Effects of sulforaphane on mitophagy-related proteins in nanobacteria-induced renal tubular epithelial cell apoptosis

doi:10.3969/j.issn.2095-4344.1484

Abstract

Abstract:

BACKGROUND: Sulforaohane has been shown to effectively improve the oxidative stress injury of renal tubular epithelial cells and inhibit the cell apoptosis to protect against renal tubular epithelial cell damage. However, the effect of sulforaohane on the mitophagy in renal tubular epithelial cells needs to be further studied.

OBJECTIVE: To explore the effects of sulforaphane on apoptosis- and mitophagy-related proteins in nanobacteria-induced renal tubular epithelial cell (HK-2) apoptosis.

METHODS: The study was in accordance with the ethics requirements of the Central Hospital of Enshi Tujia and Miao Autonomous Prefecture. Nanobacteria were from the urine of patients with kidney stone receiving no administration or undergoing surgery at Department of Urology, the Central Hospital of Enshi Tujia and Miao Autonomous Prefecture, and the patients signed the informed consents. HK-2 cells were divided into three groups, control group, nanobacteria-infected group, and sulforaphane intervention group (treated with nanobacteria and 30 μmol/L sulforaphane). The relative survival rates of HK-2 cells were determined using cell counting kit-8. Flow cytometry was used to detect the apoptotic rate of HK-2 cells. The expression levels of apoptosis-related proteins (Bax and Bcl-2) and proteins including PINK1, Parkin, LC3-II, LC3-I associated with mitophagy mediated by phosphatase and tensin homolog deleted on chromosome ten induced kinase 1 (PINK1)/Parkin were detected by western blot assay.

RESULTS AND CONCLUSION: (1) The inhibition of nanobacteria-induced cell proliferation was effectively lightened by sulforaphane (P < 0.05). (2) The expression levels of pro-apoptotic protein Bax and mitophagy-associated proteins PINK1 and Parkin and the ratio of LC3-II/LC3-I in HK-2 cells were up-regulated by nanobacteria (P < 0.05), and then were significantly down-regulated by sulforaphane (P < 0.05). The expression levels of anti-apoptotic protein Bcl-2 in HK-2 cells were down-regulated by nanobacteria (P < 0.05), and then was significantly up-regulated by sulforaphane (P < 0.05). (3) In summary, sulforaphane can inhibit the apoptosis of nanobacteria-induced renal tubular epithelial cells HK-2, and alleviate PINK1/Parkin-mediated mitophagy.

Key words: sulforaphane, renal tubular epithelial cells, nanobacteria, mitophagy, apoptosis, Bax, Bcl-2, LC3-II, LC3-I

CLC Number:

Pu Daojing, Zheng Fu, Xu Xianshun, Ding Wenjuan, Chen Yan, Zou Zhaoyin. Effects of sulforaphane on mitophagy-related proteins in nanobacteria-induced renal tubular epithelial cell apoptosis[J]. Chinese Journal of Tissue Engineering Research, 2019, 23(34): 5503-5507.

Figures/Tables 4

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