弹性酶降低早期糖尿病肾病尿微量白蛋白的研究

糖尿病肾病（ＤＮ）是糖尿病常见的慢性并发症，也是糖尿病致死、致残的原因之一。其主要的病理改变是肾小球基底膜增厚，系膜基质增多导致系膜扩张。临床早期表现为微量蛋白尿，如尿微量白蛋白ＡＬＢ、尿β２ＭＧ、尿Ｎ乙酰βＤ基葡萄糖苷酶（ＮＡＧ）等改变，治...     

TIMP-1、MMP-9与糖尿病肾病的关系

糖尿病肾病是糖尿病最常见、最严重的慢性微血管并发症之一,是引发终末期肾病并威胁糖尿病患者生命的主要原因,其病理改变主要为肾脏肥大,肾小管及肾小球基底膜增厚,系膜细胞增生,细胞外基质积聚致系膜区扩张,最终导致弥漫性或结节性肾小球硬化及肾小管间质纤维化。细胞外基质的堆积是导致肾小球硬化、肾间质纤维化的重要病理生理因素,细胞外基质在生理状态下处于一种快速更新代谢的动态平衡中,产生增多和（或）降解减少引起细胞外基质堆积。     

黄连素对高糖培养的大鼠肾小球系膜细胞FN及p38MAPK信号通路的影响

目的研究黄连素对高糖培养下大鼠肾小球系膜细胞细胞外基质成分纤维连接蛋白及p38MAPK信号通路的影响,进一步探讨黄连素抗糖尿病肾病的作用机制。方法实验分组:正常对照组、甘露醇组、高糖组、高糖+SB203580组、高糖+黄连素低剂量组、高糖+黄连素高剂量组共6组,观察黄连素对高糖培养下的大鼠肾小球系膜细胞纤维连接蛋白以及p38MAPK信号通路蛋白表达的影响。结果与高糖组相比,黄连素降低系膜细胞纤维连接蛋白的蛋白表达水平,抑制p38MAPK及其下游核转录因子CREB的磷酸化。结论黄连素抗糖尿病肾病的作用可能与其减少细胞外基质成分FN的积聚,抑制p38MAPK信号通路激活密切相关。     

P38 mAPK信号通路在糖尿病肾病中的作用

目的探讨p38 MAPK信号通路在糖尿病肾病中的作用。方法使用不同浓度葡萄糖及p38 mAPK特异性抑制剂SB203580干预肾小球系膜细胞,采用实时荧光定量PCR法检测各组肾小球系膜细胞p38 MAPK、MMP-9和TIMP-1 mRNA的表达。结果高糖组p38 mAPK、TIMP-1 mRNA的表达明显增加,且呈浓度依赖性;而MMP-9 mRNA表达和MMP-9/TIMP-1比值与正常组相比明显下降,同时p38 MAPK抑制剂SB203580能明显逆转以上变化。结论 p38 MAPK信号通路介导了高糖诱导的肾小球系膜细胞MMP-9和TIMP-1 mRNA表达的改变,进而参与了糖尿病肾病的进展。     

熊果酸对大鼠肾小球系膜细胞凋亡及TGF-β1/Smads信号通路的影响

目的： 观察熊果酸（ursolic acid,UA）对活化的大鼠肾小球系膜细胞（mesangial cells,MCs）凋亡与TGF-β₁/smads信号通路的影响,探讨UA抗肾纤维化的作用机制。方法： 原代培养并分离、纯化大鼠肾小球系膜细胞,转化生长因子β₁（TGF-β₁）刺激活化后,熊果酸按低剂量（2.5 μmol·L^-1）、中剂量（5 μmol·L^-1）、高剂量（10 μmol·L^-1）进行干预。Cell counting kit-8（CCK-8）及流式细胞仪分别检测系膜细胞活力及凋亡率,免疫印迹法测定系膜细胞TGF-β₁受体（TβRI、TβRII）、磷酸化蛋白P-smad信号分子表达的变化。结果： 5 μg·L^-1 TGF-β₁明显激活系膜细胞增殖,上调TGF-β₁受体表达及Smad2的磷酸化,抑制系膜细胞凋亡。低、中、高剂量熊果酸均能抑制系膜细胞增殖,促进凋亡,并下调系膜细胞TGF-β₁受体表达,抑制Smad2的磷酸化。结论： 熊果酸可通过影响TGF-β₁/Smads信号通路,诱导系膜细胞凋亡,拮抗细胞增殖,这可能是熊果酸抗肾脏纤维化的部分细胞学机制。     

糖尿病肾病中转化生长因子-β1上调与足细胞损伤

糖尿病肾病（diabeticnephropathy，DN）是糖尿病患者致死、致残的主要原因之一。尿蛋白排泄率逐年增加、肾功能持续减退至终末期是其主要临床特点。附着在肾小球基底膜（GBM）上的肾小球上皮细胞（又称足细胞，podocyte）是肾小球血液滤过屏障重要的细胞结构。转化生长因子-β1（transforminggrowthfactor—betal，TGF-β1）是一种多功能的细胞因子，在大多数细胞内起着调节生长、分化和凋亡的作用，糖尿病时高血糖和血管紧张素Ⅱ是TGF-β1的主要刺激因子。近年来研究发现，TGF-β1介导的足细胞损伤在糖尿病肾病蛋白尿发生、发展及肾小球硬化过程中起重要作用。     

大黄素治疗糖尿病肾病最新研究进展

糖尿病肾病预计在今后10年内将增加一倍,原因是发病年龄提前,存活时间延长。在美国、欧洲与日本,糖尿病肾病已成为末期肾脏病（ESRD）的主要原因（25～40%）,GFR平均每年降低12mL/min（1～24mL/min）。DN是常见的糖尿病慢性微血管并发症之一,通常指由于糖尿病引起的肾小球基底膜增厚,系膜扩张以及胞外基质增生,导致肾小球的高滤过和蛋白尿。DN是多因素引起的复杂性疾病。     

mTOR-自噬信号通路在大鼠糖尿病肾病中的作用研究

目的 研究mTOR-自噬信号通路在糖尿病肾脏损害中的作用.方法 将SD大鼠分为正常组和对照组,对照组利用佐链脲菌素构建2型糖尿病肾病模型,8周后检测两组大鼠的体重、肾重、血生化指标、尿微量白蛋白;光镜显微镜观察肾小球、肾小管形态;Western blot方法观察肾脏p-S6及LC3A/B表达.结果 成功地建立糖尿病大鼠模型,发现对照组大鼠体重减少,肾重增加,血糖、甘油三酯及尿素氮升高,而胆固醇及白蛋白有下降,尿微量白蛋白增加;光镜可见对照组大鼠肾小球肥大,系膜膜细胞增生、系膜基质增多;Western blot发现对照组大鼠肾组织的自噬活性明显降低,表现为LC3A/B降低;mTOR被激活,表现P-S6水平升高.结论 糖尿病肾病中mTOR活化,自噬能力下降,mTOR-自噬信号通路可能是糖尿病肾病的发病机制之一.     

胆汁酸膜受体TGR5对高糖培养的大鼠肾小球系膜细胞FN、TGF-β1的调控作用

目的探究G蛋白偶联受体TGR5在大鼠肾小球系膜细胞中的表达以及高糖条件下对纤维连接蛋白(fibronectin,FN)以及转化生长因子(transforming growth factor-β1,TGF-β1)蛋白水平的影响,探索其在糖尿病肾病中可能发挥的作用。方法高糖(30 mmol·L-1glucose,HG)条件下,用特异性激动剂INT-777激活TGR5,或者对TGR5进行过表达、干扰处理,通过Western blot来检测大鼠肾小球系膜细胞中TGR5的表达情况以及FN、TGF-β1蛋白水平的变化。结果在大鼠肾小球系膜细胞中存在TGR5的表达;与对照组相比,激活TGR5后,由高糖引起的FN、TGF-β1蛋白水平的上升受到抑制(P<0.01,P<0.05);另一方面,干扰TGR5后,FN、TGF-β1的蛋白水平与对照组相比异常升高,差异有统计学意义(P<0.01,P<0.05)。结论在大鼠肾小球系膜细胞中,TGR5可以抑制由高糖引起的FN、TGF-β1蛋白水平的升高,在糖尿病肾病的发生发展中可能发挥保护作用。     

骨形态发生蛋白7与糖尿病肾病

糖尿病肾病(diabetic nephropathy,DN)是糖尿病严重的微血管并发症之一,其病理特征为肾小球肥大,肾小球基底膜(glomemlar basement membrane,GBM)和肾小管基底膜(tubular basement membrane,TBM)增厚,细胞外基质(extracellular material,ECM)聚集,肾小球硬化、肾小管萎缩及间质纤维化,已经成为终末期肾病(end-stage renal disease,ESRD)死亡的主要病因[1].骨形态发生蛋白(bone morphogenetic protein,BMP)属于转化生长因子β(transforming growth factor-β,TGF-β)超家族成员之一,它是一组具有类似结构的高度保守功能蛋白,主要表现在体内诱导骨和软骨形成.目前BMP家族已发现约20多种成员,具有多种重要功能.其中,BMP-7在胚胎期对肾脏发育具有重要作用,近年来体内外研究显示BMP-7信号通路可阻断TGF-β1对肾小管上皮细胞转分化的促进作用,防治肾间质纤维化,在糖尿病肾病发生早期可能起到重要的反馈性抑制作用,延缓糖尿病肾病的发展[2].因此,深入研究BMP-7信号通路与糖尿病肾病的关系将为糖尿病肾病的治疗提供一个新思路和研究的靶点.     

GdCl3 attenuates the glomerular sclerosis of streptozotocin (STZ) induced diabetic rats via inhibiting TGF-β/Smads signal pathway

Diabetic nephropathy (DN) is the most serious end-stage renal disease which characterized by renal glomerular sclerosis including glomerular hypertrophy, glomerular basement membrane (GBM) thickening, mesangial expansion and renal fibrosis. TGF-β/Smads signal pathway plays a crucial role in the development of renal fibrosis. In this study, we found that GdCl₃ which was an agonist of Calcium-sensing receptor (CaSR) could repress the activation of TGF-β/Smads signal pathway induced by TGF-β1 or high glucose and then alleviated the accumulation of extracellular matrix (ECM) in mesangial cells and the kidney of type1 diabetic rats. Further study indicated that GdCl₃ could induce the binding of CaSR and TβR II and then both of these two receptors translocated from cell membrane to cytoplasm, in this case, TβR II on the cell membrane was decreased and then desensitized to the stimulation of its ligand TGF-β1, so that the activation of its downstream factors such as Smad2 and Smad3 were blocked, finally, ECM expression in mesangial cells were inhibited. We concluded that GdCl₃ could alleviate the accumulation of ECM in mesangial cells via antagonizing TGF-β/Smads signal pathway in diabetes mellitus.     

Suppressing renal NADPH oxidase to treat diabetic nephropathy

Renal nicotinamide adenine dinucleotide phosphate reduced form (NADPH) oxidase is an important source of oxidative stress and its expression is enhanced in the glomerulus and distal tubules of diabetic nephropathy. High glucose-induced protein kinase C signalling or renal angiotensin II signalling increases the membrane translocation of cytosolic component p47phox. NADPH oxidase-derived reactive oxygen species (ROS) in the podocytes damage the glomerular basement membrane and the slit diaphragm causing proteinuria, and mesangial and glomerular endothelial NADPH oxidase increase TGF-β and cause collagen and fibronectin accumulation. Tubular NADPH oxidase stimulated by angiotensin II or aldosterone contributes to sodium retention and to tubulointerstitial damage. Thus, inhibition of the renal renin–angiotensin II–aldosterone system with angiotensin-converting enzyme inhibitor, angiotensin II type 1 receptor blocker or selective aldosterone inhibitor indirectly suppresses NADPH oxidase reducing renal ROS, proteinuria and glomerulosclerosis. Statins are also effective in blocking the membrane translocation of Rac, especially in diabetes with hypercholesterolemia where ROS is produced by the intrinsic NADPH oxidase and by the activated macrophages. A medical herb, picrorhiza, inhibits the membrane translocation of p47phox, is a specific inhibitor of NADPH oxidase and, more so than superoxide dismutase mimetics, may be a promising strategy for the treatment of diabetic nephropathy.     

Role of AGEs in diabetic nephropathy

Diabetic nephropathy is the most common cause of end-stage renal disease in the world, and accounts for a significant increase in morbidity and mortality in patients with diabetes. Therapeutic options such as strict blood pressure and/or glycemic control are effective for preventing the development and progression of diabetic nephropathy, but the number of diabetic patients on hemodialysis is still increasing. Therefore, a novel therapeutic strategy that could halt the progression of diabetic nephropathy should be developed. Advanced glycation end products (AGEs) are heterogeneous cross-linked sugar-derived proteins which could accumulate in glomerular basement membrane, mesangial cells, endothelial cells, and podocytes in patients with diabetes and/or end-stage renal failure. AGEs are thought to be involved in the pathogenesis of diabetic nephropathy via multifactorial mechanisms such as oxidative stress generation and overproduction of various growth factors and cytokines. Further, recently, the cross-talk between AGEs and the renin-angiotensin system (RAS) has been proposed to participate in diabetic nephropathy. In addition, activation of the RAS elicits ROS generation and subsequently stimulates growth factor and cytokine production by kidney cells as well. These observations suggest that combination therapy with inhibitors of the RAS and blockers of the AGEs formation and/or their downstream pathway may be a novel therapeutic option for preventing diabetic nephropathy. In this paper, we review the role of AGEs and their receptor system in the pathogenesis of diabetic nephropathy. We further discuss here the cross-talk between AGEs and the RAS in the development and progression of diabetic nephropathy.     

糖尿病肾病相关药物研究进展

糖尿病肾病(DN)是糖尿病重要微血管并发症,其病理特征性表现为肾小球周边部位出现嗜酸性K-W结节,是糖尿病性肾病具有诊断意义的改变。但仅出现于约10%～20%的糖尿病性肾病患者。糖尿病性肾病其他病理改变特点包括肾小球基膜增厚,肾小球上皮细胞足突融合以及由于细胞外系膜基质增多所致肾小球增大。疾病晚期可出现肾小管萎缩肾内纤维化。糖尿病病程10年以上者约50%并发糖尿病肾病,每年新增终末期肾病中,糖尿病导致者所占比例逐年增高。近年关于血管紧张素转换酶抑制剂(ACEI)治疗糖尿病肾病的研究较多。其外内皮素拮抗剂等,被认为有一定的肾脏保护作用,基因治疗亦给糖尿病肾病带来新希望。现就有关内容综述如下。     

Salvianolic acid A ameliorates AGEs-induced glomerular endothelial dysfunction and protects against diabetic nephropathy

OBJECTIVE Diabetic nephropathy(DN)has been one of the most common complications of diabetes and the leading cause of end-stage renal disease.Glomerular hyperfiltrationis central in earlystage of DN and leads to the progression of renal architectonic and functional abnormalities.Salvianolic acid A(SalA)has been proved to protect diabetic complications such as hepatic fibrosis and neuropathy.The present study was designed to investigate the effects of SalA on glomerular endothelial dysfunctionand diabetic nephropathy.METHODS Primary glomerular endothelial cells were subjected to assess permeability under injury of advanced glycation end-products(AGEs).AGEs-induced changes of Rho A/ROCK pathway and cytoskeleton rearrangement were assessed bywestern blotandimmunofluorescence.The beneficial effects of SalA on diabetic nephropathy were investigated in a rat model induced by high-fat and high-glucose diet combined with low dose of streptozocin(35 mg·kg~(-1),ip).Renal function and architectonic changes were evaluated by biochemical assay and PAS staining.RESULTS SalA 3μMameliorated AGEs-induced glomerular endothelial permeability(P<0.05)and suppressed rearrangement of cytoskeleton through inhibiting AGE-RAGE-Rho A/ROCK pathway.SalA1 mg·kg~(-1)markedly reduced endothelium loss(P<0.01)and glomerular hyperfiltration(P<0.05)in diabetic kidney.Subsequently,SalA 1 mg·kg~(-1) suppressed glomerular hypertrophy and mesangial matrix expansion,eventually reduced 24 h-urinary albumin and ameliorated renal function by decreasing blood urine nitrogen(BUN),serum creatinine(Scr)and serum n-acetyl-β-d-glucosaminidase(NAG).AGEs-RAGE-Nox4-induced oxidative stress was suppressed by the treatment of SalA 1 mg·kg~(-1).CONCLUSION SalA ameliorated AGEs-induced glomerular endothelial hyperpermeability,and effectively protected against early-stage diabetic nephropathy by reducing hyperfiltration and alleviating renal structural deterioration through inhibiting AGEs and its downstream pathway.Thus,SalA might be a promising therapeutic agent for the treatment of diabetic nephropathy.     

骨形态发生蛋白-7对高糖诱导系膜细胞增生的抑制作用及其机制研究

目的 探讨骨形态发生蛋白-7(BMP-7)对高糖诱导系膜细胞增生的作用及其可能的机制.方法 体外培养大鼠肾小球系膜细胞,将细胞分为正常对照组(5.5 mmol·L-1 葡萄糖),高糖组(25 mmol·L-1),100 ng·L-1 BMP-7组,高糖+100 ng·L-1BMP-7组.应用细胞计数、MTT增殖实验及流...     

全反式维甲酸对糖尿病大鼠肾脏组织BMP-7表达及细胞外基质代谢的影响

目的探讨全反式维甲酸（atRA）对糖尿病大鼠肾脏组织骨形成蛋白-7（BMP-7）、转化生长因子-β1（TGF-β1）表达和细胞外基质（E（M）Ⅳ、Ⅲ型胶原代谢的影响。方法随机选择24只舍SD大鼠制作糖尿病模型,其中12只入糖尿病模型（DM）组,另12只入糖尿病模型＋atRA（DM＋atRA）组,此外再随机选择12只正常大鼠作为空白对照（NC）组。分别于第8、12周末各组处死6只大鼠,通过光镜、电镜观察各组大鼠肾脏病理结果;通过免疫组织化学方法观察BMP-7,TGF-β1,Ⅳ、Ⅲ型胶原表达,并进行半定量分析;通过实时荧光定量PCR方法对TGF-β1 mRNA、BMP-7mRNA,进行相对定量分析。结果DM＋atRA组大鼠肾组织BMP-7表达较同时期DM组增加（P〈0.01）,主要表达于大鼠肾小管上皮细胞,而TGF-β1表达较同时期DM组减少（P〈0．01）：且DM＋atRA组大鼠肾组织ECM重要成分Ⅳ、Ⅲ型胶原表达与DM组相比较显著减少（P〈0．01）。同时观察到BMP-7蛋白与Ⅳ型胶原蛋白表达呈负相关（r=-0．75,P〈0．01）,与Ⅲ型胶原蛋白表达呈负相关（r=-0．71,P〈0．01）,与TGF-β1蛋白表达呈负相关（r=-0．86,P〈O．01）。荧光定量PCR结果表明,DM＋atRA组BMP-7mRNA水平升高,TGF-β1 mRNA水平下降,且两者星显著负相关（r=-0．83,P〈0．01）。结论atRA能减轻糖尿病大鼠肾脏病理损伤,延缓肾脏纤维化,这一防治作用可能与其增加BMP-7表达,下调TGF-β1表达从而减少肾组织ECM沉积有关。     

姜黄素对糖尿病大鼠肾脏TGF-β1影响及保护性研究

目的观察糖尿病大鼠肾脏TGF—β1表达及姜黄素胶囊的治疗作用。方法采用STZ注射法制作糖尿病（DM）大鼠模型,随机分为正常组,DM组,姜黄素组。灌胃给药8周后,采用免疫组化等方法测定24h尿白蛋白排泄率（UAER）、肾小球滤过率（GFR）、肾皮质TGF-β1、FN、Col—Ⅳ的表达。结果（1）功能学变化：姜黄素组大鼠的24hUAER和GFR显著低于DM、对照组,差异有统计学意义（P〈0．05）。（2）免疫组化：姜黄素组大鼠肾皮质TGF—β1、FN、Col—Ⅳ的表达显著低于DM组,差异有统计学意义（P〈0．05）。结论姜黄素胶囊有降低早期糖尿病肾病大鼠尿白蛋白排泄率及减少过高的GFR、抑制‘肾组织增生、抑制。肾皮质TGF-β1、FN、Col—Ⅳ表达等保护作用。