| Codonopsis pilosula(C.pilosula),as a nourishing Chinese herbal medicine,has been reported to have the effect of?tonifying the middle energy,strengthening the spleen and lungs?.It is not only widely used in medicines,health products and foods,but also in beauty and skin care products.At present,C.pilosula is often used in the form of its extract in beauty and skin care,and its functional components are not clear.Based on our team's in-depth research on C.pilosula extract in the early stage,it was found that the crude C.pilosula oligosaccharides by water extraction and ethanol precipitation shown a stronger DPPH free radical scavenging ability.The purified C.pilosula oligosaccharide(CPO)was an inulin-type fructan with a molecular weight of318Da,it shows strong antioxidant activity.This thesis firstly explains the inhibitory effect of CPO on D-Galactose(D-Gal)-induced rat senescence from multiple levels of overall animals,organs,tissues,and protein expression;secondly,through formula screening,process optimization,and efficacy evaluation,a moisturizer containing CPO was developed.The specific implementation is as follows:(1)Research on anti-aging effects and mechanism of CPOTo investigate the effect of CPO on the aging of SD rats induced by D-Gal,72SD rats were randomly assigned to six groups.The model group,three CPO groups and positive control(VE)group were injected intraperitoneally with D-Gal(200mg/kg/d)for 2 weeks.Normal group were injected intraperitoneally with the same amount of normal saline(0.9%,w/v)for 2 weeks.In addition to intraperitoneal injection from the third week,CPO(230,400 and 700 mg/kg/d),VE(200 mg/kg/d)was administrated by intragastric to three CPO groups and the positive control group for 4 weeks,respectively.While rats in the normal group and model group were orally given same amount of normal saline(0.9%,w/v)for 4 weeks,respectively.24h after the last administration,the samples such as serum and liver were collected for determination of related indicators.The results showed that CPO effectively increased the growth rate of D-Gal-induced SD rats,attenuated the decline of thymus and liver indexes,and mitigated the D-Gal-induced the pathological liver injury.CPO dose-dependently elevated the CAT,GSH-Px and SOD activities and MDA level in serum compared with model group rats;improved the activity of SOD and GSH-Px in liver and down-regulated the phosphorylation of MAPK cascade signal transduction components-ERK,JNK and p38,inhibited the activation of downstream signals NF-?B protein in hepatocytes.In addition,CPO treatment noticeably prevented the overexpression of inflammatory factors such as TNF-a,IL-6 and IL-1?in serum,and inhibited the expression levels of apoptotic proteins caspase-3,caspase-9 and the ratio of Bax/Bcl-2 in liver tissue caused by the inflammatory cascade.These results suggest that CPO significantly improved the formation of oxidative stress,inflammation and apoptosis caused by excessive ROS production induced by D-Gal,relieved liver damage and delayed aging.It could be inferred that CPO was expected to be an important function factor for anti-aging of cosmetics and other products.(2)Preparation with CPO moisturizerIn order to prepare a moisturizer with CPO as the main functional ingredient,this study determined the formulation excipients based on the six major systems of emulsified cosmetic formulations,and then combined physical and chemical indicators and fuzzy mathematical sensory evaluation methods to determine the dosage of thickener(carbomer and hydroxyethyl cellulose)and emulsifier(glyceryl stearate).The preparation process conditions(emulsification temperature,emulsification time and emulsification speed)were also optimized.The results shown that when the dosage of carbomer,hydroxyethyl cellulose and glycerin monostearate were 0.6%,1.0%and 3.0%respectively;the preparation process conditions were as follows:the emulsifying temperature was 75°C,the emulsifying speed was 120r·min-1 and the emulsifying time was 20 min,the physical and chemical properties of the moisturizer was stable and the sensory evaluation was the highest.The CPO-containing moisturizer prepared by this method had good brightness of appearance,good spreading,moistening,massiness and absorptivity of coating stage,good viscosity,brightness,smoothness,massiness,absorptivity and moisture retention after application(3)Efficacy and hygiene evaluation of CPO moisturizerSubjects applied the same amount of homemade moisturizer every day for 8weeks,and used the CBS skin analysis system to objectively evaluate the skin oil,pigment,elastin,collagen and moisture status during the experiment.At the same time,subjects subjectively assessed skin absorption,firmness,complexion and moisture status by using self-assessment questionnaires.Furthermore,the improvement effect on the skin after 56 days of using CPO moisturizer was evaluated.According to the hygiene indicators in QB/T 1857-2013 of"Moisturizer Cream",the microorganisms and heavy metals in the homemade moisturizer were tested to check whether they meet the standards.The results shown that from the measurement data of skin oils,pigments,elastin,collagen and skin dermal layer moisture,it could be seen that CPO-containing moisturizers could keep skin moisturized,improve skin dullness,and increase skin elasticity.The total number of microbial fecal coliforms,staphylococcus aureus,pseudomonas aeruginosa,molds,yeasts,and colonies were all within a limited range,and the content of heavy metals mercury,arsenic,lead,and cadmium did not exceed the standard.It was shown that the prepared CPO had the effect of effectively improving the skin condition,and the hygiene met the product requirements.Based on the research of CPO's anti-aging activity and its mechanism of action,we further developed a CPO moisturizer with moisturizing and anti-wrinkle effects.The efficacy and hygiene evaluation index results proved that the product has certain effectiveness and safety. |
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