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. 2023 Mar 15;10:1151359. doi: 10.3389/fmed.2023.1151359

Figure 4.

Figure 4

The effect of acrolein scavengers on Acr-PC formation and PKM2 activity in kidney tissues of HFD-STZ-induced DN mice. Male 6-week-old C57BL/6 J mice were fed a normal diet or a high-fat diet (HFD) for 16 weeks. In the DN group, the procedure was performed as described in Figure 1. In the DN+ acrolein scavenger group, DN mice were treated with vehicle (HF, n = 5) or acrolein scavengers, including NAC (HF + NAC, 1 g/kg/d in drinking water, n = 5), hydralazine (HF + Hyl, 50 mg/kg/d in drinking water, n = 5), or carnosine (HF + Car, 50 mg/kg/d in drinking water, n = 5) for 16 weeks. In the acrolein scavenger group, normal diet-fed mice were treated with acrolein scavengers as described above (n = 4 for each scavenger, CTR + NAC, CTR+ Hyl, CTR + Car). Control mice (CTR, n = 5) were fed a normal diet and received vehicle administration. (A) Western blot analysis of Acr-PC levels in kidney tissues of the control group (CTR, n = 5), DN group (HF, n = 5), DN + acrolein scavenger group (n = 5 for each scavenger, HF + NAC, HF + Hyl, HF + Car) and acrolein scavenger group (n = 4 for each scavenger, CTR + NAC, CTR + Hyl, CTR + Car). (B) Quantification of (A). (C) The left panel shows a representative blot image of cross-linked mouse kidney samples showing PKM2 monomers, dimers, and tetramers. The right panel shows the ratio between tetramer and dimer + monomer of PKM2 in kidney samples of the control group (n = 5), DN group (n = 5), DN+ acrolein scavenger group and acrolein scavenger group (n = 5). (D) Analysis of PK activity in kidney tissues of the control group (n = 5), DN group (n = 5), DN+ acrolein scavenger group (n = 5) and acrolein scavenger group (n = 4). The values are presented as the mean ± SD. Kruskal-Wallis tests were used to determine statistical significance, and two-tailed p values are shown. *p < 0.05, **p < 0.01 compared with the control group. #p < 0.05 compared with the DN group.