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. 2019 May 14;55(1):103–115. doi: 10.3892/ijo.2019.4800

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Copyright: © Ji et al.

This is an open access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.

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Overexpression of Sirt6 in macrophages regulates the expression of apoptosis-related genes and Sirt6 in podocytes under high-glucose conditions. The diabetic internal environment was mimicked by a Transwell co-culture system under high-glucose conditions to co-culture the macrophages and podocytes. The cells in this experiment were divided into 8 groups as follows: i) The NM + HG + Con group (control; 50 mmol/l of high-glucose medium in the upper chamber, and podocytes with high-glucose medium in the lower chamber); ii) the HG + Con group (macrophages with high-glucose medium in the upper chamber, and podocyte with high-glucose medium in the lower chamber); iii) the HG + NC + Con group (macrophages transfected with NC plasmid with high-glucose medium in the upper chamber, and podocytes with high-glucose medium in the lower chamber); iv) the HG + Sirt6 + Con group (macrophages transfected with Sirt6 overexpression plasmid with high-glucose medium in the upper chamber, and podocytes with high-glucose medium in the lower chamber); v) the M-Sirt6 group (high-glucose medium in the upper chamber, and podocytes transfected with Sirt6 overexpression plasmid with high-glucose medium in the lower chamber); vi) the HG + M-Sirt6 group (macrophages with high-glucose medium in the upper chamber, and podocytes transfected with Sirt6 overexpression plasmid with high-glucose medium in the lower chamber); vii) the HG + NC + M-Sirt6 group (macrophages transfected with NC plasmid with high-glucose medium in the upper chamber, and podocytes transfected with Sirt6 overexpression plasmid with high-glucose medium in the lower chamber); and viii) the HG + Sirt6 + M-Sirt6 (macrophages transfected with Sirt6 overexpression plasmid with high-glucose medium in the upper chamber, and podocytes transfected with Sirt6 overexpression plasmid with high-glucose medium in the lower chamber). RT-qPCR was performed to examine the effects of the overexpression of Sirt6 on the mRNA expression levels of (A) Bcl-2, (B) Bax and (C) Sirt6 in the Transwell co-culture system. (D) Western blot analysis was performed to examine the effects of the overexpression of Sirt6 on the protein expression levels of Bcl-2, Bax and Sirt6 in the Transwell co-culture system. (E) Relative protein levels are presented as bar diagrams. GAPDH was used as an internal control. Data are presented as the means ± SD from 3 independent experiments (^*P<0.05 and ^**P<0.01, compared with the NM + HG + Con group; ^{^^}P<0.01, compared with the HG+NC+Con group; ^##P<0.01, compared with the M-Sirt6 group; ^&&P<0.01, compared with the HG + M-Sirt6 group).