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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 inhibited the increased heparanase expression and promoter activity induced by high glucose exposure in cultured podocytes. (A) The dose-response effect of glucose on podocyte heparanase mRNA expression. Cultured podocytes were incubated with 5 (as the control), 10, 15, 20 mmol/L glucose or mannitol of the same osmolity with 20 mmol/L glucose for 18 h. (B) Hyperoside pre-incubation attenuated the increased heparanase mRNA expression induced by 20 mmol/L glucose. Control: PBS incubation for 18 h; Glucose: 20 mmol/L glucose incubation for 18 h; LHPS or HHPS: 30 or 100 μg/ml hyperoside pre-incubation for 1 h and followed by 20 mmol/L glucose exposure. (C,D) Hyperoside pre-incubation attenuated heparanase protein expression induced by high glucose by western blot analysis. The columns showed the amount of heparanse protein levels relative to β-actin. Control: PBS incubation for 24 h; Glucose: 20 mmol/L glucose incubation for 24 h; LHPS or HHPS: 30 or 100 μg/ml hyperoside pre-incubation for 1 h and followed by 20 mmol/L glucose exposure. (D) The dose-related effect of glucose on heparanase promoter activity. Podocytes were grown under normal glucose conditions (5 mmol/L) and transfected with HPR1 promoter–driven luciferase reporter plasmid DNA or renilla luciferase gene as an internal control. After growing in complete medium containing 5, 10, 20 or 25 mmol/L glucose for 24 h, cells were harvested and measured for luciferase reporter activity. (E) Hyperoside pre-incubation attenuated increased heparanase promoter activity. Control: PBS incubation for 24 h; Glucose: 20 mmol/L glucose incubation for 24 h; HPS + Glucose: 100 μg/ml hyperoside pre-incubation for 1 h and followed by 20 mmol/L glucose exposure. HPS: 100 μg/ml hyperoside incubation alone. The relative light unit in each sample was normalized by renilla luciferase activity. Data are presented as mean ± SD. P < 0.05 is statistically significant. ^aIndicates significant vs. Control, ^bindicates significant vs. Glucose.