The effect of conditional knockout of N-acetyltransferase 10 in renal tubular epithelial cells on the postnatal kidney of mice
Objective Explore the role of N-acetyltransferase 10(NAT10)in the kidney.Methods Establish a mouse model with conditional knockout of NAT10 in renal tubular epithelial cells at birth.The animals were divided into the control group and the group with conditional knockout of NAT10 in renal tubular epithelial cells(CKO group).Monitor the changes in body weight and serum creatinine in the two groups.Use pathological staining and electron microscopy to observe the morphological changes of renal tissues.Employ techniques such as Western blotting,immunofluorescence(IF),and immunohistochemical staining(IHC)to detect the impact of NAT10 on the markers of renal tubular and podocyte injury.Use Western blotting to detect the changes in inflammatory indicators.The unpaired t-test was used for compari-son between the two groups.Results The body weight of mice in CKO group was decreased[(23.35±2.82)g vs.(15.4±2.95)g,t=5.074,P＜0.05],and the creatinine level increased[(27.57±8.92)µmol/L vs.(53.35±7.28)μmol/L,t=5.570,P＜0.05].Pathological results showed that the renal tubules were dilated and there was infiltration of inflammatory cells in the renal interstitium.Western blotting results indicated that the expressions of Kidney injury molecule-1(KIM-1)(gray value ratio:1.000±0.045 vs.4.152±1.176,t=5.989,P＜0.05)and neutrophil gelatinase-associated lipocalin(NGAL)(gray value ratio:1.000±0.463 vs.3.045±0.233,t=8.815,P＜0.05)were elevated.IHC showed that the expression of KIM-1 in CKO group was significantly increased.Electron microscopy results revealed that the microvilli of renal tubules were broken and the mitochondrial structure of renal tubules was damaged.In addition,the expression of the podocyte marker Nephrin(gray value ratio:1.000±0.079 vs.0.525±0.166,t=5.780,P＜0.05)was decreased.IF showed that the expression of Wilms'tumor gene(WT-1)in CKO group was reduced.Electron microscopy results demonstrated that the foot processes of podocytes were fused and disappeared.The expressions of the inflammatory marker protein NOD-like recep-tor thermal protein domain associated protein 3(NLRP3)(gray value ratio:1.000±0.293 vs.1.986±0.483,t=3.902,P＜0.05)and interleukin-1β(IL-1β)(gray value ratio:1.000±0.250 vs.1.890±0.221,t=5.963,P＜0.05)were increased.Conclusion Conditional knockout of NAT10 in renal tubu-lar epithelial cells can induce injury to renal tubules and podocytes.
NETL
NSTL
万方数据
