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果糖通过尿酸和活性氧簇诱导人肾小管上皮细胞单核细胞趋化蛋白-1的表达 被引量:1
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作者 王巧玲 陈晓欢 +3 位作者 倪兆慧 顾乐怡 徐辰祺 戴慧莉 《上海交通大学学报(医学版)》 CAS CSCD 北大核心 2018年第4期386-393,共8页
目的·探讨果糖诱导肾小管上皮细胞表达单核细胞趋化蛋白-1(monocyte chemoattratant protein-1,MCP-1)的机制。方法·将HK-2细胞分为对照组,果糖孵育(1、5和10 mmol/L)组,果糖、己酮糖激酶抑制剂(KHK-IN)共孵育(果糖5 mmol/L,K... 目的·探讨果糖诱导肾小管上皮细胞表达单核细胞趋化蛋白-1(monocyte chemoattratant protein-1,MCP-1)的机制。方法·将HK-2细胞分为对照组,果糖孵育(1、5和10 mmol/L)组,果糖、己酮糖激酶抑制剂(KHK-IN)共孵育(果糖5 mmol/L,KHKIN分别为12、100和1 000 nmol/L)组,尿酸孵育(5、15和50 mg/d L)组,果糖、别嘌醇共孵育(果糖5 mmol/L,别嘌醇分别为0.01、0.1和0.5 mmol/L)组,尿酸、别嘌醇共孵育(尿酸50 mg/d L,别嘌醇分别为0.01、0.1和0.5 mmol/L)组,H2O2孵育(0.1和0.3 mmol/L)组,果糖、N-乙酰半胱氨酸(NAC)共孵育(果糖5 mmol/L,NAC分别为5、10和50 mmol/L)组,尿酸、NAC共孵育(尿酸50 mg/d L,NAC分别为5、10和50 mmol/L)组。采用实时荧光定量PCR和蛋白免疫印迹实验检测MCP-1 m RNA和蛋白的表达;利用活性氧(ROS)荧光探针观察果糖和尿酸对HK-2细胞ROS产生的影响。结果·果糖剂量及时间依赖性诱导HK-2细胞MCP-1 m RNA转录和蛋白表达,此过程可被KHK-IN阻断。外源性尿酸诱导HK-2细胞产生MCP-1,别嘌醇抑制果糖引起的MCP-1表达,但不能阻断外源性尿酸的作用。果糖和尿酸均诱导HK-2细胞产生ROS,别嘌醇抑制了果糖而非外源性尿酸诱导的ROS产生。H2O2诱导HK-2细胞生成MCP-1,NAC则抑制这种作用。结论·果糖在己酮糖激酶催化下,通过引起细胞内尿酸升高和产生ROS,导致人肾小管上皮细胞产生MCP-1。 展开更多
关键词 果糖 尿酸 活性氧簇 HK-2细胞
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Advanced wasting in peritoneal dialysis patients 被引量:1
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作者 Zhi Xu Glen H Murata +4 位作者 Robert H Glew Yijuan Sun Darlene Vigil Karen S Servilla Antonios H Tzamaloukas 《World Journal of Nephrology》 2017年第3期143-149,共7页
AIM To identify patients with end-stage renal disease treated by peritoneal dialysis(PD)who had zero body fat(BF)as determined by analysis of body composition using anthropometric formulas estimating body water(V)and ... AIM To identify patients with end-stage renal disease treated by peritoneal dialysis(PD)who had zero body fat(BF)as determined by analysis of body composition using anthropometric formulas estimating body water(V)and to compare nutritional parameters between these patients and PD patients whose BF was above zero.METHODS Body weight(W)consists of fat-free mass(FFM)andBF.Anthropometric formulas for calculating V allow the calculation of FFM as V/0.73,where 0.73 is the water fraction of FFM at normal hydration.Wasting from loss of BF has adverse survival outcomes in PD.Advanced wasting was defined as zero BF when V/0.73 is equal to or exceeds W.This study,which analyzed 439 PD patients at their first clearance study,used the Watson formulas estimating V to identify patients with V_(Watson)/0.73≥W and compared their nutritional indices with those of PD patients with V_(Watson)/0.73<W.RESULTS The study identified at the first clearance study two male patients with V_(Watson)/0.73≥W among 439 patients on PD.Compared to 260 other male patients on PD,the two subjects with advanced wasting had exceptionally low body mass index and serum albumin concentration.The first of the two subjects also had very low values for serum creatinine concentration and total(in urine and spent peritoneal dialysate)creatinine excretion rate while the second subject had an elevated serum creatinine concentration and high creatinine excretion rate due,most probably,to non-compliance with the PD prescription.CONCLUSION Advanced wasting(zero BF)in PD patients,identified by the anthropometric formulas that estimate V,while rare,is associated with indices of poor somatic and visceral nutrition. 展开更多
关键词 Weight deficit Fat-free mass Nutrition Body water ANTHROPOMETRY Peritoneal dialysis Watson formulas WASTING
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Nephropathy in dietary hyperoxaluria:A potentially preventable acute or chronic kidney disease 被引量:4
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作者 Robert H Glew Yijuan Sun +5 位作者 Bruce L Horowitz Konstantin N Konstantinov Marc Barry Joanna R Fair Larry Massie Antonios H Tzamaloukas 《World Journal of Nephrology》 2014年第4期122-142,共21页
Hyperoxaluria can cause not only nephrolithiasis and nephrocalcinosis,but also renal parenchymal disease histologically characterized by deposition of calcium oxalate crystals throughout the renal parenchyma,profound ... Hyperoxaluria can cause not only nephrolithiasis and nephrocalcinosis,but also renal parenchymal disease histologically characterized by deposition of calcium oxalate crystals throughout the renal parenchyma,profound tubular damage and interstitial inflammation and fibrosis.Hyperoxaluric nephropathy presents clinically as acute or chronic renal failure that may progress to endstage renal disease(ESRD).This sequence of events,well recognized in the past in primary and enteric hyperoxalurias,has also been documented in a few cases of dietary hyperoxaluria.Estimates of oxalate intake in patients with chronic dietary hyperoxaluria who developed chronic kidney disease or ESRD were comparable to the reported average oxalate content of the diets of certain populations worldwide,thus raising the question whether dietary hyperoxaluria is a primary cause of ESRD in these regions.Studies addressing this question have the potential of improving population health and should be undertaken,alongside ongoing studies which are yielding fresh insights into the mechanisms of intestinal absorption and renal excretion of oxalate,and into the mechanisms of development of oxalate-induced renal parenchymal disease.Novel preventive and therapeutic strategies for treating all types of hyperoxaluria are expected to develop from these studies. 展开更多
关键词 Dietary hyperoxaluria Chronic oxalatenephropathy Acute oxalate nephropathy Acute tubular necrosis Interstitial nephritis NEPHROCALCINOSIS Calcium oxalate nephrolithiasis Oxalate transporters Inflammasomes
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Reproducibility of serial creatinine excretion measurements in peritoneal dialysis
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作者 ZhiXu Glen H Murata +6 位作者 Yijuan Sun Robert H Glew Clifford Qualls Darlene Vigil Karen SServilla Thomas A Golper Antonios H Tzamaloukas 《World Journal of Nephrology》 2017年第4期201-208,共8页
AIMTo test whether muscle mass evaluated by creatinine excretion (EXCr) is maintained in patients with end-stage kidney disease (ESKD) treated by peritoneal dialysis (PD), we evaluated repeated measurements of E... AIMTo test whether muscle mass evaluated by creatinine excretion (EXCr) is maintained in patients with end-stage kidney disease (ESKD) treated by peritoneal dialysis (PD), we evaluated repeated measurements of EXCr in a PD population.METHODSOne hundred and sixty-six PD patients (94 male, 72 female) receiving the same PD dose for the duration of the study (up to approximately 2.5 years) had repeated determinations of total (in urine plus spent dialysate) 24-h EXCr (EXCr T) to assess the adequacy of PD by creatinine clearance. All 166 patients had two EXCr T determinations, 84 of the 166 patients had three EXCr T determinations and 44 of the 166 patients had four EXCr T measurements. EXCr T values were compared using the paired t test in the patients who had two studies and by repeated measures ANOVA in those who were studied three or four times.RESULTSIn patients who were studied twice, with the first and second EXCr T measurements performed at 9.2 ± 15.2 mo and 17.4 ± 15.8 mo after onset of PD, respectively, EXCr T did not differ between the first and second study. In patients studied three times and whose fnal assessment occurred 24.7 ± 16.3 mo after initiating PD, EXCr T did not differ between the first and second study, but was significantly lower in the third study compared to the frst study. In patients who were studied four times and whose fourth measurement was taken 31.9 ± 16.8 mo after onset of PD, EXCr T did not differ between any of the studies. The average EXCr T value did not change signifcantly, with the exception of the third study in the patients studied thrice. However, repeated determinations of EXCr T in individuals showed substantial variability, with approximately 50% of the repeated determinations being higher or lower than the first determination by 15% or more.CONCLUSIONThe average value of EXCr T remains relatively constantfor up to 2.5 years of follow-up in PD patients who adhereto the same PD schedule. However, repeated individualEXCr T values vary considerably in a large proportion ofthe patients. Further studies are needed to evaluatethe clinical signifcance of varying EXCr T values and thestability of EXCr T beyond 2.5 years of PD follow-up. 展开更多
关键词 Creatinine excretion Peritoneal dialysis Lean body mass Muscle mass
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热休克蛋白72抑制ATP耗竭时细胞色素C释放所诱导的肾小管上皮细胞凋亡 被引量:3
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作者 毛海萍 余学清 +7 位作者 李志坚 杨琼琼 尹培达 李芳红 K.LRuchalski J.H.Schwartz S.C.Borkan 汪一汗 《中华肾脏病杂志》 CAS CSCD 北大核心 2003年第3期137-141,共5页
目的 研究热休克蛋白(HSP)72对ATP耗竭时细胞色素C释放所导致的肾小管上皮细胞凋亡的保护作用及其分子机制。方法 应用代谢抑制剂暂时性阻断细胞内ATP的生成,引起细胞凋亡。应用热处理细胞或编码HSP72 RNA的腺病毒感染细胞,诱导HSP72的... 目的 研究热休克蛋白(HSP)72对ATP耗竭时细胞色素C释放所导致的肾小管上皮细胞凋亡的保护作用及其分子机制。方法 应用代谢抑制剂暂时性阻断细胞内ATP的生成,引起细胞凋亡。应用热处理细胞或编码HSP72 RNA的腺病毒感染细胞,诱导HSP72的表达。以Western印迹检测释放于胞浆内的细胞色素C。荧光肽法测定半胱氨酸天冬氨酸蛋白酶(caspase)3活性。Hoechst33342染色观察细胞凋亡的发生情况。结果 肾小管上皮细胞内ATP耗竭时,释放至胞浆内的细胞色素C的含量增多,caspase 3活性增强;细胞内ATP再恢复时,细胞色素C的释放和caspase 3活性进一步增加,细胞体积缩小,核浓染、固缩,形成凋亡小体。预先热处理后,各组细胞色素C的释放明显减少,caspase 3活性显著抑制(P<0.05,n=3)。高表达HSP72时,各时间点caspase 3活性的抑制程度与热处理组相似,细胞体积缩小,核浓染、固缩,凋亡小体的形成明显减少。结论 HSP72可抑制ATP耗竭时细胞色素C所导致的肾小管上皮细胞凋亡,其机制是抑制凋亡通路中细胞色素C的释放和caspase 3的激活。 展开更多
关键词 热休克蛋白72 ATP 细胞色素C 肾小管上皮细胞 细胞凋亡 半胱氨酸天冬氨酸蛋白酶3
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囊泡型H^+-ATPase B1亚基的突变对大鼠内髓集合管细胞H^+-ATPase结构和泵氢功能的影响 被引量:3
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作者 杨琼琼 毛海萍 +2 位作者 李广木 John H Schwartz 余学清 《中华肾脏病杂志》 CAS CSCD 北大核心 2009年第1期24-30,共7页
目的研究致遗传性远端肾小管酸中毒(dRTA)的囊泡型H^+-ATPaseB1亚基(ATP6V181)的点突变对大鼠内髓集合管(IMCD)细胞H^+-ATPase结构和泵氢功能的影响。方法模拟致人类遗传性dRTA的B1亚基点突变构建野生型(WT)和7种突变型(M)... 目的研究致遗传性远端肾小管酸中毒(dRTA)的囊泡型H^+-ATPaseB1亚基(ATP6V181)的点突变对大鼠内髓集合管(IMCD)细胞H^+-ATPase结构和泵氢功能的影响。方法模拟致人类遗传性dRTA的B1亚基点突变构建野生型(WT)和7种突变型(M)质粒,转染大鼠IMCD细胞并筛选稳定表达绿色荧光蛋白(CFP)-B1 M和GFP—B1 WT的IMCD细胞系。应用免疫荧光、免疫蛋白印迹法、ATP。NADH耦合实验和快速酸负荷后不依赖钠的细胞内pH的变化,来观察GFP—B1 M和GFP—B1 WT在细胞内的分布,及其与H^+-ATPase其他(E、H和c)亚基结合能力对ATP酶活性和H^+-ATPase泵氢功能的影响。结果GFP.B1wT在转染细胞中呈囊泡样分布,与H^+-ATPase的分布一致;而GFP—B1 M则为弥散分布。免疫沉淀结果显示只有GFP—B1 WT融合蛋白能和其他的H^+-ATPase亚基(E、H和c)结合形成复合物,而GFP-B1 M融合蛋白无此作用。ATP酶活性只有在GFP—B1 WT转染细胞株的免疫沉淀产物中存在,在GFP—B1 M转染细胞株的免疫沉淀产物中不存在。在GFP—B1 M转染的IMCD细胞快速酸负荷后H^+-ATPase介导的钠不依赖pHi的恢复受到显著抑制[pHi的恢复率(pHU/min)在L81P、R124W、M174R、P275R、G316E、P346R、G364S GFP—B1M转染的IMCD细胞分别为0.007±0.002、0.004±0.002、0.002±0.002、0.003±0.002、0.006±0.004、0.009±O.004、0.015±0.006,P〈0.05,n=51。而GFP—B1 WT转染的IMCD细胞pHi的恢复率与未转染IMCD细胞相似[(0.040±0.006)pH U/min],且能被1μmol/L巴弗洛霉素(H^+-ATPase特异性抑制剂)所抑制。结论遗传性dRTA囊泡型H^+-ATPaseB1亚基点突变影响GFP—B1融合蛋白与其他亚基正常结合组装形成完整的H^+-ATPase,并抑制H^+-ATPase的泌酸功能。 展开更多
关键词 质子转运ATP酶类 酸中毒 肾小管性 点突变 囊泡型H^+- ATPASE B1亚基突变 内髓集合管细胞
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细胞色素C在ATP耗竭诱导的肾小管上皮细胞凋亡中的作用 被引量:3
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作者 毛海萍 余学清 +6 位作者 杨琼琼 尹培达 李芳红 K.LRuchalski J.H.Schwartz S.C.Borkan 汪一汗 《中华肾脏病杂志》 CAS CSCD 北大核心 2003年第2期90-94,共5页
目的 研究线粒体细胞色素C的释放在ATP耗竭导致的肾小管上皮细胞凋亡中的作用和机制。方法 应用代谢抑制剂暂时性阻断细胞内ATP的生成,诱导细胞凋亡。利用JC-1测定线粒体膜电位的变化。以间接免疫荧光和Western印迹检测细胞色素C在细胞... 目的 研究线粒体细胞色素C的释放在ATP耗竭导致的肾小管上皮细胞凋亡中的作用和机制。方法 应用代谢抑制剂暂时性阻断细胞内ATP的生成,诱导细胞凋亡。利用JC-1测定线粒体膜电位的变化。以间接免疫荧光和Western印迹检测细胞色素C在细胞内的分布。荧光肽法测定Caspase3的活性。琼脂糖凝胶电泳分析DNA碎解。结果 肾小管上皮细胞内ATP耗竭时,线粒体膜电位显著降低,细胞浆内细胞色素C的含量增多,Caspase 3活性增强。细胞内ATP再恢复时,细胞色素C的释放和Caspase 3活性继续增加,并出现DNA的碎解。结论 肾小管上皮细胞ATP耗竭时,线粒体细胞色素C的释放在介导Caspase依赖的细胞凋亡通路中发挥重要作用。 展开更多
关键词 肾小管上皮细胞 细胞凋亡 线粒体 细胞色素C CASPASE3
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肾小管上皮细胞ATP耗竭再恢复时热休克蛋白72与Paxillin的相互作用 被引量:1
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作者 毛海萍 李志坚 +3 位作者 余学清 J.H.Schwartz S.C.Borkan 汪一汗 《中华肾脏病杂志》 CAS CSCD 北大核心 2006年第7期411-415,共5页
目的研究肾小管上皮细胞ATP耗竭再恢复时热休克蛋白(HSP)72与桩蛋白(Paxillin)的相互作用和意义。方法应用代谢抑制剂暂时性阻断肾小管上皮细胞ATP的生成,引起细胞内的ATP耗竭;换用含10 mmol/L葡萄糖的DMEM培养液,使细胞内ATP再恢复;... 目的研究肾小管上皮细胞ATP耗竭再恢复时热休克蛋白(HSP)72与桩蛋白(Paxillin)的相互作用和意义。方法应用代谢抑制剂暂时性阻断肾小管上皮细胞ATP的生成,引起细胞内的ATP耗竭;换用含10 mmol/L葡萄糖的DMEM培养液,使细胞内ATP再恢复;以热处理细胞或编码HSP72 RNA的腺病毒感染细胞,诱导细胞高表达HSP72。Western印迹检测HSP72水平;间接免疫荧光和Western印迹检测Paxillin在细胞内的分布变化。免疫共沉淀观察HSP72与Paxillin的相互作用。结果肾小管上皮细胞ATP耗竭再恢复时,对照组细胞内的Paxillin由局部黏附结构区域向细胞浆内弥散分布;HSP72由细胞浆转移至细胞膜。细胞高表达HSP72后,HSP72在细胞浆和细胞膜中的蛋白含量均增加(P<0.05);Paxillin由细胞浆向细胞膜的转移明显减少(P<0.05);Paxillin在细胞内的正常分布改善;HSP72和Paxillin之间的相互作用显著增加(P<0.05)。结论肾小管上皮细胞ATP耗竭再恢复时,HSP72可保持Paxillin在细胞内的正常分布特点和区域,其机制可能是HSP72增加与Paxillin的相互作用,发挥分子伴侣的功能。 展开更多
关键词 热休克蛋白 桩蛋白 细胞骨架 再灌注损伤 腺苷三磷酸 肾小管 上皮细胞
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半胱氨酸天冬氨酸3依赖的bcl-2的降解在ATP耗竭诱导肾小管上皮细胞凋亡中的作用
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作者 李志坚 毛海萍 +3 位作者 余学清 汪一汗 J.H.Schwartz S.C.Borkan 《中华肾脏病杂志》 CAS CSCD 北大核心 2005年第11期685-688,共4页
目的研究ATP耗竭再恢复诱导的肾小管上皮细胞凋亡中,半胱氨酸天冬氨酸(caspase)3激活介导bcl-2降解的作用。方法使用代谢抑制剂短暂阻断细胞内ATP的生成,诱导细胞凋亡。以Hoechst 33342染色观察凋亡细胞。应用编码人bcl-2 RNA的腺病毒... 目的研究ATP耗竭再恢复诱导的肾小管上皮细胞凋亡中,半胱氨酸天冬氨酸(caspase)3激活介导bcl-2降解的作用。方法使用代谢抑制剂短暂阻断细胞内ATP的生成,诱导细胞凋亡。以Hoechst 33342染色观察凋亡细胞。应用编码人bcl-2 RNA的腺病毒感染细胞,使bcl-2在细胞内高表达。应用流式细胞仪,分析细胞凋亡和坏死的发生情况。以Western印迹检测caspase 3的活化以及bcl-2和bcl-2的降解产物。体外实验观察caspase 3和caspase 3特异性抑制剂DEVD对bcl-2降解产物的影响。结果高表达bcl-2组与对照组比较,细胞ATP耗竭60 min、90 min、120 min和再恢复60 min后,可减少细胞凋亡50%,P<0.05。肾小管上皮细胞ATP耗竭时,caspase 3被激活,bcl-2出现降解;细胞内ATP恢复时,bcl-2降解产物继续增多,并出现细胞凋亡。体外应用caspase 3可使bcl-2降解,而caspase3的特异性抑制剂DEVD能明显抑制caspase 3对bcl-2的降解。结论肾小管上皮细胞ATP耗竭再恢复时,caspase 3激活及其介导的bcl-2降解在细胞凋亡中发挥重要作用。 展开更多
关键词 半胱氨酸天冬氨酸3 基因 BCL-2 凋亡 肾小管 上皮细胞
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胆固醇栓塞综合征 被引量:1
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作者 Peter J Dupont Liz Lightstone +5 位作者 Elaine J Clutterbuck Gillian Gaskin Charles D Pusey Terry Cook Anthony N Warrens 赵元攸 《英国医学杂志中文版》 2004年第4期247-249,共3页
胆固醇栓塞是老年患者于动脉造影、血管外科手术、溶栓及抗凝治疗后常见的并发症,但这一疾病的诊断常被忽略。在此,我们介绍在过去6个月中发生于我们肾科病房的3个胆固醇栓塞的病例。每例均以急性肾衰为始发症状。
关键词 胆固醇栓塞综合征 并发症 肾病 诊断 治疗
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