Abstract
Objective To investigate the regulatory effects of paeonol on autophagy and the NOD-like receptor protein 3(NLRP3)inflammasome in human renal proximal tubular epithelial cells(HK-2)induced by human serum albumin(HSA).Methods HK-2 cells were cultured in vitro and randomly assigned to control group,model group,as well as low-,medium-and high-dose experimental groups.Cells in the control group were cultured under normal conditions;those in the model group were treated with 20 mg·mL-1 HSA for 24 h;and cells in the low-,medium-and high-dose experimental groups were administered 25,50 and 100 μmol·L-1 paeonol,respectively,for 24 h on the basis of the model group intervention.Enzyme-linked immunosorbent assay(ELISA)was performed to determine the concentrations of kidney injury molecule 1(KIM-1),monocyte chemoattractant protein 1(MCP-1),interleukin 1 β(IL-1β)and caspase-1 in the cell supernatant.Quantitative real-time polymerase chain reaction(qRT-PCR)was used to detect the mRNA expression levels of KIM-1 and MCP-1.Western blotting was applied to measure the protein expression levels of NLRP3,apoptosis-associated speck-like protein containing a CARD(ASC),interleukin-1 β(IL-1 β),microtubule-associated protein 1 light chain 3(LC3),sequestosome 1(p62),Beclin-1,PTEN-induced kinase 1(PINK1)and Parkin.Results The concentrations of KIM-1 in the cell supernatant of the control group,model group,and low-,medium-and high-dose experimental groups were(8.65±0.53),(17.33±2.04),(10.76±1.41),(8.48±0.47)and(8.47±0.24)pg·mL-1,respectively;the contents of MCP-1 were(531.38±15.64),(2 198.55±148.21),(1 966.29±53.52),(1 605.49±234.68)and(937.33±36.96)pg·mL-1,respectively;the secretion levels of IL-1 β were(71.66±2.29),(109.92±4.41),(91.68±4.75),(80.66±4.00)and(62.38±3.24)pg·mL-1,respectively;the contents of caspase-1 were(66.57±10.52),(210.50±17.50),(201.88±7.22),(195.42±13.95)and(129.81±13.20)pg·mL-1,respectively;the relative expression levels of KIM-1 mRNA in each group were 1.05±0.05,2.38±0.45,1.58±0.20,1.44±0.08 and 1.38±0.22,respectively;while those of MCP-1 were 1.04±0.04,1.53±0.08,1.32±0.16,1.16±0.16 and 0.62±0.13,respectively;the relative expression levels of NLRP3 protein were 0.89±0.08,1.13±0.07,1.00±0.07,0.84±0.12 and 0.55±0.16,respectively;those of ASC were 0.23±0.03,0.87±0.19,0.81±0.16,0.78±0.08 and 0.46±0.05,respectively;those of IL-1 β were 0.49±0.12,1.46±0.29,1.10±0.26,0.71±0.06 and 0.53±0.13,respectively;the LC3 Ⅱ/Ⅰ ratios were 1.08±0.04,0.90±0.06,1.00±0.02 and 1.05±0.04,respectively;the expression levels of p62 protein were 0.38±0.06,1.23±0.11,1.05±0.20 and 0.62±0.19,respectively;those of Beclin-1 were 0.82±0.07,0.59±0.06,0.90±0.07 and 0.93±0.09,respectively;those of PINK1 were 1.10±0.03,0.68±0.07,0.78±0.11 and 1.03±0.08,respectively;those of Parkin were 0.64±0.06,0.46±0.05,0.58±0.08 and 0.79±0.12,respectively.Compared with the control group,all the above indicators in the model group exhibited statistically significant differences(P＜0.05,P＜0.01,P＜0.001).Moreover,the indicators in the high-dose experimental group showed statistically significant differences compared with the model group(P＜0.05,P＜0.01,P＜0.001).Conclusion Paeonol may alleviate albumin-induced renal tubular injury by promoting mitophagy and inhibiting the activation of the NLRP3 inflammasome.关键词
丹皮酚/白蛋白/自噬/炎症小体/肾小管上皮细胞Key words
paeonol/albumin/autophagy/inflammasome/renal tubular epithelial cell分类
医药卫生
