Effect Of Yanshou Liquid On Skin Senescence Induced By Ozone Inhalation In Rats And Its Mechanism

Objective:To obverse the effect of Yanshou decoction on aged skin in rats inhaling ozone and to explore its mechanism.Methods:Each of male SD rat was set in a stainless steel chamber, exposed to 1.2±0.2 ppm ozone for 4 hours daily in consistent 45 days. Rat from control was exposed to fresh air. Ozone concentration in the chamber was monitored by Indigo disulphonate spectrophotometry. Rats were divided into control (water), model (water), vitamin E (13.5 mg·kg-1) and Yanshou decoctions at three dosages (45,90,180 mg·kg-1). They were intragastrically administrated once daily for consecutive 45 days. The activity of serum SOD, plasma GSH-Px or plasma CAT, the level of serum LPO or dermis HYP were detected and the cutaneous histopathology was observed. Morphometry of skin epidermis thickness, collagen fiber, elastic fiber and reticular fiber ratio of changes in rats. The activity of plasma T-SOD, CuZn-SOD and Mn-SOD the level of serum LPO or dermis Hyp were detected. Immunohistochemistry, Western-blot and Picric acid-Sirius red staining which observed by polarization microscopy combined with image analysis. The differential expression proteins were analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS), and identified through searching Swiss-prot database with Mascot software.Results:Compared with normal control group, the activity of SOD, the contents of LPO in serum were decreased significantly in model group, the activity of GSH-Px and CAT in plasma were decresaed, the contents of Hyp in skin tissue homogenate were decreased significantly in model group(P<0.01) and thinning of the epidermis was present. Compared with model group, the activity of SOD, the contents of LPO in serum and the activity of GSH-Px, CAT in plasma were increased significantly in yanshou decoction low-dose, medium-dose, high-dose groups, the contents of Hyp in skin tissue homogenate were increased significantly in yanshou decoction low-dose, medium-dose groups(P<0.01 or P<0.05), a histological characteristic of chronological aging in the epidermis is a decrease of tissue thickness. However, thinning of the epidermis was present in high-dose group. The contents of soluble collagen, collagen fiber, elastic fiber and reticular fiber were increased in yanshou group decoction(P<0.01) and the epidermis was thicker in yanshou decoction group(P<0.01) compared to model group. Moreover, yanshou decoction can prevent the skin mast cells of rats from releasing granules induced by ozone. Compared with normal control group, the activity of T-SOD, Mn-SOD in plasma were decresaed, there was no apparent change in activity of CuZn-SOD in model group, the contents of LPO in serum were increased significantly in model group, collagen type Ⅰ became attenuated, they were decreased significantly(P<0.01) and the content of collagen type Ⅲ were increased significantly, they were sparsely and arranged in disorder in model group(P<0.01), the expression of TGF-β1 was decreased significantly and MMP-1 was increased significantly in model group(P<0.01). Compared with model group, the activity of T-SOD and Mn-SOD in plasma were increased significantly, while the contents of LPO in serum were decreased significantly in yanshou decoction low-dose, medium-dose, high-dose groups, the content of collagen type I and the expression of TGF-β1 were increased significantly in yanshou decoction three dose groups(P<0.01 or P<0.05), the ratio of collagen type III and the expression of MMP-1 were decreased significantly in yanshou decoction low-dose, medium-dose groups(P<0.01 or P<0.05).130 differential proteins found on the rat livers of the there groups, among them 39 in Yanshou Decoction group’ s down shift,41 in up shift. Select 62 typical differential proteins and conduct biological analysis, the results show that, after the Yanshou Decoction intervention, the typical differential protein of liver tissue expression mainly take part in the oxidation reduction reaction, Most proteins associated with the metabolic processes, proteins involved in lipid, fatty acid, coenzyme and cofactors metabolism and other metabolic processes.Conclusions:By promoting collagen synthesis, up-regulated expression of TGF-β1 and down-regulated expression of MMP-1 in yanshou decoction groups suggested that it may involve in postponing the process skin aging. Yanshou decoction can defense ability of stress, reduce inflammation, increase the overall antioxidant capacity and protect cells from apoptosis. Moreover, Yanshou decoction can reduce liver fatty acid synthesis, promote fatty acid β oxidation decomposition of energy supplied and oxidative phosphorylation to generate, ATP, so as to reduce the oxidative damage and anti-aging, which provide new clues for the research on protection mechanism of Yanshou decoction.