Nephroprotective mechanism of Kunkui Baoshen decoction in diabetic kidney disease: Targeting the HERC2/NCOA4-mediated autophagy-dependent ferroptosis pathway

Figure 4 Kunkui Baoshen decoction alleviates inflammation, fibrosis, and cellular apoptosis in high glucose-induced human kidney-2 cells. A: Human kidney-2 (HK-2) cell viability under different glucose concentrations compared to the normal group (NG, 5.5 mmol/L glucose); B: HK-2 cell viability under high glucose (HG, 50 mmol/L glucose) with varying concentrations of Kunkui Baoshen decoction; C: Morphological changes of HK-2 cells observed at 4 × and 40 × magnification (scale bar = 500 μm); D: Wound healing assay to evaluate HK-2 cell migration under different treatments (scale bar = 500 μm); E: Quantification of migration rate; F-I: Western blot and quantitative reverse transcription-polymerase chain reaction (qRT-PCR) analyses of cysteine-aspartic acid protease-3 and Bcl-2 associated X protein in HK-2 cells; J-M: Western blot and qRT-PCR analyses of interleukin-6 and tumor necrosis factor-α in HK-2 cells; N-R: Western blot and qRT-PCR analyses of epithelial-cadherin, α-smooth muscle actin, and transforming growth factor-β1 in HK-2 cells. NG: Normal group; Man: Mannitol (osmotic control); HG: High glucose; KKBS: Kunkui Baoshen decoction; Caspase-3: Cysteine-aspartic acid protease-3; Bax: Bcl-2 associated X; GAPDH: Glyceraldehyde 3-phosphate dehydrogenase; IL-6: Interleukin-6; TNF-α: Tumor necrosis factor-α; E-cadherin: Epithelial-cadherin; TGF-β1: Transforming growth factor-β1; α-SMA: α-smooth muscle actin. ^aP < 0.05 vs normal group, ^bP < 0.01 vs normal group, ^cP < 0.001 vs normal group, ^dP < 0.0001 vs normal group; ^eP < 0.05 vs high glucose group, ^fP < 0.01 vs high glucose group, ^gP < 0.001 vs high glucose group, ^hP < 0.0001 vs high glucose group.