| Objective: The purpose of this study was to observe the effect of Codonopsis pilosulapolysaccharide on oxidation damage and gene expression profile of skin tissue in aged miceinduced by D-galactose, and to explore whether Codonopsis pilosula polysaccharide candelay skin aging or not and its cell and molecular biology mechanism, and further, to discussthe inherent relationship between the Traditional Chinese Medicine theory of tonifying Qiagainst aging and the free radical theory.Methods: Two-month old male Kunming mice(120)were randomized into blank controlgroup, aging model group, Codonopsis pilosula polysaccharides low dose group, Codonopsispilosula polysaccharide high dose group and vitamin E group. Aging models were made byD-galactose, and drugs intervention started on the eleventh day of modeling. After32days oftreatment, sample materials were obtained for detection, including the skin general morpho-logical observation, the comparison of skin tissue SOD activity, ATP enzyme activity, MDAcontent, LF content and HYP content, to observe the effects of Codonopsis pilosula polysac-charide on the pathological structure and antioxidative ability of mice skin. Whole genomegene chip technology was applied to screen genes with differential expression in mouse skintissue by Codonopsis pilosula polysaccharide. GO database, NCBI database and KEGGdatabase were applied to analyze the function and signal pathway of significant expressiongenes. RT-PCR technology was used to validate the results of gene chip.Results:(1) The results of skin histopathology: compared with the blank control group, skintissue of the aging model group was thinner, with less skin appendages, less collagen fibers,loose arrangement; compared with the aging model group, the Codonopsis pilosulapolysaccharide high dose group got thicker skin, more collagen fiber, arranged more neatly.Under the electron microscope, skin tissue of the aging model mice got more nuclearheterochromatin, and with scarce cytoplasm; more dilatation rough endoplasmic reticulum,more lipofuscin granules in the cytoplasm, and less collagen fiber; skin tissue cells of the highdose Codonopsis pilosula group and the vitamin E group had no dilatation rough endoplasmicreticulum, less lipofuscin granules in the cytoplasm, more collagen fiber; skin tissue cells of the low dose Codonopsis group got slightly dilated endoplasmic reticulum and more collagenfiber.(2) Effects of Codonopsis pilosula polysaccharide on antioxidant ability of mice skin:compared with the blank control group, the activity of SOD in the aging model groupdecreased significantly (P<0.05), ATP enzyme activity decreased significantly (P<0.05); thecontent of MDA and LF increased significantly (P<0.05); the content of HYP decreasedsignificantly (P<0.05). Compared with the aging model group, SOD and ATP enzyme activityof the Codonopsis pilosula polysaccharide high dose group increased significantly (P<0.05),the content of MDA and LF decreased significantly (P<0.05), HYP signifycantly increased(P<0.05). Compared with the vitamin E group, ATP enzyme activity, SOD enzyme activity,MDA and HYP content of the Codonopsis pilosula polysaccharide high dose group had nosignificant difference(P>0.05), the Codonopsis pilosula polysaccharide low dose group wasbetween the vitamin E group and the aging model group.(3) Effects of Codonopsis pilosula polysaccharide on the gene expression of skin agingmodel mice: as the results showed, after Codonopsis pilosula polysaccharide intervention,118genes got significant differential expression, which85up-regulated and33down-regulated,mainly involved the functions of metabolism, apoptosis, immune response, oxidative stress,DNA damage, protein modification, gene transcription and skin morphology etc. KEGGdatabase was used to analyze significantly changed signal pathway because of Codonopsispilosula polysaccharide, combined with the research background of skin aging,6significantlychanged signal pathways were screened, which5up-regulated and1down-regulated.(4) The RT-PCR experimental results proved that the verified genes and the gene chipwere concerted, with the same upward or downward trend, indicating that the result of genechip was reliable.Conclusion:(1) The results show that Codonopsis pilosula polysaccharide can improve the patho-logical changes of the D-galactose induced aging mice skin tissue, enhance the antioxidantcapacity and collagen fibers content of the skin tissue of aging mice, which is helpful to delaythe aging process.(2) Codonopsis pilosula polysaccharide can delay skin aging process through the regulation of multiple genes and signal pathways, revealling that Codonopsis pilosulapolysaccharide can delay the senility of the skin by through multiple targets and pathways.(3) Nourishing Ying and Wei can delay the skin aging of the mice, and there is a closerelationship between the aging theory of Traditional Chinese Medicine Ying Qi and Wei Qideficiency and oxidative free radical theory of aging. |
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