bunge was determined as 5.8%. Meanwhile, the content of total flavonoids in total flavonoids extract was 26%, it was about 5 times comparing with the total flavonoids content before enriched.2. Stearic acid 3 g, white vaseline 3 g, satiric-glycerol ester 1 g, span-60 0.8 g, and liquid paraffine 4.5 g were used as the oil phase, and tween-80 2.5 g, glycerol 5 g, methyl-p-hydroxybenzoate 0.1 g, and distilled water 40 ml were used as the aqueous phase to prepare the cream, this cream possessed better stability and was easy to use and to clean.3. The rats were irradiated with UVB phototherapy at a distance of 20 cm with 5 min once a day for seven days. Then the dorsal skin of rats was red, intumesce and ulcerate, touching like leather. At this time, the rats displayed typical acute UVB damage.4. The pathological section of skin of rat was made and dyed by fast H-E stain (HE). The skin texture was obsvered by microscope. The results showed that dermal ridge was clear and the tissue was no inflammatory reaction in UVB+1% TFC group and UVB+GYC group, and no changes compared with the normal control. The UVB-irradiated control showed thickened epidermis, hyperkeratosis and focal parakeratosis. Moreover, the plasmexhidrosis was observed in the corneum; acanthosis, local microabscess was observed in stratum spinosum; edema and bleeding were observed in superficial dermis, meanwhile, many infiltrated neutrophil and small amounts of diapedetic red cell were observed too.5. The oxidative indicators in rats' skin were determined. The cream could improve the activities of SOD (P<0.001), GSH-Px (P<0.001), GST (P<0.05) and CAT (P<0.01), the content of Hyp (P<0.001) was increased and contents of MDA and·OH were decreased (P<0.001) significantly. It could also decrease the activities of GSH (P<0.001) significantly. All of these indicated the cream could protect skin from ultraviolet significantly.Conclusions:1. Acid hydrolysis and 3.0% zirconium oxychloride (ZrOCl2) chromatometry could be used to determine the content of total flavones from O. falcata Bunge, and this colorimetric method was simple and easy to opreate. D-101 macroporous resin could be effective to enrich and extract the TFOF with better adsorbability and higher repetat availability.2. The TFC was glossy and stability, which was prepared by the method of prescriptional optimization.3. In vivo, the cream of the total flavonoids from O. falcata Bunge could protect the destructed skin of rat and prevent the damage in appearance and pathalogy from UVB irradiation. Meanwhile, it could also prevent changes of oxidative indicators in rat skin. |
PDF Full Text Request