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Published:
Mar 6, 2018DOI:
10.4314/tjpr.v17i2.5Keywords:
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Main Article Content
Glechoma longituba (Lamiaceae) alleviates apoptosis in calcium oxalate-induced oxidative stress in kidney proximal tubule epithelial cell line, HK-2
Abstract
Purpose: To investigate the effects of Glechoma longituba on calcium oxalate (CaOx)-induced stress in HK-2 cells as a possible treatment strategy for nephrolithiasis (kidney stones).
Methods: Human kidney HK-2 cells were treated with CaOx and Glechoma longituba at different concentrations. The levels of reactive oxygen species (ROS), lactate dehydrogenase (LDH), and malondialdehyde (MDA) were measured. Cell apoptosis and viability were assessed by flow cytometry and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays, respectively, while apoptosis-related proteins were determined using western blotting. The levels of the nuclear factorerythroid 2-related factor 2 (Nrf2), heme oxygenase-1 (HO-1), and NADPH-quinone-oxidoreductase 1 (NQO-1) genes were evaluated using quantitative real-time polymerase chain reaction (qRT-PCR). Using lentivirus, Nrf2 was knocked down in HK-2 cells, and this was confirmed by both qRT-PCR and western blotting. Scramble and si-Nrf2 transfected HK-2 cells were treated with CaOx and Glechoma longituba, and ROS levels and apoptosis were also assessed.
Results: CaOx significantly increased the levels of ROS, LDH and MDA, while Glechoma longituba pretreatment attenuated these elevations in a dose-dependent manner. CaOx treatment increased cell apoptosis and decreased cell viability (p < 0.05), while Glechoma longituba pre-treatment abolished these effects in a dose-dependent manner. Glechoma longituba pre-treatment significantly upregulated the expressions of Nrf2, HO-1 and NQO-1 (p < 0.05). In HK-2 cells, Si-Nrf-2 attenuated the effects of Glechoma longituba pre-treatment on cell oxidative stress and apoptosis induced by CaOx.
Conclusion: Glechoma longituba pre-treatment attenuates cell apoptosis and oxidative stress induced by CaOx via Nrf2/HO-1 signalling pathway. Thus, the plant is a potential source of agents for the treatment of nephrolithiasis.
Keywords: Glechoma longituba, Nephrolithiasis, Nuclear factor-erythroid 2-related factor 2, Oxidative stress, Apoptosis
