老年2型糖尿病肾病患者尿液足细胞血清胱抑素C血清同型半胱氨酸水平与肾功能的相关性

目的 探讨老年2型糖尿病肾病患者尿液足细胞、血清胱抑素C（Cys-c）、血清同型半胱氨酸（Hcy）水平与肾功能的相关性。方法 选取2015年6月至2017年6月首都医科大学附属地坛医院收治的98例老年2型糖尿病肾病患者,纳入观察组。选取同期于我院行健康体检的98例老年健康志愿者为对照组。比较两组患者尿液足细胞、血清Cys C、Hcy水平与肾功能检测等相关生化指标的差异,采用Pearson相关性分析尿液足细胞、血清CysC、Hcy水平与肾功能指标的相关性。结果 观察组患者尿液足细胞、血清Cys C、Hcy水平均高于对照组,差异有统计学意义（P< 0.05）,肾功能指标：血肌酐（Cr）、血清尿素（BUN）、血清β2微球蛋白（β2-MG）、尿酸（UA）均高于对照组患者,差异有统计学意义（P<0.05）。Pearson相关性分析显示老年糖尿病患者尿液足细胞、血清Cys C、Hcy水平与患者肾功能指标（Cr、BUN、β2-MG、UA）均呈正相关（r>0,P<0.05）。结论 老年2型糖尿病患者尿液足细胞、血清Cys C、Hcy水平与肾功能呈正相关。     

PLCE1基因突变与激素耐药性肾病综合征的关系

磷脂酶Cε1(phospholipase C epsilon-1，PLCE1) 基因定位于常染色体10q23.32-q24.1,其编码蛋白为磷脂酶Cε1(phospholipase C epsilon-1, PLCε1)。PLCε1蛋白是最新发现的一种磷脂酶C(phospholipase C,PLC)同工酶，在成熟的肾小球足细胞表达,参与肾小球毛细血管袢的形成和正常发育。近来发现PLCE1 基因突变可以引起常染色体隐性遗传性肾病综合征。PLCE1基因移码或无义突变可导致婴儿早发性肾病综合征，其病理特征为弥漫性肾小球硬化(diffuse mesangial sclerosis，DMS)；而错义突变可导致局灶性节段性肾小球硬化(focal segmental glomerulosclerosis，FSGS)。但其确切发病情况及机制目前尚不清楚，本文就PLCE1基因及其编码蛋白与激素耐药性肾病综合征之间的关系作一简要综述。     

Redistribution of connexin43 expression in glomerular podocytes predicts poor renal prognosis in patients with type 2 diabetes and overt nephropathy.

BACKGROUND: Significance of podocyte injury in the progression of diabetic nephropathy is not well-understood. In this study, we examined whether alteration of gap junction protein connexin43 (Cx43) expression in podocytes is associated with the progression of overt diabetic nephropathy. METHODS: We recruited 29 type 2 diabetic patients with overt nephropathy who underwent renal biopsy. Nephrectomized kidney samples obtained from seven subjects with localized neoplasm and biopsy specimens from five patients diagnosed as minor glomerular abnormalities were used as controls. Cx43 staining on paraffin-embedded kidney sections were studied by immunohistochemistry. RESULTS: In controls, Cx43 was expressed at podocytes in a linear pattern along the glomerular basement membrane. In contrast, downregulation and loss of uniformly linear staining of Cx43 (Cx43 heterogeneity) in podocytes were observed in diabetic nephropathy. Cx43 intensity correlated with current renal function (R = 0.647, P < 0.005), whereas the magnitude of Cx43 heterogeneity correlated well with the degree of future decline in renal function (R = -0.705, P < 0.001). CONCLUSIONS: Alteration of Cx43 expression in podocytes was closely associated with the progression of overt diabetic nephropathy. These results indicate that change in Cx43 expression at podocytes represents a progressive stage in overt diabetic nephropathy and that it may be a convenient way to predict future decline in renal function.     

Roscovitine在衣霉素诱导足细胞损伤中的保护作用

目的 应用衣霉素诱导内质网应激（ERS）, 观察 Roscovtine 对 ERS 导致足细胞损伤的保护作用。方法体外培养永生化小鼠足细胞, 取 37 ℃分化成熟细胞随机分组：（1）正常对照组、 DMSO 组及衣霉素 Tunicamycin（1.0 μmol/L, TM） 组, 各实验组分别刺激 3、 6、 12 h。（2） 正常对照组、 Tunicamycin （1.0 μmol/L, TM） 组及 Tunicamycin+Rosco⁃ vitine（20、 40 μmol/L, TM+ROS）组, 各实验组分别刺激 12 h。应用流式细胞术及 TUNEL 法检测足细胞凋亡情况; 应用 Western blot 检测细胞周期素依赖性蛋白激酶 5（Cdk5）及 ERS 标志蛋白 GRP78、 Caspase-12、 CHOP 的表达变化。结果 （1）与正常对照组和 DMSO 组相比, Tunicamycin 刺激 3、 6 及 12 h 后, TM 组足细胞凋亡率及 Cdk5、 GRP78、 Caspase-12 和 CHOP 蛋白的表达水平均呈明显的时间依赖性增高 （P < 0.05）;（2） 加入 Roscovitine 干预后, 与 TM 组相比, TM+ROS（20、 40 μmol/L）组足细胞凋亡率及 GRP78、 Caspase-12 和 CHOP 蛋白的表达水平均显著降低（P < 0.05）, 其干预作用呈现明显的剂量依赖性。结论 Cdk5 抑制剂 Roscovitine 能显著抑制衣霉素诱导的足细胞凋亡,从而发挥细胞保护作用。Roscovitine 的足细胞保护作用可能有助于糖尿病肾病的治疗。     

REST and Stress Resistance in the Aging Kidney

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Serine Protease HTRA1 as a Novel Target Antigen in Primary Membranous Nephropathy

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Autonomous Calcium Signaling in Human and Zebrafish Podocytes Controls Kidney Filtration Barrier Morphogenesis

BackgroundPodocytes are critical to maintaining the glomerular filtration barrier, and mutations in nephrotic syndrome genes are known to affect podocyte calcium signaling. However, the role of calcium signaling during podocyte development remains unknown.MethodsWe undertook live imaging of calcium signaling in developing podocytes, using zebrafish larvae and human kidney organoids. To evaluate calcium signaling during development and in response to channel blockers and genetic defects, the calcium biosensor GCaMP6s was expressed in zebrafish podocytes. We used electron microscopy to evaluate filtration barrier formation in zebrafish, and Fluo-4 to detect calcium signals in differentiating podocytes in human kidney organoids.ResultsImmature zebrafish podocytes (2.5 days postfertilization) generated calcium transients that correlated with interactions with forming glomerular capillaries. Calcium transients persisted until 4 days postfertilization, and were absent after glomerular barrier formation was complete. We detected similar calcium transients in maturing human organoid glomeruli, suggesting a conserved mechanism. In both models, inhibitors of SERCA or IP3 receptor calcium-release channels blocked calcium transients in podocytes, whereas lanthanum was ineffective, indicating the calcium source is from intracellular podocyte endoplasmic-reticulum stores. Calcium transients were not affected by blocking heartbeat or by blocking development of endothelium or endoderm, and they persisted in isolated glomeruli, suggesting podocyte-autonomous calcium release. Inhibition of expression of phospholipase C-γ1, but not nephrin or phospholipase C-ε1, led to significantly decreased calcium activity. Finally, blocking calcium release affected glomerular shape and podocyte foot process formation, supporting the critical role of calcium signaling in glomerular morphogenesis.ConclusionsThese findings establish podocyte cell–autonomous calcium signaling as a prominent and evolutionarily conserved feature of podocyte differentiation and demonstrate its requirement for podocyte foot process formation.     

A Rare Autosomal Dominant Variant in Regulator of Calcineurin Type 1 (RCAN1) Gene Confers Enhanced Calcineurin Activity and May Cause FSGS

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温阳活血利水方含药血清对足细胞组织蛋白酶L、发动蛋白以及微丝、微管的影响

目的探讨温阳活血利水方治疗原发性肾病综合征蛋白尿的可能作用机制。方法将足细胞分为正常对照组、模型组、地塞米松组、10%中药血清组、20%中药血清组、空白对照组。除正常对照组、空白对照组外,其余各组用氨基核苷嘌呤霉素粉剂制备足细胞损伤模型。正常对照组和模型组加入完全培养基5 ml,地塞米松组加入392μg/L地塞米松5 ml,10%中药血清组、20%中药血清组、空白对照组分别加入10%中药血清、20%中药血清、空白对照血清各5 ml。RT-PCR法与Western blot分别检测各组足细胞胞浆组织蛋白酶L(CatL)、发动蛋白mRNA和蛋白表达含量,并用激光共聚焦观察各组中足细胞骨架微丝、微管的形态结构变化。结果与正常组比较,模型组CatL mRNA与蛋白含量升高,发动蛋白表达含量降低(P<0.01);与模型组比较,各中药血清组与地塞米松组CatL mRNA与蛋白表达含量降低,足细胞发动蛋白蛋白表达含量升高(P<0.01);与10%中药血清组比较,20%中药血清组CatL mRNA与蛋白表达含量降低,发动蛋白表达含量升高(P<0.01)。各组间足细胞发动蛋白mRNA表达含量差异无统计学意义(P>0.05)。结论温阳活血利水方可能通过下调足细胞CatL的表达,减少CatL对发动蛋白的酶解,从而稳定细胞骨架中的微丝、微管等结构,减少足突融合,改善蛋白尿。