Sulforaphane regulates macrophage glycolysis and inhibits the progression of diabetic nephropathy by modulating the ALOX5/NF-κB signaling pathway
10.19405/j.cnki.issn1000-1492.2024.03.004
- Author:
Rina Wu
1
;
Haidong Ding
1
;
Hong Chang
1
;
Nana Sun
1
;
Lei Zhang
1
Author Information
1. Dept of Endocrinology , Afiliated Hospital ofInner Mongolia University for Nationalities , Tongliao City , Inner Mongolia 028000
- Publication Type:Journal Article
- Keywords:
sulforaphane;
diabetic nephropathy;
macrophage;
glycolysis;
arachidonic acid 5-lipoxygenase;
NF- κB signaling pathway
- From:
Acta Universitatis Medicinalis Anhui
2024;59(3):390-397
- CountryChina
- Language:Chinese
-
Abstract:
Objective : To investigate the effects of sulforaphane (SFN) in regulating the macrophage glycolysis via the arachidonate 5-lipoxygenase (ALOX5) /nuclear factor kappa B (NF-κB) signaling pathway on the progression of diabetic nephropathy (DN) .
Methods :Bioinformatics analysis was used to identify the target genes of SFN in the treatment of DN . Human proximal tubular epithelial cell line (HK-2 cells) was induced with 30 mmol/L high glucose (HG) to create an in vitro model of DN . HK-2 cells were divided into the following groups : normal glucose (NG) group , HG group , HG + SFN (3 mmol/L) group , HG + ALOX5 group , HG + SFN (3 mmol/L) + ALOX5 group , HG-treated macrophages + HK-2 group , HG + SFN (3 mmol/L) treated macrophages s + HK-2 group , HG + ALOX5 transfection treated macrophages + HK-2 group , HG + SFN (3 mmol/L) + ALOX5 transfection treated macrophages + HK-2 group . CCK-8 assay was used to detect cell viability , Terminal deoxynucleotidyl transferase- mediated dUTP nick-end labeling (TUNEL) method was used to detect cell apoptosis; glucose and lactate levels in the cells were measured using assay kits; Western blot was performed to detect the expression of ALOX5 , NF-κB , and glycolysis-related proteins hexokinase-2 ( HK2 ) , pyruvate kinase M2 ( PKM2 ) , glucose transporter 1 (GLUT1) in each group . Diabetic nephropathy (DN) mouse models were established using streptozotocin (STZ) and treated with SFN (0. 5 mg/kg) . Various biochemical parameters were measured in the mice , and kidney tissue pathology was examined using H&E staining. Western blot was used to detect the expression of glycolysis-related proteins (HK2 , PKM2 , GLUT1) in kidney macrophages .
Results : Bioinformatics analysis revealed ALOX5 as the target gene of SFN in treating DN . Compared to the HG group , SFN treatment enhanced HK-2 cell viability and in- hibited apoptosis (P < 0. 05) ; concurrently , SFN treatment suppressed HG-induced macrophage glycolysis-related protein and attenuated macrophage-mediated HK-2 cellular injury ( P < 0. 05) . Western blot results showed that SFN inhibited the expression of ALOX5 and NF-κB ( P < 0. 05) . The mouse experiment results showed that SFN treatment improved kidney function and pathological changes in the kidney of DN mice , and inhibited the related protein expression of acrophage glycolysis in kidney tissue (P < 0. 05) .
Conclusion :SFN improves the progression of DN by inhibiting the expression of macrophage glycolysis-related protein through the ALOX5/NF-κB signaling pathway .
- Full text:2024062420301962176萝卜硫素通过调节ALOX5...胞糖酵解抑制糖尿病肾病进展_乌日娜.pdf
