【摘要】 目的 探究转运RNA衍生片段003634(tRNA-derived fragment 003634,tRF-003634)对阿霉素(adriamycin)肾病小鼠足细胞凋亡的作用及机制。方法 (1)用BALB/c小鼠作为对象来建立阿霉素(10 mg/kg)肾病模型。取20只BALB/c小鼠随机分为：对照组、阿霉素组(ADR组)、ADR+tRF-003634 agomir组、ADR+tRF-003634 NC组(n=5)。检测各组小鼠的血清生化指标水平，H-E染色、PAS染色评估小鼠肾脏病理变化；Real-Time PCR法检测各组肾脏中tRF-003634的表达水平；Western blot法用来检验肾脏组织中caspase3、cleaved caspase3和p53的蛋白表达水平。(2)体外用阿霉素(1 mg/L)诱导足细胞凋亡。依据不同干预因素将足细胞分为：对照组、ADR组、ADR+tRF-003634 mimics组和ADR+tRF-003634 NC组。Real-Time PCR法测定各组tRF-003634的表达水平；Western blot法用来测定不同组别间足细胞中caspase3、cleaved caspase3和p53的相对表达。(3)将足细胞分成：对照组、ADR组、ADR+SB203580组和SB203580组。利用Western blot法来检测每组p-p38MAPK和p-Hsp27的蛋白相对表达，Real-time PCR法检测tRF-003634的表达水平。结果 (1)与对照组相比，ADR组的病理变化显著加重，tRF-003634的表达水平下降，cleaved caspase3和p53的表达水平显著上升(P<0.05)。与ADR组相比，ADR+tRF-003634 agomir组小鼠24 h尿蛋白和血清尿素氮水平显著下降(P<0.05),病理改变减轻；tRF-003634表达水平明显上升，cleaved caspase3和p53表达水平显著下降(P<0.05)。(2)与对照组相比，ADR组足细胞中tRF-003634表达水平显著下降，cleaved caspase3和p53表达水平显著上升(P<0.05)。与ADR组相比，ADR+tRF-003634 mimics组的tRF-003634表达水平显著上升(P<0.05),cleaved caspase3和p53表达水平显著下降(P<0.05)。(3)与对照组相比，ADR组p-p38MAPK和p-Hsp27表达上调，tRF-003634表达下降(P<0.05);与ADR组相比，ADR+SB203580组p-p38MAPK和p-Hsp2蛋白表达显著下降，tRF-003634表达上升(P<0.05)。结论 tRF-003634能够抑制足细胞凋亡，减轻阿霉素诱导的肾脏病理损伤。tRF-003634可能作为MAPK通路的下游效应分子改善足细胞凋亡，为慢性肾脏病的诊治提供新的思路。更多还原

【Abstract】 Objective To explore the effect of tRF-003634(tRNA-derived fragment 003634) on podocyte apoptosis in mice with adriamycin nephropathy and related mechanism.Methods(1)A model of adriamycin(10 mg/kg) nephropathy was established using BALB/c mice. 20 BALB/c mice were divided randomly into four groups(n=5): a control group, an ADR group, an ADR+tRF-003634 agomir group and an ADR+tRF-003634 NC group. Their serum biochemical indexes were detected, and the pathological changes in the kidneys were evaluated by HE and PAS staining. The expression of tRF-003634 in each group was detected by real-time PCR. The expression of caspase3, cleaved caspase3 and p53 was detected by Western blot.(2)Adriamycin(1 mg/L) was used to induce podocyte apoptosis in vitro. According to different intervention factors, podocytes were divided into four groups: a control group, an ADR group, an ADR+tRF-003634 mimics group and an ADR+tRF-003634 NC group. The expression of tRF-003634 was detected by real-time PCR. The relative expression of caspase3, cleaved caspase3 and p53 in podocytes was examined by Western blot.(3)Podocytes were divided into four groups: a control group, an ADR group, an ADR+SB203580 group and a SB203580 group. The expression of p-p38MAPK and p-Hsp27 was measured by Western blot, and the expression of tRF-003634 was detected by real-time PCR. Results(1)Compared with the control group, the ADR group showed significantly increased pathological changes, as well as decreases in the expression of tRF-003634 and increases in the expression of cleaved caspase3 and p53(P<0.05). Compared with the ADR group, mice in the ADR+tRF-003634 agomir group presented significantly reduced 24 h urine protein and serum urea nitrogen(P<0.05), with alleviated pathological changes, as well as increase in the expression of tRF-003634 decreases in the expression of cleaved caspase3 and p53(P<0.05).(2)Compared with the control group, the ADR group presented significant decreases in tRF-003634 expression, and increases in the levels of cleaved caspase3 and p53(P<0.05). Compared with the ADR group, the tRF-003634 mimics group showed remarkably increased expression of tRF-003634(P<0.05), and decreased expression of cleaved caspase3 and p53(P<0.05).(3)Compared with the control group, the expression of p-p38MAPK and p-Hsp27 in the ADR group was up-regulated, and the expression of tRF-003634 was down-regulated(P<0.05). Compared with the ADR group, the expression of p-p38MAPK and p-Hsp27 in the ADR+SB203580 group significantly decreased, and the expression of tRF-003634 increased(P<0.05). Conclusions tRF-003634 can inhibit the apoptosis of podocytes and alleviate renal pathological damage induced by adriamycin. tRF-003634 may act as a downstream effector molecule of MAPK pathway to improve podocyte apoptosis, providing new thoughts for the diagnosis and treatment of chronic renal disease.更多还原