Presentation Information
[2Cp02]Effect of apple pomace extract on human skin fibroblasts
*Naoki Nanashima1, Hayato Maeda2, Akira Nakajima2, Makoto Nishizuka2, Tsuyoshi Narumi3, junji ichita3,4, Koh Itoku3 (1. Aomori Univ. of Health & Welfare, 2. Hirosaki Univ., 3. NIHON HARUMA Co.LTD, 4. Shibata Gakuen Univ.)
Keywords:
apple pomace,skin fibroblasts
Objective: Human skin fibroblasts (HSFs) are cells in the dermis that produce collagen and hyaluronic acid, which are related to skin appearance. Apple pomace is generated during the production of apple juice; however, most of this apple pomace is discarded. Apple pomace contains beneficially affect human health; however, no studies have been conducted on the effects of apple pomace on the skin. As such, we analyzed the effects of apple pomace extract (APE) on HSFs.
Methods: APE was added to the culture medium for HSF, and after 72 hours of culture, Cell Counting Kit-8 was added to evaluate cell proliferation. RNA was extracted 24 hours after the APE was added, and the gene expression levels were determined using next-generation sequencing transcriptome and Gene Ontology (GO) analyses as well as real-time PCR. The amounts of type I collagen and hyaluronan were quantified using an ELISA. APE was added to the HSFs, and the phosphorylation of p38MAPK, ERK, and AKT was analyzed using Western blotting 30 min later.
Results: The HSF proliferation increased at APE concentrations of 10 and 20μg/mL compared with the control. The application of APE increased the expression of hyaluronan synthases HAS1, HAS2, and HAS3 and decreased the expression of the hyaluronan decomposition enzyme HYAL1 in HSFs. In addition, the type I collagen produced by the HSFs was higher at APE concentrations of 5.0 and 20μg/mL compared with those in the control. Hyaluronan levels were higher at APE concentrations of 1.0, 5.0, and 20μg/mL. Adding APE to the culture induced the phosphorylation of p38MAPK, ERK, and AKT in the HSFs. Most of the top ten terms in the GO analysis were related to mitosis, such as nuclear chromosome segregation and mitotic nuclear division.
Methods: APE was added to the culture medium for HSF, and after 72 hours of culture, Cell Counting Kit-8 was added to evaluate cell proliferation. RNA was extracted 24 hours after the APE was added, and the gene expression levels were determined using next-generation sequencing transcriptome and Gene Ontology (GO) analyses as well as real-time PCR. The amounts of type I collagen and hyaluronan were quantified using an ELISA. APE was added to the HSFs, and the phosphorylation of p38MAPK, ERK, and AKT was analyzed using Western blotting 30 min later.
Results: The HSF proliferation increased at APE concentrations of 10 and 20μg/mL compared with the control. The application of APE increased the expression of hyaluronan synthases HAS1, HAS2, and HAS3 and decreased the expression of the hyaluronan decomposition enzyme HYAL1 in HSFs. In addition, the type I collagen produced by the HSFs was higher at APE concentrations of 5.0 and 20μg/mL compared with those in the control. Hyaluronan levels were higher at APE concentrations of 1.0, 5.0, and 20μg/mL. Adding APE to the culture induced the phosphorylation of p38MAPK, ERK, and AKT in the HSFs. Most of the top ten terms in the GO analysis were related to mitosis, such as nuclear chromosome segregation and mitotic nuclear division.
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