| ObjectiveSenescent is known to be a naturally complicated multifactorial process, which associated with a gradual slow decline in the physiologic function of organisms owing to the growth of age and stimulation of other factors. It is characterized with wrinkles, progressive memory loss, and all the prominent organs (brain, liver, kidney, heart and lung) dysfunction and so on. Although the aging process is the inevitable fate of all organisms, it is can be delayed by some effective methods and the inhibition of aging is the desire and pursuit among all the human beings. Nowadays, population ageing is a serious phenomenon for nearly every country in the world, along with age related diseases including dementia, alzheimer disease and so on, which brings tremendous challenges in the world. Population aging not only affects the elderly health, but also causes the medical and social issues. Therefore, the study of anti-aging has been paid close attention in the life sciences community, and how to prevent aging-related diseases is extremely urgent.Recently, herbal medicines have been attracting a great deal of attention as alternative and supplemental medicines. Chrysanthemum indicum Linnen (C. indicum), an oriental traditional medicine, has been used to treat inflammation, immune-related disorders, hypertension, several infectious diseases, with a high efficacy and minimal side effect profile. It was reported that the prominent constituents in C. indicum, including flavonoids, terpenoids, phenolic compounds, volatile oil and terpenes, could be responsible for its therapeutic properties, whereas these components are unstable and can be easily decomposed facing heat. At the same time, supercritical carbon dioxide extraction technology (SCFE) can effectively protect these heat-sensitive chemicals with low toxicity. Based on this, SCFE extract of C. indicum (CISCFE) is used in this study. Besides, modern pharmacological studies have shown that C.indicum exerted obvious anti-oxidation activities and the previous study in our lab have reported that it had remarkable anti-inflammation properties, but its anti-aging effect to our knowledge has been scarcely studied. Moreover, aging is considered to be closely connected with oxidation and chronic inflammatory states, and C. indicum had anti-oxidation and anti-inflammation effects, thus suggesting it may be a promising anti-aging drug.Therefore, the present study was carried out to investigate the effect of CISCFE in ameliorating D-galactose-induced senescent and UV-induced photoaging and elucidated the possible mechanisms.Methods1. The anti-aging effect of CISCFEFemale Kunming mice were subcutaneously injected D-galactose for 10 weeks to establish the D-Galactose-induced senescent model, with oral administration of CISCFE (100,150 and 300 mg/kg). After 8 weeks of treatment, the animal general features were observed including food and water intake, weight gain, mental state, fur color, behavior and urine. Besides, the thymus index and spleen index were measured. Moreover, morphological alterations in hippocampus and liver tissues were checked using H & E staining method, to study the anti-aging effect of CISCFE.2. The mechanism investigation of the anti-aging effect of CISCFEMice were administered CISCFE simultaneously to inhibit senescent. The content of T-AOC, the activities of antioxidant enzymes (SOD, CAT, GSH-Px) and the level of MDA in brain and liver tissues were evaluated by biochemical quantitative methods. Meanwhile, the concentrations of AST,ALT, TNF-a,IL-1 β and IL-6 in blood serum were analyzed via enzymatic technology. The expressions of Bax, Bcl-2, Caspase-3 were detected by Western-blot.3. The anti-photoaging effect of CISCFEMice were repeatedly exposed of UVA and UVB irradiation to establish skin photoaging model via dose gradually minimal erythema dose (MED) method, and CISCFE (1.2,3.6 and 12.0 mg/mouse) were applied topically to the shaved dorsal skin. Skin macroscopic visual score, the time of pinch test, as well as, skin-fold thickness were estimated, therefore, to study the anti-photoaging effect of CISCFE.4. The mechanism investigation of the anti-photoaging effect of CISCFEMice were topically administered CISCFE to attenuate skin photoaging. Activities of SOD, CAT, GSH-Px, as well as levels of MDA in skin were determined according to assay kits. ELISA analysis was used to detect IL-1 β, IL-6, IL-10, TNF-a, MMP-1 and MMP-3 expressions. Besides, the degree of skin structure alteration and elastosis were assessed visually using H&E staining and Gomori’s aldehyde fuchsin staining. Finally, we aimed to explore the mechanisms of the anti-aging effect of CISCFE by detecting these indicators.Results1. The anti-aging effect of CISCFEResults indicated that CISCFE (especially 150 and 300 mg/kg) could improve appetite, mental condition, body weight, and fur condition of D-galactose-induced senescent model mice, increase weight gain, and inhibit the decline of thymus and spleen. Importantly, CISCFE significantly relieve the pathological changes in liver and brain tissue induced by D-galactose, via protecting the morphology and structure of neuronal cells and hepatocytes2. The mechanism investigation of the anti-aging effect of CISCFEBiochemical examination revealed that CISCFE markedly enhanced the level-of T-AOC, the activities of antioxidant enzymes (SOD、CAT、GSH-Px), lowered the content of MDA in liver and brain tissue. Besides, CISCFE inhibited the release of inflammatory cytokines (TNF-α、IL-1β、IL-6), decreased the content of AST and ALT in blood serum, down-regulated the expressions of Bax and Caspase-3 and up-regulated the expression of Bcl-2.3. The anti-photoaging effect of CISCFECompared with UV-induced photoaging model mice, CISCFE could obviously suppressed wrinkles, laxity as well as erythema. At the same time, CISCFE also decreased the time of pinch test and skin fold thickness after 10 weeks.4. The mechanism investigation of the anti-photoaging effect of CISCFEResults indicated that CISCFE could effectively ameliorate the structure and morphology of photaged-skin collagen and elastic fibers. Moreover, CISCFE also enhanced the levels of SOD, CAT, and GSH-Px, decreased the content of MDA, increase the concentrate of Hyp, reduced the secretion of inflammatory cytokines (IL-1β, IL-6, IL-10 and TNF-α), and inhibited the expression of MMP-1 and MMP-3.ConclusionSummarizing, this is the first time that the antiaging effects of CISCFE have been established in an animal model induced by excessive D-galactose and the inhibition was ascribed to the increase of antioxidant capacity and immune function, inhibition of inflammatory mediators, revealing that CISCFE was potential agent for anti-aging. Equally, anti-photo-aging effects of CISCFE have been studied using an animal model induced by UV radiation and the inhibition was ascribed to elevated collagen production via the down-regulation of MMPs, which may be mediated by the inhibition of antioxidant enzymes activities and inflammatory protein expressions, suggesting the antiphotoaging property of CISCFE. |
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