雷公藤多苷对糖尿病肾病大鼠足细胞病变的影响

目的 探讨雷公藤多苷（TWP）对糖尿病肾病大鼠足细胞病变的影响。 方法 SD大鼠100只,按随机数字表法分为正常对照组（A组）、糖尿病组（B组）与TWP治疗组（C组）。并将C组按4、8、16 mg?kg-1?d-1不同给药剂量分为3组（Ca、Cb、Cc）。糖尿病组与TWP治疗组大鼠分别给予链脲菌素（STZ）一次性腹腔注射建立糖尿病大鼠模型。12周后检测24 h尿蛋白量、血肌酐（Scr）、尿素氮（BUN）、外周血白细胞（WBC）、血糖（Glu）、肾质量/体质量（KW/BW）的变化;HE染色检测肾组织病理变化;透射电镜测量足细胞超微结构变化;免疫荧光法检测肾皮质nephrin和podocin蛋白表达。 结果 （1）与A组比较,B组及C组Scr、BUN增高（P ＜ 0.05）;Glu、KW/BW和24 h尿蛋白量显著增高（P ＜ 0.01）,除Cc组（3/20）出现肝酶（ALT、AST）增高及WBC下降外,各组ALT、AST、WBC差异无统计学意义;肾脏皮质nephrin和podocin蛋白表达减少,肾小球、小管间质及足细胞病变明显。（2）与B组比较,C组KW/BW和24 h尿蛋白量降低（P ＜ 0.01）,肾脏皮质nephrin和 podocin蛋白表达增高（均P ＜ 0.01）,肾小球、小管间质及足细胞病变明显减轻,并随雷公藤多苷剂量的增加而越加明显。 结论 TWP对糖尿病大鼠足细胞病变具有保护作用,并与剂量相关。其部分机制可能与上调nephrin和podocin蛋白表达有关。     

氟伐他汀对糖尿病肾病大鼠足细胞分布及排泄的影响

目的探讨氟伐他汀对DN大鼠足细胞分布及排泄的影响。方法将大鼠分为3组：对照组、DN模型组、氟伐他汀治疗组。腹腔注射链脲菌素（STZ）诱导DN大鼠模型。实验10周末测24小时尿蛋白定量（TP）、血清总胆固醇（TC）,间接免疫荧光法检测尿沉渣足细胞特异性标志蛋白podocalyxin（PCX）以检测尿液足细胞（UPC）水平;免疫荧光染色观察肾小球上皮细胞蛋白-1（GLEPP1）的分布。结果DN模型组UPC、TP、TC较对照组均明显升高;氟伐他汀治疗组TC、UPC及TP较DN模型组均降低;对照组GLEPP1正常、DN模型组呈节段性明显缺失、氟伐他汀治疗组缺失较轻。UPC与TP呈正相关,与TC无显著相关性。结论尿液中脱落足细胞检测可作为判断DN病情活动性的标志之一。氟伐他汀可减轻DN大鼠尿蛋白、降低胆固醇、减少足细胞脱落及排泄。     

糖尿病肾病模型大鼠中存在蛋白降解途径失调

目的 研究蛋白酶体和自噬两条主要蛋白质降解途径在细胞体外及糖尿病肾病（DN）模型大鼠体内中的变化。 方法 原代培养SD大鼠肾小球系膜细胞,观察该细胞在高糖环境中的增殖情况。分别给予30 mmol/L葡萄糖（HG组）、5.4 mmol/L葡萄糖（NC组）孵育0 h、8 h、16 h、72 h,并用Western印迹法检测蛋白酶体途径蛋白酶体α各亚基（PSMAs）和自噬途径LC3蛋白的表达。以自发性2型DN模型OLETF大鼠和正常对照LETO大鼠作体内实验,每4周动态监测血糖、24 h尿蛋白量,于36周龄处死大鼠,取肾组织,观察病理形态学并进行半定量评分;利用Western印迹法分析肾皮质中PSMAs和LC3蛋白的表达。 结果 与NC组相比,高糖环境下8 h蛋白酶体α各亚基、LC3Ⅰ和LC3Ⅱ蛋白表达均降低,差异均有统计学意义（P < 0.05）。其他各时间点两组间差异无统计学意义。与正常对照LETO大鼠相比,OLETF大鼠血糖水平和24 h尿蛋白量逐渐升高,差异有统计学意义（P < 0.01）。OLETF大鼠肾小球硬化指数和肾小管间质纤维化指数均显著高于LETO大鼠,差异有统计学意义（P < 0.01）;OLETF大鼠36周龄肾皮质中PSMAs蛋白表达明显减少,与正常对照LETO大鼠差异有统计学意义（P < 0.05）,LC3Ⅱ蛋白表达也有降低趋势,但差异无统计学意义。 结论 蛋白降解途径失调可能在DN的发生、发展中起了一定的作用。     

Inhibition of cyclooxygenase-2 expression contributes to podocyte injury in streptozotocin-induced diabetic rats

Objective To investigate the role of cyclooxygenase-2 (COX-2) in podocyte injury in diabetic rats mediated by the disruption of low-density lipoprotein receptor (LDLr) pathway. Methods Eight-week old male Sprague-Dawley (SD) rats were treated for 12 weeks by dividing into three groups: control rats, streptozotocin (STZ) induced diabetic rats (DM), and diabetic rats treated with aspirin (DM+Aspirin). The plasma lipid profile was checked by clinical biochemistry assay. The ratio of urinary microalbumin to creatinine (ACR) was detected by enzyme-linked immunosorbent assay. Intracellular lipid accumulation was evaluated by Oil Red O staining and a free cholesterol quantitative assay. The glomerular podocyte injury and the expression of molecules related with LDLr pathway were evaluated by electron microscope, immunohistochemical staining, immunofluorescent staining, and Western blotting. Results There were increased levels of urinary ACR (P＜0.01) and podocyte injury(P＜0.01) in DM rats compared with the controls. Additionally, lipid accumulation in kidneys of DM rats were significantly increased (P＜0.01), due to increased protein expressions of COX-2, LDLr, sterol regulatory element–binding protein (SREBP) cleavage activating protein (SCAP), and SREBP-2 (P＜0.01). However, these changes were significantly inhibited by an inhibitor of COX-2, Aspirin (P＜0.05). It's worth noting that, COX-2 protein expression was closely correlated with LDLr protein expression (r=0.85, P＜0.01). Conclusion Dysregulation of LDLr pathway contributes to podocyte injury in diabetic nephropathy, which may be mediated through the increased COX-2 expression.     

活性维生素D通过调节糖尿病肾病大鼠巨噬细胞M1及M2表型活化防治足细胞损伤

目的 探讨活性维生素D（VD）能否通过调节肾组织巨噬细胞M1 及M2 表型活化从而防治糖尿病肾病（DN）大鼠足细胞损伤。 方法 利用腹腔注射链脲菌素（STZ）建立糖尿病大鼠模型。将SD 雄性大鼠按随机数字表法分为4 组：对照组1（NC-1 组,n=8）、对照组2（NC-2 组,n=8,对照+骨化三醇0.1 μg·kg^-1·d^-1 灌胃）、DN 组（n=24）、DN+VD 干预组（VD 组,n=24,DN+骨化三醇0.1 μg·kg^-1·d^-1 灌胃）,定期检测血糖、体质量,收集尿标本,分别于干预后8周、14周、18周末处死动物,检测Scr、BUN和尿蛋白变化;PAS染色观察肾脏病理改变;免疫组化法检测肾组织CD68+巨噬细胞浸润数量;Western 印迹法检测nephrin、podocin、CD68 以及M1巨噬细胞特异性标志物诱导型氮氧化物合酶（iNOS）、肿瘤坏死因子α（TNF-α）和M2巨噬细胞特异性标志物CD163、精氨酸酶1（Arg-1）、甘露糖受体（MR）表达。 结果 （1）与两对照组相比,DN 组Scr、BUN、24 h 尿蛋白量及肾小球系膜基质增生程度显著增加（P＜0.05）,podocin、nephrin蛋白表达下降（P＜0.05）。VD干预后能明显改善上述病理现象（均P＜0.05）。（2）与两对照组相比,DN组肾组织CD68⁺巨噬细胞浸润数量明显增加,呈时间依赖性。VD干预后能显著减少CD68⁺巨噬细胞浸润数量（P＜0.05）。（3）进一步确定肾组织巨噬细胞M1、M2活化表型发现,8周、14周、18周末DN组iNOS、TNF-α蛋白表达较对照组显著升高（均P＜0.05）,VD干预后能明显抑制同期DN肾组织iNOS、TNF-α表达（均P＜0.05）;8周、14周末VD组CD163、Arg-1、MR蛋白表达与DN组相比差异无统计学意义（均P＞0.05）,而18周末VD组CD163、Arg-1、MR蛋白表达较DN 组显著升高（均P＜0.05）,CD163/CD68 蛋白表达比例亦显著增加（P＜0.05）。（4）相关分析显示,M1 标志物iNOS 与nephrin、podocin 蛋白表达均呈负相关（r＝-0.707,P＜0.01;r＝-0.712,P＜0.01）;M2标志物CD163与nephrin、podocin蛋白表达均呈正相关（r＝0.627,P＜0.01;r＝0.613,P＜0.01）。 结论 活性维生素D具有调节巨噬细胞表型活化的能力,通过抑制巨噬细胞M1型活化并增强M2型活化,进而发挥足细胞保护作用。     

舒洛地特对糖尿病高血压大鼠足细胞podocalyxin表达的影响

目的 研究舒洛地特对糖尿病高血压大鼠的肾脏保护作用和足细胞podocalyxin（PCX）表达的影响。 方法 链脲佐菌素（STZ）注射后,醋酸脱氧皮质酮（DOCA）+盐建立糖尿病高血压大鼠模型。设对照组（CTL组）、模型组（STZ+DOCA组）、舒洛地特治疗组（GAG组）和舒洛地特+替米沙坦治疗组（GAG+ARB组）,每组各6只大鼠。于建模后0、2、4、6和8周测尾动脉压、尿白蛋白和8周末尿N-乙酰-β-葡萄糖苷酶（NAG）。8周末取血检测胰岛素、血肌酐（Scr）、胆固醇（TC）、三酰甘油 （TG）、Na+、K+。HE、PAS染色观察病理改变和肾小球正切面足细胞计数。免疫组织化学检测podocalyxin在肾小球表达和分布。RT-PCR和Western印迹法检测podocalyxin mRNA和蛋白表达。 结果 （1）GAG+ARB组4周末血压显著低于STZ+DOCA组和GAG组（P < 0.05）。GAG组和GAG+ARB组TG、TC和胰岛素与STZ+DOCA组差异无统计学意义。（2）6周末GAG+ARB组尿白蛋白量显著低于STZ+DOCA组 [（52.9±7.6） mg/24 h比（102.2±6.9） mg/24 h,P < 0.05];8周末GAG组和GAG+ARB组尿白蛋白量均显著低于STZ+DOCA组（P < 0.05）,而GAG+ARB组尿白蛋白量显著低于GAG组[（33.8±6.8） mg/24 h比(85.2±8.7) mg/24 h,P < 0.05]。GAG+ARB组和GAG组尿NAG均显著低于STZ+DOCA组（P < 0.05）。（3）GAG组和GAG+ARB组肾小球硬化指数（GSI）和间质纤维化指数（IF）均显著低于STZ+DOCA组（P < 0.05）,各组肾小球正切面足细胞数差异无统计学意义。（4）与STZ+DOCA组相比,GAG组PCX mRNA和蛋白表达显著增加,而GAG+ARB组PCX表达显著高于GAG组。 结论 舒洛地特可通过增加足细胞podocalyxin表达,减轻糖尿病高血压大鼠蛋白尿和病理损伤,与替米沙坦联用有叠加作用。     

乌索酸改善高糖诱导系膜细胞损伤的机制

目的 探讨乌索酸能否通过抑制高糖状态下系膜细胞内miRNA-21的过表达,上调其靶基因PTEN的表达,抑制磷脂酰肌醇-3-激酶(PI3K)-Akt-哺乳动物雷帕霉素靶蛋白(mTOR)信号通路的激活,诱导自噬,减少细胞外基质堆积,发挥其肾脏保护作用.方法 高糖培养大鼠肾小球系膜细胞,以PI3K抑制剂LY294002以及不同剂量乌索酸进行干预,应用甲基噻唑基四唑(MTT)法观察细胞增殖能力,总蛋白/总细胞数测定细胞肥大,Western印迹和实时定量PCR检测PTEN-PI3K-Akt-mTOR信号通路活性、Ⅰ型胶原及自噬标志物.透射电镜观察自噬体的形成.结果 与正常对照组相比,高糖培养的系膜细胞出现显著的肥大、增殖,细胞内miRNA-21表达明显上调,PTEN蛋白及mRNA的表达明显下调,p85PI3K、磷酸化(p)-Akt、p-mTOR、Ⅰ型胶原、p62/SQSTMI表达明显增加,LC3 II表达明显降低,差异均有统计学意义(均P＜ 0.01).与高糖组相比,乌索酸干预组及LY294002组细胞肥大、增殖程度均明显降低,p85PI3K、p-Akt、p-mTOR、Ⅰ型胶原、p62/SQSTMI的表达均明显降低,LC3 II表达明显升高,差异均有统计学意义(均P＜ 0.01);但LY294002组细胞内miRNA-21和PTEN的表达与高糖组差异无统计学意义,而乌索酸干预组细胞内miRNA-21的表达明显下调,PTEN的表达明显上调,差异均有统计学意义(均P＜ 0.01).结论 乌索酸可能通过抑制高糖培养系膜细胞内miRNA-21的过表达,上调PTEN表达,抑制PI3K-Akt-mTOR信号通路异常活化,增强自噬从而减少细胞外基质堆积,减轻细胞的肥大、增殖.     

二氢欧山芹通过调节自噬保护高糖诱导的足细胞损伤

目的 探讨二氢欧山芹(columbianetin,CBT)对高糖诱导的足细胞损伤保护作用及其机制研究.方法 体外培养永生化小鼠足细胞系MPC-5,给予高浓度的葡萄糖刺激,观察CBT对高糖诱导的足细胞损伤及自噬的影响;采用蛋白质印迹法检测足细胞自噬相关蛋白;荧光显微镜观察GFP-LC3-Ⅱ转染后足细胞绿色荧光颗粒.结果 ...     

他克莫司对阿霉素肾病大鼠足细胞损伤的修复作用

目的探讨他克莫司对阿霉素肾病大鼠足细胞损伤的修复作用。方法通过阿霉素尾静脉注射建立大鼠微小病变肾病模型,并以他克莫司进行干预。大鼠随机分为对照组、模型组和他克莫司组。每组分别于第0、7、14、21、28、35天采集尿液,检测尿蛋白排泄量,于造模第35天处死大鼠收集肾脏标本。电镜观察各组大鼠足细胞足突融合情况;TUNEL检测法对肾小球进行染色;免疫组化或免疫荧光对WT-1、caspase-3进行定位和半定量检测,观察大鼠足细胞的数量和凋亡。结果阿霉素尾静脉注射后大鼠尿蛋白显著增加,足细胞数目明显减少,足突广泛融合;与模型组比较,他克莫司组大鼠24 h尿蛋白明显减少,足突融合改善,足细胞数目增多。与对照组比较,模型组大鼠肾小球内caspase-3蛋白表达增加,TUNEL染色加深;与模型组比较,他克莫司组肾小球内caspase-3表达量下降,TUNEL阳性率减少,说明足细胞凋亡减少。结论他克莫司可修复阿霉素肾病大鼠足细胞损伤,其机制与抑制足细胞凋亡有关。     

The relationship between autophagy and renal tissue injury in adriamycin nephropathy rats

Objective To observe the formation of autophagosome, the expression and distribution of autophagy-related protein LC3-Ⅰ, LC3-Ⅱ and Beclin-1 in adriamycin nephropathy rats at different pathological periods, to explore the relationship between autophagy and renal tissue injury, the occurrence of proteinuria, the progression of renal disease. Methods Sixty normal male SD rats were randomly divided into control group (n=30) and model group (n=30), the rats in model group were injected with adriamycin(6.5 mg/kg) via tail - vein for one time, while the rats in control group were injected with saline. Urine protein quantitation of 24 hour, the levels of serum albumin and total cholesterol were measured serially at the 2, 4, 6, 8, 10 weeks. The changes of kidney tissue pathology were detected after HE, PAS and Masson staining by light microscope. The formation of autophagy were detected by transmission electron microscopy, the localization and distribution of LC3-Ⅰ, LC3-Ⅱ and Beclin - 1 were detected by indirect immunofluorescence staining in kidney tissue, the autophagy - related proteins LC3-Ⅰ, LC3-Ⅱ and Beclin-1 expression was detected by Western blotting. Results In model group, urinary protein began to increase at the first two weeks, serum albumin decreased at the same time, and total cholesterol increased in the four weeks. There was a statistically significant difference compared with the control group (P＜0.01). The Scr and BUN were increased slightly at the four weeks in model group, and showed the deterioration of renal function after the eight weeks. There was a statistically significant difference compared with the control group (P＜0.01). Mesangial cell proliferation, mitochondrial swelling and foot process broadening and integration appeared early in the model group, while foot process disappearing and nuclear pyknosis were observed in the late by transmission electron microscope; Renal pathology gradually changed from mesangial proliferation to focal segmental glomerulosclerosis (FSGS) by light microscope. A low expression of autophagy was detected in renal tissue of control group rats by transmission electron microscopy and immunofluorescence microscope; in model group, with the progression of disease, the autophagy was significantly enhanced and maintained at a high level. With the progression of disease, the autophagy- related proteins LC3-Ⅰ, LC3-Ⅱ and Beclin-1 was significantly enhanced in the model group than the control group (P＜0.05). Conclusion Autophagy is involved in renal tissue injury and the occurrence of proteinuria, closely related to the progression of renal disease.     

自噬在糖尿病肾病进展中的作用及机制

糖尿病肾病是导致终末期肾脏疾病的主要病因之一,其发病机制不清。自噬是一种高度保守的细胞学事件,能够降解细胞内异常蛋白和细胞器,维持细胞内环境稳定,在多种急慢性肾脏疾病中发挥着重要的作用。研究发现糖尿病肾病中肾脏自噬功能受损,提示自噬障碍可能参与糖尿病肾病发病。本文将针对肾脏不同类型细胞,对自噬在糖尿病肾病中作用的相关研究进展进行综述。     

Human umbilical cord mesenchymal stem cells co-culture ameliorates the innate immunity of podocytes induced by high glucose

Objective To explore the effects of human umbilical cord mesenchymal stem cells(HUC-MSCs) on the innate immunity of podocytes mediated by Toll-like receptor (TLR) signaling pathway under high glucose (HG) condition. Methods Podocytes were divided into four groups according to the treatment: normal glucose group (NG), mannitol control group (NG+MA), high glucose group (HG) and HUC-MSCs co-culture group (HG+MSC). After 72 hours treatment, the protein levels of interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), heat shock protein 70 (HSP70), high-mobility group box-1 (HMGB1) in culture medium were measured by ELISA. Real-time PCR was used to detect the mRNA expressions of TLR2 and TLR4. Western blotting was used to detect the protein expressions of TLR2, TLR4, myeloid differentiation factor 88 (MyD-88) and phospho-P65 (p-P65). Immunofluorescence staining was used to study the localization of p-P65 in podocytes. Results High glucose induced the inflammation of podocytes by activating the TLR signaling, which increased the secretion of IL-6, TNF-α, HSP70, HMGB1, the mRNA level of TLR2, TLR4 and the protein level of TLR2, TLR4, MyD-88 and p-P65 (all P＜0.05). High glucose also activated NF-κB and induced its nuclear translocation. HUC-MSCs co-culture decreased the inflammation and restrained the TLR signaling. Conclusions HUC-MSCs co-culture decreases the inflammation and innate immunity of podocytes induced by HG.     

糖肾平对糖尿病肾病大鼠足细胞nephrin与CD2AP蛋白及其mRNA表达的影响

目的：观察糖肾平对糖尿病肾病（DN）大鼠足细胞裂孔隔膜蛋白分子nephrin和CD2AP表达的影响,探讨其防治DN的作用机制.方法：雄性Wistar大鼠74只,按体重随机分组为正常组10只,造模组64只.造模组大鼠腹腔一次性注射链脲佐菌素（STZ）（45 mg/kg）,72 h后检测大鼠血糖和尿糖,以血糖≥16.7 mmol/L、尿糖（＋＋＋＋）者为DN大鼠造模成功.模型成功的大鼠按体重随机分为模型组、厄贝沙坦组、糖肾平小、大剂量组,各组分别干预14周.于第14周处死大鼠,无菌取肾,采用原位杂交和免疫组化技术观察肾组织足细胞裂孔隔膜蛋白nephrin和CD2AP蛋白及其mRNA的表达水平.结果：模型组与正常组相比,nephrin和CD2AP蛋白及mRNA表达水平明显下调（P〈0.01）;与模型组相比,厄贝沙坦、糖肾平干预后糖尿病大鼠肾小球足细胞nephrin和CD2AP蛋白及mRNA水平表达明显增加（P〈0.05）.结论：糖肾平可能通过维持足细胞裂孔隔膜蛋白nephrin和CD2AP的稳定性,从而发挥对DN肾脏的保护作用.     

Abated microRNA-21 attenuates high glucose-induced autophagy inhibition in rat mesangial cells by PTEN/Akt/mTOR pathway

Objective To investigate the effects of abated microRNA-21 (miRNA-21) on phosphatase and tensin homologue on chromosome ten protein (PTEN) and PI3K/Akt/mTOR pathway, as well as their further influence on the autophagy in high glucose (HG, 25.0 mmol/L) induced rat glomerular mesangial cells. Methods MiRNA-21 inhibitor and negative control were transfected by liposome 2000 into rat glomerular mesangial cells (HBZY-1). The cells were divided into normal glucose (5.5 mmol/L) group, normal glucose+negative control group, normal glucose+miRNA-21 inhibitor group, HG group, HG+negative control group and HG+miRNA-21 inhibitor group. Cell proliferation and hypertrophy were assayed by MTT and the ratio of total protein to cell number respectively. The miRNA-21 expression was detected using real time PCR. The expressions of PTEN/Akt/mTOR signaling signatures, autophagy-associated protein (p62 and LC3 Ⅱ) and collagen Ⅰ was detected by Western blotting and real time PCR. Autophagosomes were observed using electron microscopy. Results Compared with those in normal glucose group, in HG group cells had hypertrophy and proliferation, up-regulated miRNA-21 expression, and down-regulated PTEN protein and mRNA expressions (all P＜0.01). Also there were and up-regulated p-Akt, p-mTOR, p62 and collagen Ⅰ expression, and lower LC3 Ⅱ expression and autophagosomes (all P＜0.01). Further, compared with those in HG group, cells hypertrophy and proliferation in HG+miRNA-21 inhibitor group were reduced, expressions of p-Akt, p-mTOR, p62 and collagen Ⅰ were down-regulated, while expressions of PTEN and LC3 Ⅱ and autophagosomes were up-regulated (all P＜0.01). Conclusions MiRNA-21 inhibitor up-regulates PTEN expression, which inhibits the activation of Akt/mTOR signaling pathway, ameliorates cell hypertrophy, proliferation and enhances autophagy to reduce extracellular matrix accumulation.     

高糖对小鼠肾小球足细胞podocalyxin mRNA表达的影响

目的探讨高糖对小鼠肾小球足细胞podocalyxin mRNA表达的影响。方法以体外培养的小鼠足细胞为研究对象,将细胞分组,即正常对照组（NG组）：D-葡萄糖5．56mmol／L;高糖培养组（HG组）;D-葡萄糖25．0mmol／L。应用逆转录-PCR技术,检测高糖培养不同时间点对足细胞表达podocalyxin mRNA的影响;Western blot印迹分析检测细胞转录因子WT-1蛋白变化。结果①逆转录-PCR技术显示随着高糖培养时间增加,HG组肾小球足细胞与WG组比较、以及HG组（第6、12、24h）组间比较,podocalyxin mRNA表达均减少（P〈0．05）。②培养的足细胞表达基础水平的WT-1蛋白,但在经高糖培养2h后其表达即开始下降（P〈0．05）。结论高糖培养使小鼠肾小球足细胞podocalyxin mRNA和转录因子WT-1蛋白表达下降。     

Early autophagy activation inhibits podocytes from apoptosis induced by aldosterone

Objective To explore the protection of early autophagy activation on podocyte injury induced by aldosterone. Methods In vitro cultured mouse podocyte clones (MPC5) were treated with aldosterone for 6, 12, 24, 48 h respectively. Apoptosis of podocytes was detected by Annexin V combined with flow cytometry. After 24 h treatment with aldosterone, the existence of apoptotic body and autophagosome was observed by electron microscopy. The protein expressions of LC3, caspase?3 and nephrin were examined by Western blotting. The mRNA expression of Beclin?1 was detected by real?time PCR. Results The induction of apoptosis and autophagy by aldosterone in podocytes was in time?dependent mannner. After 24 h treatment with aldosterone, the apoptosis was increased by 26.5% (P＜0.05) and the expression of nephrin was decreased by 28.0% (P＜0.05) compared to control group. Aldosterone remarkably induced the expression of Beclin?1 at 6 h and promoted the transformation of LC3?Ⅰto LC3?Ⅱ at 12 h (P＜0.05). Compared to simple aldosterone treatment, the apoptosis rate of podocyte was increased by 39.0%（P＜0.05）and the expression of nephrin was declined by 19.5%（P＜0.05) after 3?methyladenine (3?MA) pre?treatment. Conclusions Aldosterone can induce autophagy and apoptosis in podocytes. Autophagy occurs earlier (12 h) than apoptosis (24 h). The occurrence of autophagy can inhibit the apoptosis,so the autophagy pathway may be a new research topic of glomerular disease treatment.     

2型糖尿病肾脏损害病理类型分类初探

目的 探讨2型糖尿病肾脏损害病理类型的分类方法。 方法 回顾性分析49例除外非糖尿病肾病的伴显性白蛋白尿2型糖尿病患者的肾脏病理表现及临床特点,根据病理表现分为典型糖尿病肾小球病（DG）组和不典型糖尿病相关肾脏疾病（ADRD）组。 结果 DG占59.2%,ADRD占40.8%。病理表现上,DG的肾小球系膜区体密度、肾小球基底膜厚度、肾小管间质病变积分和肾小动脉玻璃样变发生率均大于ADRD,而足细胞相对密度低于ADRD。临床表现上,DG的糖尿病病程较长,糖尿病视网膜病变（DR）发生率高,空腹血糖较高,收缩压和平均动脉压较高,尿蛋白量较多,GFR下降更明显,而ADRD的体质量指数和肥胖比例较高,血脂紊乱更显著。DG和ADRD的GFR均与肾小球球性硬化率呈负相关,而DG的尿蛋白量水平与肾小球系膜区体密度呈正相关,ADRD的尿蛋白量水平与病理指标无显著相关。对DG诊断预测价值较高的有DR（阴性预测值94.8%）和已知糖尿病病程超过5年（阴性预测值90.7%)。 结论 2型糖尿病肾损害的病理表现多样,ADRD与DG是两种差异显著的2型糖尿病肾损害的病理表现,区分ADRD与DG能更好地从临床预测病理。     

STZ诱导的糖尿病大鼠血清脂联素水平分析

目的 探讨STZ诱导的糖尿病大鼠血清脂联素浓度及其与糖尿病肾病的关系.方法 以链脲佐菌素诱导建立1型糖尿病大鼠模型,ELISA方法测定正常对照组(A组),1型糖尿病大鼠2周(B组),6周(C组),12周(D组)及治疗组(E组)的血清和尿脂联素浓度.结果 D组和C组较A组和B组血清和尿脂联素水平均升高(均P<0.01);D组较C组血清和尿脂联素水平升高(P<0.05).E组从6～12周经胰岛素治疗后,较D组血清和尿脂联素水平有所下降(P<0.05).在C组和D组血清脂联素与24h尿白蛋白排泄率,尿脂联素呈正相关(r=0.85,P<0.01;r=0.65,P<0.01).尿脂联素与尿白蛋白排泄率呈显著正相关(r=0.78,P<0.01).逐步回归分析提示血清脂联素受尿白蛋白排泄率影响最大(β=0.89,P<0.01).结论 在糖尿病肾病发生发展中,血清脂联素水平随尿白蛋白排泄率增加而升高,补充胰岛素后血清脂联素水平下降,因此,血清脂联素水平升高是机体的一种保护机制.     

Changes of autophagosomes of podocytes in idiopathic membranous nephropathy and its clinical significance

Objective To observe the quantity change of autophagosomes in podocytes and expressions of autophagy-related gene Beclin-1 and microtubule-associated protein 1 light chain 3 (LC3) in different pathological stages of idiopathic membranous nephropathy (IMN), and to explore how autophagy is related to podocyte injury, the occurrence of proteinuria and the disease progression in IMN. Methods Clinical data of 26 patients who were diagnosed as IMN (14 IMN stage 1 and 12 IMN stage 2) admitted to Zhejiang Provincial people's Hospital from January 2013 to December 2014 were retrospectively analyzed. Normal renal tissue from 15 cases of kidney neoplasms with nephrectomy was collected as control. The changes of kidney tissue pathology were detected after PAS and PASM staining by light microscope. The autophagosomes of podocyte were detected by transmission electron microscopy. Expressions of Beclin-1 and LC3 protein were detected by immunohistochemistry. Expressions of LC3 and synaptopodin were detected by immunofluorescence. The correlation of autophagosomes and clinical pathologic factors in IMN patiens was analyzed. Results There were fewer autophagosomes of podocytes and lower expression of Beclin-1 and LC3 protein in IMN group than those in control group (P=0.034, P=0.011, P=0.013, respectively). Moreover, these effects were more obvious with the development of IMN. Compared with those in control group, autophagosomes, Beclin-1 and LC3 protien were reduced in IMN stage 2 group (P=0.009, P=0.030, P=0.015); the number of autophagosomes and the expressions of LC3 and Beclin-1 were decreased in IMN stage 1 group as well, however statistically insignificant (P=0.352, P=0.087, P=0.128); Comparisons between IMN stage 2 patients and IMN stage 1 patients shown significant difference in the number of autophagosomes (P=0.030), but no significant difference in expressions of Beclin-1 and LC3 (P=0.355, P=0.181). Autophagosomes number was not correlated with serum creatinine, serum urea nitrogen, 24-hour urinary protein and eGFR (all P＞0.05). The expressions of synaptopodin and LC3 protein were lower in IMN group than those in control group. Conclusion Autophagy may contribute to podocyte injury and the production of protein urine in IMN, and may be closely related to the progression of disease.     

Role of plasma activated protein C in atherosclerosis in type 2 diabetic nephropathy

Objective To investigate the relationship between plasma activated protein C (APC) and the development of atherosclerosis (AS), and illustrate the mechanism of AS in type 2 diabetic nephropathy. Methods A total of 30 non-dialysis patients of type 2 diabetic nephrology and 26 control subjects were enrolled. APC, soluble vascular adhesion molecular-1 (sVCAM-1), soluble endothelial cell protein C receptor (sEPCR) and soluble thrombomodulin (sTM) were assayed by ELISA. Carotid intima-media thickness (IMT) was measured by ultrasonography. Results APC levels were significantly decreased in type 2 diabetic nephropathy compared with that in controls [(2 865.99±571.38) ng/L vs (3 227.70±300.44) ng/L, P＝0.005]. APC levels had negative correlation with IMT, 24 h albuminuria, sEPCR, sVCAM-1, sTM (r＝-0.720, -0.402, -0.477, -0.437, -0.505, all P＜0.05). Significant difference were observed in different proteinuria groups and IMT groups by ANOVA analysis(P＜0.05). Conclusions In type 2 diabetic nephropathy patients, plasma APC is negatively correlated with severity and IMT. Decreased plasma APC may lead to high levels of inflammatory mediators and endothelial cell injury, which may involved in the occurrence and development of atherosclerosis in diabetic nephropathy.