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Berberine activates Nrf2 nuclear translocation and inhibits apoptosis induced by high glucose in renal tubular epithelial cells through a phosphatidylinositol 3-kinase/Akt-dependent mechanism

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Abstract

Apoptosis of tubular epithelial cells is a major feature of diabetic kidney disease, and hyperglycemia triggers the generation of free radicals and oxidant stress in tubular cells. Berberine (BBR) is identified as a potential anti-diabetic herbal medicine due to its beneficial effects on insulin sensitivity, glucose metabolism and glycolysis. In this study, the underlying mechanisms involved in the protective effects of BBR on high glucose-induced apoptosis were explored using cultured renal tubular epithelial cells (NRK-52E cells) and human kidney proximal tubular cell line (HK-2 cells). We identified the pivotal role of phosphatidylinositol 3-kinase (PI3K)/Akt in BBR cellular defense mechanisms and revealed the novel effect of BBR on nuclear factor (erythroid-derived 2)-related factor-2 (Nrf2) and heme oxygenase (HO)-1 in NRK-52E and HK-2 cells. BBR attenuated reactive oxygen species production, antioxidant defense (GSH and SOD) and oxidant-sensitive proteins (Nrf2 and HO-1), which also were blocked by LY294002 (an inhibitor of PI3K) in HG-treated NRK-52E and HK-2 cells. Furthermore, BBR improved mitochondrial function by increasing mitochondrial membrane potential. BBR-induced anti-apoptotic function was demonstrated by decreasing apoptotic proteins (cytochrome c, Bax, caspase3 and caspase9). All these findings suggest that BBR exerts the anti-apoptosis effects through activation of PI3K/Akt signal pathways and leads to activation of Nrf2 and induction of Nrf2 target genes, and consequently protecting the renal tubular epithelial cells from HG-induced apoptosis.

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Abbreviations

NRK-52E cells:

Normal rat kidney tubular epithelial cells

HK-2 cells:

Human kidney proximal tubular cell line

HG:

High glucose

MTT:

3-[4,5-dimethylthiazol-2-y]-2,5-diphenyltetrazolium bromide

DMEM:

Dulbecco’s modified Eagle medium

DMSO:

Dimethyl sulfoxide

BSA:

Bovine serum albumin

RIPA:

Radioimmune precipitation assay

TGF:

Transforming growth factor

PBS:

Phosphate-buffered saline

DCF-DA:

2,7-Dichlorofluorescein-diacetate

TBS:

Tris-buffered saline

ROS:

Reactive oxygen species

Nrf2:

NF-E2-related factor 2

ARE:

Antioxidant response elements

HO-1:

Heme oxygenase-1

STZ:

Streptozotocin streptozotocin

SOD:

Superoxide dismutase

GSH:

Glutathione

LDH:

Lactic dehydrogenase

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Acknowledgments

This work was supported by the Natural Science Foundation of China (81170561, 81170775), and Postdoctoral Science Foundation of China (2014MM551144).

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Authors and Affiliations

  1. Department of Nephrology, Liaoning Province Benxi Center Hospital, 29 Victory Road, Benxi, 117000, Liaoning, People’s Republic of China

    Xiuli Zhang

  2. Research Laboratory, Liaoning Province Benxi Center Hospital, Benxi, 117000, Liaoning, People’s Republic of China

    Xiuli Zhang, Yan Jiang, Hui He & Wei Liang

  3. Key Laboratory of Medical Cell Biology, Ministry of Education, Shenyang, 110001, Liaoning, People’s Republic of China

    Xiuli Zhang & Yue Zhao

  4. Troops of 95935 Unit, Haerbin, Heilongjiang, People’s Republic of China

    Dan Liang

  5. Department of endocrinology, The Affiliated Hospital of Mudanjiang Medical University, Mudanjiang, 157000, Heilongjiang, People’s Republic of China

    Xu Lian

  6. Department of Pathophysiology, China Medical University, Shenyang, 110001, Liaoning, People’s Republic of China

    Zhi-Hong Chi

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  1. Xiuli Zhang
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Correspondence to Xiuli Zhang.

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Zhang, X., Liang, D., Lian, X. et al. Berberine activates Nrf2 nuclear translocation and inhibits apoptosis induced by high glucose in renal tubular epithelial cells through a phosphatidylinositol 3-kinase/Akt-dependent mechanism. Apoptosis 21, 721–736 (2016). https://doi.org/10.1007/s10495-016-1234-5

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