图1 AGEs诱导的肾小管上皮细胞HK-2的差异分析和基因集富集分析(GSEA)

Fig.1 Differential analysis and gene set enrichment analysis (GSEA) for HK-2 renal tubular epithelial cells induced by advanced glycation end products. A: Heat map of analysis of variance between shCtrl-Normal and shCtrl-AGEs groups. B: Volcano plots of shCtrl-Normal and shCtrl-AGEs groups in analysis of variance. Blue dots represent low expression and red dots represent high expression. C: The top 3 terms shown in GSEA.

图2 基因本体论(GO)和京都基因和基因组百科全书(KEGG)

Fig.2 Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses. A: Biological process (BP) functional enrichment in GO analysis. B: Cell component (CC) functional enrichment in GO analysis. C: Molecular function (MF) function enrichment in GO analysis. D: Functional enrichment in WEGO analysis. E: Pathway enrichment in KEGG analysis.

图3 免疫浸润分析

Fig.3 Immune infiltration analysis. A: Bar graph of the proportions of 22 immune cells. B: Box-and-line plot showing comparison of the proportions of 22 immune cells in DKD group versus control group. C: Correlation of immune cells with 4 key genes.

图4 各组小鼠血清GSP、Scr、BUN和肾脏组织糖原沉积比较

Fig.4 Serum GSP, Scr, and BUN levels and renal glycogen deposition in mice in each group. A: Effect of QISD on glycated serum proteins in DKD mice (n=3). B: Effect of QISD on serum creatinine in DKD mice (n=3). C: Effect of QISD on serum urea nitrogen in DKD mice (n=3). D: PAS staining of the renal tissues of the mice (Original magnification: ×200) (n=6). ###P<0.001 vs control group; ***P<0.001 vs model group.

图5 各组小鼠肾脏组织病理变化比较

Fig.5 Histopathological changes in the kidneys of the mice in each group (×200) (n=6). A: HE staining. B: Masson staining.

图6 各组小鼠肾脏组织炎症因子表达量的比较

Fig.6 Expression of inflammatory factors in the kidney tissues of the mice in each group. A: Western blotting of TNF-α and MCP-1 proteins in renal tissues of the mice in each group. B, C: Quantitative analysis of the expression levels of TNF-α and MCP-1 proteins (n=3). D, E: RT-qPCR for detecting TNF-α and MCP-1 mRNA expressions in the renal tissues of the mice in each group (n=3). #P<0.05, ###P<0.001 vs control group; *P<0.05, **P<0.01, ***P<0.001 vs model group.

图7 各组小鼠肾脏组织KDM3C和SP1表达量的比较

Fig.7 Expression of KDM3C and SP1 in the kidney tissues of the mice in each group. A: Immunohistochemical staining for detecting KDM3C expression in the kidney tissues of the mice in each group (×200) (n=3). B: Western blotting for detecting protein expressions of KDM3C and SP1 in the kidney tissues of the mice in each group. C, D: Quantitative analysis of the expression levels of KDM3C and SP1 proteins (n=3). E, F: RT-qPCR for detecting expressions of KDM3C and SP1 mRNA in the kidney tissues of the mice in each group (n=3). #P<0.05, ###P<0.001 vs control group; *P<0.05, **P<0.01, ***P<0.001 vs model group.

图8 QISD对LPS诱导的HK-2细胞炎症损伤的影响

Fig.8 Effect of QISD on LPS-induced inflammatory damage in HK-2 cells. A: Viability of HK-2 cells after treatment with different LPS concentrations (50, 100, 200, 300, and 400 μg/mL) for 24, 48, and 72 h (n=6) determined by CCK8 assay. B-E: RT-qPCR for detecting mRNA expressions of KDM3C, SP1, TNF-α and MCP-1 in HK-2 cells (n=3). #P<0.05, ##P<0.01, ###P<0.001 vs control group; *P<0.05, **P<0.01, ***P<0.001 vs model group.

图9 在JIB-04作用下SP1、TNF-α、MCP-1和ICAM-1的表达水平变化

Fig.9 Changes in expression levels of SP1, TNF-α, MCP-1 and ICAM-1 in LPS-induced HK-2 cells after treatment with JIB-04. A: Western blotting protein bands of SP1 in HK-2 cells. B-E: RT-qPCR for detecting mRNA expressions of SP1, TNF-α, MCP-1 and ICAM-1 in HK-2 cells (n=3). #P<0.05, ##P<0.01, ###P<0.001 vs control group; *P<0.05, ***P<0.001 vs model group.