Calcineurin (CaN) plays an important role in glomerular hypertrophy and extracellular matrix accumulation in early diabetic nephropathy. Cyclosporine (CSA), a CaN inhibitor, has been shown to reduce renal injury in streptozotocin-induced diabetic rats. We examined whether FK506, which immunosuppressive action was 10-100 times of CSA, inhibits progression of diabetic nephropathy in experimental diabetic rats. Diabetes was induced with streptozotocin in rats, and FK506 (0.5 or 1.0mg/kg) was orally administered once a day for 4 weeks. Increased relative kidney weight was significantly reduced by FK506 treatment with 1.0mg/kg (p<0.05), and elevated 24 hour urinary albumin excretion rate was markedly attenuated by FK506 treatment with 0.5 and 1.0mg/kg (p<0.05, 0.01). Elevated glomerular volume was significantly attenuated by FK506 treatment with 0.5 and 1.0mg/kg (p<0.05), and increased indices for tubulointerstitial injury were only ameliorated by FK506 treatment with 1.0mg/kg (p<0.01). Western blot analysis noted that the expression of CaN protein was increased 2.4 fold in the kidney from diabetic rats, and FK506 treatment with 0.5 and 1.0mg/kg could reduce increased expression of CaN protein by 38.0% and 73.2%. The expression of 1α (IV) collagen, p65, p-p65, OPN, α-SMA and TGF-β1 protein in kidney was significantly increased in diabetic rats and reduced by FK506 treatment (p<0.05, 0.01). Our results show that FK506 could ameliorate renal injury in early experimental diabetic rats, which mechanism may be at least partly correlated with suppression on increased CaN in renal tissue in diabetic rats.

【Objective】To investigate the effect of TGP on activation of JAK/STAT signal transduction in the kidney from diabetic rat and explore the possible renoprotection mechanisms. 【Methods】Diabetes was induced with streptozotocin in rats, and TGP was orally administered once a day for 8 weeks to rats. The expression of the p- JAK2, p-STAT3 and 1α (IV) collagen protein were determined by western blot analysis and expression of TGF-β1 was measured by immunohistochemistry in the kidney. 【Results】Elevated 24 hours urinary albumin excretion rate was markedly attenuated by TGP treatment. Western blot analysis and immunohistochemistry noted that the expression of p-JAK2, p-STAT3, 1α (IV) collagen protein and TGF-β1 protein in the kidney were significantly increased in diabetic rats, while they were significantly inhibited by TGP treatment. 【Conclusion】Our data suggest that TGP treatment ameliorates early renal injury via the inhibition of activation of JAK/STAT signal transduction and the relevant TGF-β1-collagen IV expression in the kidney from diabetic rat.

Total glucosides of paeony (TGP), extracted from the root of Paeonia lactiflora pall, has been shown to have ant-inflammatory and antioxidative actions. The aims of this study were to elucidate the renoprotective effect of TGP and its mechanism in experimental diabetes. Streptozotocin-induced diabetic rats were treated with TGP for 8 weeks. Treatment with TGP at 50, 100, and 200 mg/kg significantly lowered 24-h urinary albumin excretion rate in diabetic rats. TGP treatment in all doses markedly attenuated glomerular volume, and treatment with TGP at 100 and 200 mg/kg markedly reduced indices for tubulointerstitial injury in diabetic rats. Western blot analysis showed that the expressions of 1 alpha (IV) collagen, intercellular adhesion molecule (ICAM)-1, interleukin (IL)-1, tumor necrosis factor (TNF)-alpha, NF-kappaB p65, and 3-nitrotyrosine (3-NT) protein were increased in the kidneys of diabetic rats; the increases in these proteins were all dose-dependently and significantly inhibited by TGP treatment. The expression of nephrin protein was significantly reduced in the kidneys from diabetic rats and markedly increased by TGP treatment. The expression of transforming growth factor (TGF)-beta1 protein in the kidney was also significantly increased in diabetic rats, which was significantly inhibited by treatment with TGP at all doses. Our data suggest that TGP treatment ameliorates early renal injury via the inhibition of expression of ICAM-1, IL-1, TNF-alpha, and 3-NT in the kidneys of diabetic rats.

目的探讨白芍总苷（TGP）对糖尿病肾病的治疗作用及其可能的作用机制。方法采用链佐星（STZ）诱导大鼠糖尿病模型。大鼠随机分为对照组、糖尿病模型组、TGP组（50,100和200mg.kg-1,ig,每天1次,共8周）。8周后检测肾组织总抗氧化能力（T-AOC）、超氧化物歧化酶（SOD）、过氧化氢酶（CAT）和谷胱甘肽过氧化物酶（GSH-PX）活性,应用Western蛋白印迹法检测肾组织硝基酪氨酸（NT）蛋白的表达,免疫组化方法检测肾组织转化生长因子β1（TGFβ1）蛋白的表达。结果与对照组相比,模型组肾组织T-AOC,SOD和CAT活性明显降低;TGP200mg.kg-1给药组T-AOC,SOD和CAT活性明显高于模型组。模型组肾组织NT蛋白表达较对照组增加3.4倍,给予TGP50,100和200mg.kg-18周可分别使肾组织NT蛋白表达下降41.2%,43.8%和57.5%。模型组肾组织TGFβ1蛋白表达明显高于对照组,TGP50,100和200mg.kg-1组TGFβ1蛋白表达明显低于模型组。结论糖尿病大鼠肾脏存在氧化应激反应,TGP抗糖尿病肾病的作用可能与其抗氧化活性有关。

Several works in the setting of early experimental diabetic nephropathy using anti-inflammatory drugs, such as mycophenolate mofetil (MMF), have shown that prevention of the development or amelioration of renal injury including proteinuria. The exact mechanisms by which anti-inflammatory drugs lower the albuminuria have no still to clarify well. In this study, diabetes was induced by injection of streptozotocin after uninephrectomy. Rats were randomly divided into three groups: control group, diabetic group and diabetic group treated with MMF. Elevated 24h urinary albumin excretion rate was markedly attenuated by MMF treatment. In diabetic rats receiving no treatment, there were increase in ED-1+ cells in the glomeruli, which were effectively suppressed by MMF treatment. The expression of nephrin and podocin protein was reduced in the glomeruli from diabetic rats, and MMF treatment significantly increased the expression of nephrin and podocin. The expression of IL-1, TNF-α and 3-NT protein in the glomeruli were significantly increased in diabetic rats, which were all significantly inhibited by MMF treatment. Our results show that MMF could decrease urinary albumin excretion, which mechanism may be at least partly correlated with upregulated expression of nephrin and podocin in the glomeruli of diabetic rat.

Aim To investigate the effect of mycophenolate mofetil(MMF)on the expression of Nephrin and Podocin and its mechanism in the kidney of diabetic rats.Methods Diabetes was induced with streptozotocin after uninephrectomy,and MMF(10 mg·kg-1·d-1 was orally administered once a day for 8 wk.Blood glucose and 24 hours urinary albumin excretion rate(AER)were measured.The expression of ED-1,Nephrin,Podocin,interleukin-1(IL-1)and tumor necrosis factor-α(TNF-α)protein in the kidney were determined by immunohistochemistry or Western blot analysis.Results Elevated AER was markedly attenuated by MMF treatment(P0.05).ED-1-positive cells were significantly increased in glomeruli of diabetic rats,which was effectively suppressed by MMF treatment(P0.05).Western blot analysis showed that the expression of Nephrin and Podocin protein was reduced in the kidney of diabetic rats,and MMF treatment significantly increased the expression of Nephrin and Podocin protein(P0.01).The expression of IL-1 and TNF-α protein in the kidney was significantly increased in diabetic rats,which was significantly inhibited by MMF treatment(P0.01).Conclusions MMF could decrease AER in diabetic rats,whose mechanism may be at least partly correlated with upregulating the expression of Nephrin and Podocin in the kidney.

炎性反应在糖尿病肾病(DN)的发生和进展中起着十分重要的作用[1,2].白芍总苷(TGP)是从我国传统中药白芍根中提取的有效成分,其药理作用主要有抗炎、抗氧化与免疫调节活性.我们既往研究表明TGP对糖尿病大鼠肾脏有明显保护作用[3].本研究观察了TGP对糖尿病模型肾内白细胞介素1(IL-1)与肿瘤坏死因子α(TNF-α)表达的影响,旨在进一步探讨TGP对糖尿病肾脏的保护作用机制.

目的 探讨FK506对糖尿病大鼠早期肾脏肥大的抑制作用及机制.方法 应用链脲菌素(65 mg/kg)腹腔注射建立大鼠糖尿病模型.每日分别给予FK506 0.5、1.0 mg/kg灌胃,共4周:观察大鼠肾质量/体质量、尿白蛋白排泄率(AER)、Ccr及肾组织病理形态学变化.应用Western印迹和免疫组化方法检测肾组织钙神经蛋白(calcineurin,CaN)、1αⅣ型胶原、α-平滑肌肌动蛋白(α-SMA)及转化生长冈子β1(TGF-β1)蛋白表达.结果 FK506 1.0组大鼠相对肾质量明显低于模型组(P＜0.05).FK506 0.5与1.0组大鼠AER水平与肾小球平均体积明显低于模型组(P＜0.05、0.01).FK506 1.0组肾小管间质-损伤指数也明显低于模型组＜P＜0.01).Western印迹显示,模型组肾组织CaN蛋白表达较对照组增加2.4倍;FK506 0.5与1.0组肾组织CaN蛋白表达比模型组下降38.0%与73.2%.模型组肾组织1αⅣ型胶原、Ⅳ-SMA及TGF-β1蛋白表达明显高于对照组;FK506组肾组织上述蛋白表达明显低于模型组(P＜0.05、0.01).结论 FK506对糖尿病大鼠早期肾脏肥大有明显抑制作用,其机制与下调肾组织增高的CaN表达有关.

目的 探讨麦考酚吗乙酯(MMF)对糖尿病大鼠肾组织白细胞介素-1(IL-1)与肿瘤坏死因子-α(TNF-α)表达的影响.方法 建立链脲佐菌素诱导的单侧肾切除大鼠糖尿病模型,随机分成对照组(C组)、糖尿病组(DM组)与MMF给药组[DM+MMF组,10 mg/(kg*d)灌胃给药].8周末观察血糖、24 h尿白蛋白排泄率(AER)及肾小球病理形态学变化,Western印迹方法检测肾组织IL-1与TNF-α蛋白表达.结果 DM组大鼠尿AER明显高于C组(P<0.01),DM+MMF组大鼠尿AER明显低于DM组(P<0.05) .DM组肾小球面积(AG)、系膜区面积(AM)、肾小球容积(VG)明显高于C组(P<0.05, P<0.01),MMF可明显抑制AG、VG、AM的增加(P<0.05).Western印迹显示DM组肾组织IL-1,TNF-α蛋白表达明显高于C组,MMF给药8周明显降低它们的表达(P<0.01).结论 MMF对糖尿病大鼠肾脏有保护作用,其机制可能部分与其抑制肾组织过度表达的IL-1与TNF-α蛋白有关.

Aim To investigate the effect of total glucosides of paeony (TGP) on the expression of intercellular adhesion molecule-1 (ICMA-1) and transforming growth factor 1 (TGF 1) protein in the kidney in experimental diabetes. Methods Diabetes was induced with streptozotocin in Sprague-Dawley rats. TGP (50, 100, 200 mg·kg-1·d-1) was orally administered once a day for 8 wk to rats. Results Elevated urinary albumin excretion rate (AER) was markedly attenuated by TGP treatment with 50, 100 and 200 mg·kg-1. Increased malondialdehyde (MDA) level was only significantly reduced by TGP treatment with 200 mg·kg-1. Elevated glomerular volume was significantly attenuated by TGP treatment with 50, 100 and 200 mg·kg-1, and increased indices for tubulointerstitial injury were only ameliorated by TGP treatment with 100 and 200 mg·kg-1. Western blot analysis noted that the expression of 1 type Ⅳ collagen protein was increased 2.7 -fold in the kidney in diabetic rats, TGP treatment with 50, 100 and 200 mg·kg-1 could reduced increased expression of 1 type Ⅳ collagen protein by 47.9 %, 60.4 % and 72.9 %. The expression of ICAM-1 and TGF 1 protein in the kidney were significantly increased in diabetic rats, which were all significantly inhibited by TGP treatment. Conclusion Our data suggest that TGP treatment ameliorates early renal injury via the inhibition of oxidative stress and overexpression of ICAM-1 and TGF-1 in renal tissue in diabetic rats.