Profilin1 activates autophagy to participate in apoptosis of renal tubular epithelial cells induced by oxalate
Objective To investigate the effect of Profilin 1(PFN1)activation of autophagy in-volved in oxalic acid-mediated apoptosis of renal tubular epithelial cells.Methods Thirty patients di-agnosed with kidney stones in our hospital from April 2022 to July 2022 were selected as the stone group,and 30 patients without kidney stones were selected as the control group during the same peri-od.Urine specimens were collected from the study subjects of the two groups,and the concentration of PFN1 in the urine of the two groups was detected by ELISA.HK-2 cells were used as in vitro study subjects,which were divided into cell control group(basal medium),oxalic acid group(2 mmol/L oxalic acid),negative control group(2 mmol/L oxalic acid+siRNA negative control),si-PFN1 group(2 mmol/L oxalic acid+si-PFN1),and si-PFN1+rapamycin group(2 mmol/L oxalic acid+si-PFN1+rapamycin).Intracellular PFN1 expression was detected by qPCR and protein immunoblotting assay,and then PFN1 knockdown was performed in HK-2 cells using genetic engineering technology,and the Caspase3 enzyme activity content,mitochondrial membrane potential level,and the expression of the autophagy marker molecule Beclin1 were further detected in each group.Rescue experiments on the effect of PFN1 by drug intervention were performed,and the content of Caspase3 enzyme activity and the level of mitochondrial membrane potential were again detected in each group.Results The concentration of PFN1 in the urine of patients in the stone group was significantly higher than that in the control group(501.83±81.40 vs.236.67±77.09,P＜0.05).After siRNA knockdown of PFN1,PFN1 mRNA(1.07±0.21 vs.0.45±0.91)and protein expression(0.27±0.05 vs.0.16±0.03)were increased in the oxalic acid group compared with the cellular control group(all P＜0.05).Caspase3 enzyme activity was increased and mitochondrial membrane potential levels were decreased in the oxalic acid group compared to the cellular control group.Caspase3 enzyme activity was decreased in the si-PFN1 group and mitochondrial membrane potential level was increased compared to the nega-tive control group.Beclin1 protein and mRNA expression was increased in the oxalic acid group com-pared with the cellular control group(1.06±0.16 vs.0.28±0.07,P＜0.05).Beclin1 protein and mRNA expression was decreased in the si-PFN1 group compared with the negative control group(0.69±0.14 vs.1.13±0.11,P＜0.05).After the use of autophagy activator,Caspase3 enzyme ac-tivity was increased and mitochondrial membrane potential was decreased in the si-PFN1+rapamycin group compared with the si-PFN1 group.Conclusions PFN1 is involved in oxalic acid-mediated apop-tosis of renal tubular epithelial cells,which may result from the activation of autophagy,and PFN1 is expected to serve as a target for the prevention and treatment of renal stones.
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