Korean J Physiol Pharmacol.  2023 Nov;27(6):513-520. 10.4196/kjpp.2023.27.6.513.

Cornuside inhibits glucose-induced proliferation and inflammatory response of mesangial cells

• Li X
• Guo L
• Huang F
• Xu W
• Peng G

• KMID: 2547317
• DOI: http://doi.org/10.4196/kjpp.2023.27.6.513

Abstract

Cornuside is a secoiridoid glucoside compound extracted from the fruits of Cornus officinalis. Cornuside has immunomodulatory and anti-inflammatory properties; however, its potential therapeutic effects on diabetic nephropathy (DN) have not been completely explored. In this study, we established an in vitro model of DN through treating mesangial cells (MMCs) with glucose. MMCs were then treated with different concentrations of cornuside (0, 5, 10, and 30 μM). Cell viability was determined using cell counting kit-8 and 5-ethynyl-2′-deoxyuridine assays. Levels of proinflammatory cytokines, including interleukin (IL)-6, tumor necrosis factor-α, and IL-1β were examined using enzyme-linked immunosorbent assay. Reverse transcription quantitative real-time polymerase chain reaction and Western blotting were performed to detect the expression of AKT and nuclear factor-kappa B (NF-κB)-associated genes. We found that cornuside treatment significantly reduced glucose-induced increase in MMC viability and expression of pro-inflammatory cytokines. Moreover, cornuside inhibited glucose-induced phosphorylation of AKT and NF-κB inhibitor alpha, decreased the expression of proliferating cell nuclear antigen and cyclin D1, and increased the expression of p21. Our study indicates that the anti-inflammatory properties of cornuside in DN are due to AKT and NF-κB inactivation in MMCs.

Keyword

Complementary therapies; Diabetic nephropathies; Glucose; Growth; Inflammation

Figure

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  Fig. 1 Glucose treatment increased viability of MMCs. (A) The chemical structure of cornuside. (B) CCK-8 assay detected the cell viability after different doses of glucose treatment (0, 10, 25, 50, and 100 mM). Values are presented as mean ± SD. MMCs, mesangial cells; CCK-8, cell counting kit-8; OD, optical density. *p < 0.05, **p < 0.01.

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  Fig. 2 Cornuside reduced high glucose-induced over-proliferation of MMCs. CCK-8 (A) and EdU (B) assays assessed the viability and proliferation in high glucose-induced MMCs after different doses of cornuside treatment (0, 5, 10, and 30 μM). Values are presented as mean ± SD. MMCs, mesangial cells; CCK-8, cell counting kit-8; EdU, 5-ethynyl-2′-deoxyuridine; OD, optical density. *p < 0.05, **p < 0.01, ***p < 0.001.

• 

  Fig. 3 Cornuside inhibited the production of IL-6, TNF-α and IL-1β in high glucose-treated MMCs. ELISA was conducted to determine the secretions of IL-6, TNF-α and IL-1β in high glucose-induced MMCs after different doses of cornuside treatment (0, 5, 10, and 30 μM). Values are presented as mean ± SD. MMCs, mesangial cells; IL, interleukin; TNF-α, tumor necrosis factor-α. *p < 0.05, **p < 0.01, ***p < 0.001.

• 

  Fig. 4 Cornuside inhibited the AKT and NF-κB signaling pathways in high glucose-treated MMCs. (A) RT-qPCR analysis measured the mRNA levels of PCNA, cyclin D1, p21 and AKT in high glucose-treated MMCs after different doses of cornuside treatment (0, 5, 10, and 30 μM). (B) Western blot analysis measured the protein levels of p65, PCNA, cyclin D1, p21, AKT, p-AKT, p-IκB-α, COL1 and fibronectin in high glucose-induced MMCs after different doses of cornuside treatment (0, 5, 10, and 30 μM). (C) Western blot analysis measured the protein levels of p65 (nuclear) and p65 (cytosol) in high glucose-treated MMCs after different doses of cornuside treatment (0, 5, 10, and 30 μM). Values are presented as mean ± SD. MMCs, mesangial cells; NF-κB, nuclear factor-kappa B; RT-qPCR, reverse transcription quantitative real-time polymerase chain reaction; PCNA, proliferating cell nuclear antigen. *p < 0.05, **p < 0.01, ***p < 0.001.

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