Effects of Overexpression of SNHG5 on High Glucose-Induced Apoptosis and Oxidative Stress in HK-2 Cells
HU Wei1, DAI Qiong1, DING Guimei1, LIU Xiang1, HUA Yan2*, SHENG Shangchun1
This study investigated the effect and molecular mechanism of SNHG5 (small nucleolar RNA host gene 5) on HG (high glucose)-induced HK-2 apoptosis and oxidative stress. HK-2 cells were treated with 30 mmol/L glucose for 48 h. pcDNA, pcDNA-SNHG5, anti-miR-NC (negative control), microRNA-196a (miRNA/miR-196a) inhibitor, pcDNA-SNHG5 were co-transfected with miR-NC or miR-196a mimic into HK-2 cells, respectively. The expression levels of SNHG5 and miR-196a were detected by RT-qPCR (quantitative real-time PCR); cell proliferation was detected by CCK-8 (cell counting kit-8); flow cytometry was performed to determine cell apoptosis; protein expression detected by Western blot; MDA (malondialdehyde), SOD(superoxide dismutase), glutathione peroxidase GSH-Px (glutathione peroxidase) kits were implemented to monitor the content of MDA and the activities of SOD and GSH-Px; the targeting relationship between SNHG5 and miR-196a was detected by dual-luciferase reporter assay. The study concluded that the expression level of SNHG5, cell viability, and expression level of Bcl2 (B cell lymphoma/lewkmia-2) in HK-2 cells induced by high glucose decreased, miR-196a expression level, apoptosis rate, expression of activated Cleaved-caspase3 (Cleaved cysteinyl aspartate specific proteinase 3) and Bax (Bcl-2 associated X protein) level and MDA content increased, while SOD and GSH-Px activities decreased (P<0.05). After overexpression of SNHG5 or inhibition of miR-196a expression, high glucose-induced HK-2 cell activity significantly increased and cell death rate decreased, Bcl-2 expression level increased significantly, Bax, Cleaved-caspase3 and MDA contents significantly decreased, SOD, GSH-Px activity increased (P<0.05). SNHG5 can target and regulate miR-196a expression, and miR-196a overexpression can reverse the effects of SNHG5 on high glucose-induced HK-2 cell viability, apoptosis and oxidative stress. In conclusion, overexpression of SNHG5 may inhibit high glucose-induced HK-2 apoptosis and oxidative stress by downregulating miR-196a.
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