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. 2017 Jul 25;7:6413. doi: 10.1038/s41598-017-06844-2

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© The Author(s) 2017

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Hyperoside pre-treatment reduced the increased heparanase expression induced by ROS in cultured podocytes. (A) The dose-response effect of xanthine/xanthine oxidase (X/XO) on podocytes heparanase mRNA expression. Podocytes were incubated with 0, 3, 10, 30, 100 µM xanthine plus equal doses of xanthine oxidase (mU) for 18 h. (B) The dose-response effect of NADH on podocyte heparanase mRNA expression. Cultured podocytes were incubated with 0, 0.3, 1, 3, 10 mM NADH for 18 h. (C,D) Podocyte heparanase mRNA expression after incubation with X/XO (30 µM/mU) or NADH (3 mM) for 0, 6, 12, 18 and 24 h. (E) Hyperoside pre-incubation attenuated increased heparanase mRNA expression induced by extracellular ROS. Control: vehicle incubation for 18 h; X/XO: 30 µM xanthine plus 30 mU xanthine oxidase incubation for 18 h; LHPS or HHPS: 30 or 100 μg/ml hyperoside pre-incubation for 1 h and followed by X/XO (30 µM/mU) exposure. (F) Hyperoside pre-incubation attenuated increased heparanase mRNA expression induced by intracellular ROS. Control: vehicle incubation for 18 h; NADH: 3 mM NADH incubation for 18 h; LHPS or HHPS: 30 or 100 μg/ml hyperoside pre-incubation for 1 h and followed by NADH exposure. (G,H) Hyperoside pre-incubation attenuated heparanase protein expression induced by X/XO by western blot analysis. The columns showed the amount of heparanse protein levels relative to β-actin. Control: PBS incubation for 24 h; X/XO: X/XO (30 µM/mU) incubation for 24 h; LHPS or HHPS: 30 or 100 μg/ml hyperoside pre-incubation for 1 h and followed by X/XO (30 µM/mU) exposure. Data are presented as mean ± SD. P < 0.05 is statistically significant. ^aIndicates significant vs. Control, ^bindicates significant vs. ROS.