足细胞抗原在人类膜性肾病治疗中的研究进展

膜性肾病(membranous nephropathy,MN)是一种器官特异性自身免疫病,发病机制是自身抗体结合足细胞靶抗原后激活补体导致肾小球滤过屏障损伤和蛋白尿.近年研究已发现中性肽链内切酶、M型磷脂酶A2受体(phospholipase A2 receptor,PLA2R)、醛糖还原酶、超氧化物歧化酶2、α烯醇化酶、1型血小板反应蛋白7A域等足细胞靶抗原.抗PLA2R抗体和肾组织PLA2R抗原是诊断和治疗MN的新兴生物标志物.     

PTEN在糖尿病肾病患者足细胞损伤中的作用

目的：探讨磷酸酰肌醇3激酶/蛋白激酶B( phosphoinositide 3 kinese/protein kinase B, PI3K/Akt)信号通路抑制因子PTEN与糖尿病肾病( diabetic nephropathy, DN)患者足细胞损伤的关系。方法收集30例DN患者和10例健康志愿者的24 h尿标本,采用酶联免疫吸附实验( enzyme linked immunosorbent assay, ELISA)检测患者尿液中足盂蛋白( podocalyxin, PCX)的水平;肾活检组织进行形态学观察,根据肾小球病变将DN患者分为3组,免疫组化法检测各组肾小球内p-Akt和PTEN的表达。结果 DN患者尿液中PCX的水平明显高于健康对照组,并随肾小球病变加重其水平逐渐上升;p-Akt和PTEN在DN患者肾小球的表达有所上调,但随着肾小球病变的加重其表达逐渐减少;DN患者尿液中PCX水平与PTEN表达呈负相关,与24 h尿蛋白呈正相关。结论 PTEN表达下调可能通过改变Akt的活化状态,从而在DN患者足细胞损伤中发挥一定作用。     

过表达APMAP减轻阿霉素肾病肾小球足细胞损伤

目的 探讨脂肪细胞膜相关蛋白质(APMAP)过表达对阿霉素(ADR)肾病肾小球足细胞损伤的影响。方法 采用尾静脉注射ADR构建阿霉素肾病大鼠模型,免疫组化观察肾组织中APMAP、NF-κB p65蛋白表达情况。构建APMAP基因过表达的鼠源肾小球足细胞MPC-5细胞株,并以0.5μmol/L ADR体外诱导构建足细胞损伤模型,再联合NF-κB信号通路激活剂CU-T12-9进行处理。CCK-8检测细胞增殖活性;ELISA检测乳酸脱氢酶(LDH)活性;流式细胞术检测细胞凋亡率;Western blot检测NF-κB p65、p-NF-κB p65、TNF-α等蛋白表达。结果 APMAP在阿霉素肾病大鼠肾组织中低表达,而NF-κB p65高表达(P<0.05)。APMAP过表达可提高ADR暴露下MPC-5细胞增殖活性,降低LDH活性及细胞凋亡率,下调NF-κB p65、p-NF-κB p65、TNF-α等蛋白表达(P<0.05);联合CU-T12-9处理可显著抑制APMAP过表达对ADR暴露下MPC-5细胞损伤的改善作用。结论 过表达APMAP可抑制ADR诱导的肾小球足细胞损伤,...     

IgA肾病合并膜性肾病1例

患者男性,52岁,因发现小便起泡沫半年,无水肿及其他不适入院。尿常规：潜血（＋＋）,蛋白（＋＋＋）,院外按“慢性肾炎”治疗1个月,尿蛋白无减少。体检：BP120/75mmHg,下肢无水肿,尿常规：蛋白（＋＋＋）,潜血（＋＋＋）,镜检可铜陵大量颗粒管型,24h尿蛋白排泄量2.95g,尿沉渣;红细胞123个μl,变形率85%,芽孢状;     

VEGF受体抑制剂对Ⅰ型糖尿病肾病大鼠足细胞病的治疗作用

目的:探讨血管内皮生长因子(VEGF)受体抑制剂SU5416对糖尿病肾病足细胞病的治疗作用.方法:SPF级SD雄性大鼠共30只,设为正常对照组(NC)、糖尿病肾病模型组(DN)、SU5416治疗组(SU5416),每组10只.DN大鼠由链脲菌素(STZ)诱导形成.SU5416治疗后第8周,分别测定大鼠体重( body weight,BW)、肾重(kidney weight,KW)、血糖( glucose,Glu)、大鼠24h尿蛋白排泄率(24 h UAER)和血肌酐(Scr).免疫荧光技术观察肾脏足细胞标记蛋白nephrin和podocin表达,Real time-PCR技术检测nephrin、podocin和VEGF mRNA水平,并观察肾脏病理变化.结果:与NC组大鼠相比,DN组大鼠BW明显下降,KW增加显著(P＜0.05),24 h UAER、Glu和Scr均明显升高,肾组织VEGF mRNA水平明显升高,nephrin、podocin蛋白水平及mRNA水平明显下降(P＜0.05),肾小球基底膜增厚、系膜基质增生;SU5416治疗后大鼠肾脏病理改善,KW较DN组明显下降,24hUAER明显降低,nephrin、podocin蛋白水平及mRNA水平上调(P＜0.05),但BW、Glu、Scr和VEGF mRNA水平治疗前后无统计学差异(P＞0.05).结论:SU5416治疗能降低DN大鼠24 h UAER、改善肾脏病理表现和上调肾小球足细胞标记蛋白nephrin和podocin水平.表明VEGF受体抑制剂对DN的治疗作用与其足细胞保护有关,抑制VEGF活性可能成为控制DN足细胞病进展的治疗手段之一.     

VEGF受体抑制剂对I型糖尿病肾病大鼠足细胞病的治疗作用

目的:探讨血管内皮生长因子(VEGF)受体抑制剂SU5416对糖尿病肾病足细胞病的治疗作用。方法:SPF级SD雄性大鼠共30只,设为正常对照组(NC)、糖尿病肾病模型组(DN)、SU5416治疗组(SU5416),每组10只。DN大鼠由链脲菌素(STZ)诱导形成。SU5416治疗后第8周,分别测定大鼠体重(body we ight,BW)、肾重(k idney we ight,KW)、血糖(glucose,G lu)、大鼠24 h尿蛋白排泄率(24 h UAER)和血肌酐(Scr)。免疫荧光技术观察肾脏足细胞标记蛋白nephrin和podoc in表达,Real tim e-PCR技术检测nephrin、podoc in和VEGF mRNA水平,并观察肾脏病理变化。结果:与NC组大鼠相比,DN组大鼠BW明显下降,KW增加显著(P<0.05),24 h UAER、G lu和Scr均明显升高,肾组织VEGF mRNA水平明显升高,nephrin、podoc in蛋白水平及mRNA水平明显下降(P<0.05),肾小球基底膜增厚、系膜基质增生;SU5416治疗后大鼠肾脏病理改善,KW较DN组明显下降,24 hUAER明显降低,nephrin、podoc in蛋白水平及mRNA水平上调(P<0.05),但BW、G lu、Scr和VEGF mRNA水平治疗前后无统计学差异(P>0.05)。结论:SU5416治疗能降低DN大鼠24 h UAER、改善肾脏病理表现和上调肾小球足细胞标记蛋白nephrin和podoc in水平。表明VEGF受体抑制剂对DN的治疗作用与其足细胞保护有关,抑制VEGF活性可能成为控制DN足细胞病进展的治疗手段之一。     

厄贝沙坦对早期糖尿病肾病大鼠足细胞nephrin表达的影响

目的： 观察nephrin在糖尿病肾病（DN）大鼠肾小球足细胞中的表达，探讨厄贝沙坦对DN肾脏保护作用的机制。方法： 应用链脲佐菌素建立大鼠DN模型，将DN模型大鼠随机分为2组：厄贝沙坦治疗组、模型对照组；另设正常对照组。各组分别干预8周后，观察大鼠体重、肾重、相对肾重、24 h尿蛋白定量、血糖、血清尿素氮、肌酐、总胆固醇、甘油三酯变化，利用光镜、电镜观察肾脏病理改变，应用免疫组化技术观察nephrin表达情况。结果： 应用厄贝沙坦干预后，DN大鼠24 h尿蛋白定量明显减少、肾功能改善、肾脏病理改变显著减轻、足细胞nephrin表达量明显减少。结论： 厄贝沙坦对DN肾脏的保护作用可能与其抑制足细胞nephrin表达有关。     

Sos2抑制NFATc1介导的糖尿病肾病足细胞损伤的机制研究

目的:观察高糖刺激对体外培养的小鼠肾足细胞鸟苷酸交换因子Sos2(Son of Sevenless homolog2)表达的影响,并初步探讨Sos2在高糖诱导足细胞损伤中的作用及其可能的分子机制。方法:通过免疫荧光染色及激光共聚焦显微镜观察Sos2在糖尿病肾病患者足细胞中的表达;体外培养小鼠永生化足细胞,以高糖(30 mmol/L葡萄糖)刺激足细胞48 h,采用RT-PCR、Western blot和免疫荧光检测高糖刺激下足细胞Sos2的mRNA及蛋白表达;采用Western bolt实验、免疫荧光及划痕实验检测Sos2过表达及沉默后podocin的表达、足细胞的活动性及NFATc1的入核情况;并采用RT-PCR检测NFATc1下游目的基因转录情况。结果:Sos2在糖尿病肾病患者的足细胞及高糖刺激体外培养的足细胞中表达显著降低(P 0. 05);沉默Sos2后,足细胞标志蛋白podocin表达显著降低,足细胞活动性增加,NFATc1入核增加,NFATc1下游目的基因转录增加(P 0. 05);与之相反,过表达Sos2组podocin表达显著增高,足细胞的活动性降低,NFATc1的入核减少,NFATc1下游目的基因转录降低(P 0. 05)。结论:Sos2可能通过抑制NFATc1入核而减轻糖尿病肾病足细胞损伤。     

UCH-L1在高糖刺激的足细胞及大鼠糖尿病肾病模型中的表达

目的探讨泛素羧基末端水解酶-1(ubiquitin carboxy-terminal hydrolase-1,UCH-L1)在高糖诱导的体外足细胞及糖尿病肾病大鼠模型体内足细胞中的表达。方法以25 mmol/L高糖刺激体外足细胞0、12、24 h后,提取细胞蛋白,采用Western bolt法检测UCH-L1的表达。20只8~10周龄Wistar大鼠,随机分为对照组和糖尿病肾病模型(STZ)组并造模,各10只,定期检测体重、血糖变化,处死大鼠取肾脏组织,HE染色和PAS染色观察肾脏形态学改变,采用免疫组化PV 9000两步法染色以WT1定位足细胞,检测足细胞内UCH-L1的表达。结果在不同时间高糖处理后,足细胞UCH-L1蛋白表达量随时间延长明显升高(P0.01);体重、血糖检测及HE、PAS染色证明糖尿病肾病模型构建成功,STZ组足细胞(WT1定位)内UCH-L1呈阳性,而对照组几乎无明显表达。结论分子生物学和形态学两种方法联合证明高糖可诱导体内外足细胞中UCH-L1的表达升高。     

IgA肾病足突融合和疾病严重性

目的　 Ig A肾病是一种与肾小球系膜细胞增生及 Ig A沉积有关的常见的进展性肾小球疾病。当前研究的设计为调查 Ig A肾病患者病变严重程度在功能上与形态上一致性改变。方法　通过观察 17例成年 Ig A肾病患者菊粉糖、对氨基马尿酸 (PAH)、 3H-非科尔的清除率和形态测定方法来研究肾小球的血液动力学、选择性滤过及超微结构。大分子物质通过半透膜渗透的数学模型用作特征性的肾小球选择性滤过。对照为 14名健康活体供肾者和 12名健康志愿者。结果　患者在病变严重程度上存在差异 ,但作为肾小球滤过率 (GFR)降低和蛋白尿排泄增加的一组…     

Podocyte loss involves MDM2‐driven mitotic catastrophe

Podocyte apoptosis as a pathway of podocyte loss is often suspected but rarely detected. To study podocyte apoptosis versus inflammatory forms of podocyte death in vivo, we targeted murine double minute (MDM)‐2 for three reasons. First, MDM2 inhibits p53‐dependent apoptosis; second, MDM2 facilitates NF‐κB signalling; and third, podocytes show strong MDM2 expression. We hypothesized that blocking MDM2 during glomerular injury may trigger p53‐mediated podocyte apoptosis, proteinuria, and glomerulosclerosis. Unexpectedly, MDM2 blockade in early adriamycin nephropathy of Balb/c mice had the opposite effect and reduced intra‐renal cytokine and chemokine expression, glomerular macrophage and T‐cell counts, and plasma creatinine and blood urea nitrogen levels. In cultured podocytes exposed to adriamycin, MDM2 blockade did not trigger podocyte death but induced G2/M arrest to prevent aberrant nuclear divisions and detachment of dying aneuploid podocytes, a feature of mitotic catastrophe in vitro and in vivo. Consistent with these observations, 12 of 164 consecutive human renal biopsies revealed features of podocyte mitotic catastrophe but only in glomerular disorders with proteinuria. Furthermore, delayed MDM2 blockade reduced plasma creatinine levels, blood urea nitrogen, tubular atrophy, interstitial leukocyte numbers, and cytokine expression as well as interstitial fibrosis. Together, MDM2‐mediated mitotic catastrophe is a previously unrecognized variant of podocyte loss where MDM2 forces podocytes to complete the cell cycle, which in the absence of cytokinesis leads to podocyte aneuploidy, mitotic catastrophe, and loss by detachment. MDM2 blockade with nutlin‐3a could be a novel therapeutic strategy to prevent renal inflammation, podocyte loss, glomerulosclerosis, proteinuria, and progressive kidney disease. Copyright © 2013 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.     

富马酸二甲酯对糖尿病肾病自噬及足细胞功能的影响

目的 探究富马酸二甲酯(DMF)对糖尿病肾病(DN)自噬及足细胞功能的影响.方法 通过对大鼠腹腔注射50 mg/kg的STZ诱导了DN模型,DN组大鼠灌胃2 mL生理盐水,DN+25DMF组、DN+50DMF组和DN+75DMF组分别灌胃25、50和75 mg/(kg?d)的DMF,共治疗12周.检测各组大鼠的血糖、尿...     

Correlation of Proteinuria with Podocyte Foot Process Effacement in IgA Nephropathy: An Ultrastructural Study

Background: Proteinuria is an uncommon clinical manifestation of IgA nephropathy and is usually seen in cases with severe lesions like endocapillary proliferation. However, it is occasionally seen even with cases with mild glomerular manifestations and may even be of nephrotic range.

Predictor: Podocyte foot process effacement.

Outcome: Severity of proteinuria.

Measurements: Podocyte foot process effacement was measured. Morphometric analysis was performed on transmission electron microscope images using a computerized digital photomicrograph system (BioWizard 4.2 Image analysis software, New Delhi, India). Proteinuria was measured quantitatively assigned into five grades.

Results: It was found that as the extent of proteinuria increased, the effacement ratio also increased, and this was most significant between “no” proteinuria and the rest of the categories.

Conclusion: Nephrotic presentation in IgA nephropathy is a known phenomenon and in certain cases may show near normal glomerular morphology with severe foot process effacement on EM being the only significant finding to explain the proteinuria. Proteinuria in these cases shows a significant correlation with degree of foot process effacement. Renal biopsy is important in these cases because they are known to have a better prognosis and are usually steroid responsive.     

Mesenchymal Stem Cells Ameliorate Podocyte Injury and Proteinuria in a Type 1 Diabetic Nephropathy Rat Model

Mesenchymal stem cells (MSC) attenuate albuminuria and preserve normal renal histology in diabetic mice. However, the effects of MSC on glomerular podocyte injury remain uncertain. The aim of this study was to evaluate the effects of MSC on podocyte injury in streptozotocin (STZ)-induced diabetic rats. Thirty days after diabetes induction by STZ injection (65 mg/kg, intraperitoneally) in Sprague-Dawley rats, the diabetic rats received medium or 2 × 10⁶ enhanced green fluorescent protein-labeled MSC via the renal artery. In vivo tracking of MSC was followed by immunofluorescence analysis. Diabetes-related physical and biochemical parameters were measured on day 60 after the MSC infusion. The expression of podocyte markers (nephrin and podocin), podocyte survival factors (VEGF and BMP-7), and the ultrastructural pathology of podocytes were also assessed. MSC were only detected in the glomeruli from the left kidney receiving MSC infusion. Compared with medium-treated diabetic rats, rats treated with MSC showed a suppressed increase in kidney weight, kidney to body weight index, creatinine clearance rate, and urinary albumin to creatinine ratio; however, the treatment had no effect on blood glucose or body weight levels. Furthermore, the MSC treatment reduced the loss of podocytes, effacement of foot processes, widening of foot processes, thickening of glomerular basal membrane (GBM), and loss of glomerular nephrin and podocin. Most important, MSC-injected kidneys expressed higher levels of BMP-7 but not of VEGF. Our results clearly demonstrated that intra-arterial administration of MSC prevented the development of albuminuria as well as any damage to or loss of podocytes, though there was no improvement in blood sugar levels. The protective effects of MSC may be mediated in part by increasing BMP-7 secretion.     

糖尿病肾病的血液流变学改变

本文对21例糖尿病肾病、14例糖尿病非肾病以及16例正常对照的血液流变学检查结果进行了分析.结果表明,两组糖尿病患者均呈现高粘滞血症,尤以糖尿病肾病组为著.糖尿病肾病组患者尿蛋白排泄量与血浆比粘度、红细胞刚性指数、血小板粘附率呈正相关.内生肌酐清除率(Ccr)与上述三项指标呈负相关.     

Smoking-Related Renal Histologic Injury in IgA Nephropathy Patients

Purpose

Smoking reportedly exerts deleterious effects on renal function; however, its effects on histology have not been clarified in patients with IgA nephropathy (IgAN).

Materials and Methods

Renal histology was evaluated in a cohort of 397 patients diagnosed with IgAN according to smoking status and dose in relation to renal function.

Results

Among the study cohort, which was predominantly male (88.5%), 52 patients (13%) were current smokers. These current smokers demonstrated more frequent hypertension and higher serum creatinine levels than non/ex-smokers at the time of diagnosis, which was apparent with increased smoking dose. The percentages of global glomerulosclerosis and arteriolar hyalinosis increased with increased smoking dose, whereas tubulointerstitial fibrosis or arterial intimal thickening did not. Glomerular mesangial alpha-smooth muscle actin expression were similar between current and non/ex-smokers matched for age, gender, hypertension, and histologic severity, although the number of glomerular CD68+ cells was significantly fewer in smokers. Initial serum creatinine level, estimated glomerular filtration rate (eGFR), and global glomerulosclerosis were found to be risk factors of serum creatinine doubling in both smokers and non/ex-smokers by univariate analysis during a mean follow-up of 3.8 years.

Conclusion

In addition to dose dependent renal functional decline and hypertension, smoking contributes to renal disease progression by eliciting microvascular injury in IgAN patients.     

C肽在糖尿病肾病中的临床意义

目的 了解糖尿病肾病患者C肽分泌特点及临床意义.方法 ①选取行胰岛功能检查的住院患者86例,依出院诊断结果,将其分为两组:仅糖尿病组(DM)、糖尿病肾病组(DN),并将DN分为5期(Ⅰ～Ⅴ期);②比较两组相关因素有无差异并分析DN患者C肽分泌特点;③分析DN分期特点及各期C肽分泌特点.结果 ①DN组和仅DM组相比,性别、年龄、血压、血脂、空腹血糖均无统计学差异(P＞0.05);②两组C肽分泌高峰均延迟,DN组分泌高峰在餐后3h,而仅DM组在餐后2小时;③DN以Ⅲ、Ⅳ期多见,占DN的91.9％,相比而言,Ⅰ、Ⅱ期较少,仅占1.6％;任一期的C肽分泌高峰均在餐后3h;对比各期的C肽值,差异无统计学意义(P＞0.05).结论 DN患者的相关因素与仅DM患者相似,当发现DN时,患者肾脏及胰岛功能往往受损较重,但C肽与DN不同分期间无明显相关性.     

糖尿病肾病患者血红蛋白浓度的研究

目的探讨糖尿病肾病患者血红蛋白浓度与终末期肾病之间的关系,及血红蛋白浓度与其它临床指标之间的关系。方法回顾性分析203例糖尿病肾病患者的临床资料和实验室资料;根据血红蛋白浓度的不同将研究对象分为三组,A组Hb〈100g/L,B组100g/L≤Hb〈120g/L,C组Hb≥120g/L。结果三组血肌酐的值分别为（432.74±37.60）μmol/L、（220.32±23.88）μmol/L、（111.67±10.73）μmol/L,各组间差异具有统计学意义（P〈0.05）。血红蛋白浓度越低血肌酐值越高,到达终末期肾病患者的比例越高。各组收缩压、尿素氮、尿酸的组间差异也具有统计学意义（P均〈0.05）。结论低血红蛋白浓度是终末期肾病的危险因素;血红蛋白浓度对其它一些临床指标的数值有一定的影响。