Shenkang injection (SKI) is a classic prescription composed of Radix Astragali, rhubarb, Astragalus, Safflower, and Salvia. This treatment was approved by the State Food and Drug Administration of China in 1999 for tr...Shenkang injection (SKI) is a classic prescription composed of Radix Astragali, rhubarb, Astragalus, Safflower, and Salvia. This treatment was approved by the State Food and Drug Administration of China in 1999 for treatment of chronic kidney diseases based on good efficacy and safety. This study aimed to investigate the protective effect of SKI against high glucose (HG)-induced renal tubular cell senescence and its underlying mechanism. Primary renal proximal tubule epithelial cells were cultured in (1) control medium (control group), medium containing 5 mmol/L glucose;(2) mannitol medium (mannitol group), medium containing 5 mmol/L glucose, and 25 mmol/L mannitol;(3) HG medium (HG group) containing 30 mmol/L glucose;(4) SKI treatment at high (200 mg/L), medium (100 mg/L), or low (50 mg/L) concentration in HG medium (HG+ SKI group);or (5) 200 mg/L SKI treatment in control medium (control+ SKI group) for 72 h. HG-induced senescent cells showed the emergence of senescence associated heterochromatin foci, up-regulation of P16INK4 and cyclin D1, increased senescence-associated β-galactosidase activity, and elevated expression of membrane decoy receptor 2. SKI treatment potently prevented these changes in a dose-independent manner. SKI treatment prevented HG-induced up-regulation of pro-senescence molecule mammalian target of rapamycin and p66Shc and down-regulation of anti-senescence molecules klotho, sirt1, and peroxisome proliferator-activated receptor-g in renal tubular epithelial cells. SKI may be a novel strategy for protecting against HG-induced renal tubular cell senescence in treatment of diabetic nephropathy.展开更多
Sustained proteinuria is an independent risk factor leading to kidney fibrosis and end-stage renal fail-ure.Over-reabsorption of filtered proteins,notably albu-min,has been proved to trigger interstitial inflammation ...Sustained proteinuria is an independent risk factor leading to kidney fibrosis and end-stage renal fail-ure.Over-reabsorption of filtered proteins,notably albu-min,has been proved to trigger interstitial inflammation and fibrosis in proteinuric renal disease.Cubilin,an endo-cytic receptor expressed on the renal tubular brush bor-der,is responsible for albumin reabsorption in physiologic condition.However,little is known about whether it is required for activation of tubular cells induced by albu-min overload.In this work,we investigated the change of cubilin expression and its potential role in albumin-induced up-regulation of chemokines synthesis in vivo and in vitro.Twenty-six patients with nephrotic syndrome were enrolled in this study.Proximal tubule uptake of albumin,expression of apical membrane cubilin and infiltrating cells in kidney interstitium were determined by immunocytochemistry.In vitro,the transcription of cubilin in HK2 cells after exposure to albumin was ana-lyzed by real-time PCR.Endocytosis of albumin in HK2 cells was examined by fluorescent microscope.The influ-ence of inhibition of cubilin on albumin-induced expres-sions of monocyte chemoattractant protein 1(MCP-1)and regulated upon activation normal T-cell expressed and secreted(RANTES)was investigated by Western blot.The intensity of luminal cubilin and tubular accu-mulation of albumin were significantly increased in nephrotic kidneys.The expression of MCP-1 and RANTES was up-regulated,and there were spatial rela-tionships in localization between these chemokines and cubilin as well as intracellular albumin in kidney tissues.Infiltration of CD-3 and ED-1-positive cells was predom-inant in tubulointerstitial areas displaying signs of increases of cubilin expression and albumin accumula-tion.In vitro,the transcription of cubilin mRNA in HK2 cells was enhanced after 24 h exposure to albumin in a dose-dependent manner.Inhibition of endocytosis of albumin by antisense cubilin nucleotide markedly reduced expression of MCP-1 and RANTES.Cubilin was required for handling a greater amount of protein in nephrotic status and albumin-induced production of MCP-1 and RANTES by renal tubular cells,which further initiated tubulointerstitial inflammation in proteinuric disease.展开更多
文摘Shenkang injection (SKI) is a classic prescription composed of Radix Astragali, rhubarb, Astragalus, Safflower, and Salvia. This treatment was approved by the State Food and Drug Administration of China in 1999 for treatment of chronic kidney diseases based on good efficacy and safety. This study aimed to investigate the protective effect of SKI against high glucose (HG)-induced renal tubular cell senescence and its underlying mechanism. Primary renal proximal tubule epithelial cells were cultured in (1) control medium (control group), medium containing 5 mmol/L glucose;(2) mannitol medium (mannitol group), medium containing 5 mmol/L glucose, and 25 mmol/L mannitol;(3) HG medium (HG group) containing 30 mmol/L glucose;(4) SKI treatment at high (200 mg/L), medium (100 mg/L), or low (50 mg/L) concentration in HG medium (HG+ SKI group);or (5) 200 mg/L SKI treatment in control medium (control+ SKI group) for 72 h. HG-induced senescent cells showed the emergence of senescence associated heterochromatin foci, up-regulation of P16INK4 and cyclin D1, increased senescence-associated β-galactosidase activity, and elevated expression of membrane decoy receptor 2. SKI treatment potently prevented these changes in a dose-independent manner. SKI treatment prevented HG-induced up-regulation of pro-senescence molecule mammalian target of rapamycin and p66Shc and down-regulation of anti-senescence molecules klotho, sirt1, and peroxisome proliferator-activated receptor-g in renal tubular epithelial cells. SKI may be a novel strategy for protecting against HG-induced renal tubular cell senescence in treatment of diabetic nephropathy.
文摘Sustained proteinuria is an independent risk factor leading to kidney fibrosis and end-stage renal fail-ure.Over-reabsorption of filtered proteins,notably albu-min,has been proved to trigger interstitial inflammation and fibrosis in proteinuric renal disease.Cubilin,an endo-cytic receptor expressed on the renal tubular brush bor-der,is responsible for albumin reabsorption in physiologic condition.However,little is known about whether it is required for activation of tubular cells induced by albu-min overload.In this work,we investigated the change of cubilin expression and its potential role in albumin-induced up-regulation of chemokines synthesis in vivo and in vitro.Twenty-six patients with nephrotic syndrome were enrolled in this study.Proximal tubule uptake of albumin,expression of apical membrane cubilin and infiltrating cells in kidney interstitium were determined by immunocytochemistry.In vitro,the transcription of cubilin in HK2 cells after exposure to albumin was ana-lyzed by real-time PCR.Endocytosis of albumin in HK2 cells was examined by fluorescent microscope.The influ-ence of inhibition of cubilin on albumin-induced expres-sions of monocyte chemoattractant protein 1(MCP-1)and regulated upon activation normal T-cell expressed and secreted(RANTES)was investigated by Western blot.The intensity of luminal cubilin and tubular accu-mulation of albumin were significantly increased in nephrotic kidneys.The expression of MCP-1 and RANTES was up-regulated,and there were spatial rela-tionships in localization between these chemokines and cubilin as well as intracellular albumin in kidney tissues.Infiltration of CD-3 and ED-1-positive cells was predom-inant in tubulointerstitial areas displaying signs of increases of cubilin expression and albumin accumula-tion.In vitro,the transcription of cubilin mRNA in HK2 cells was enhanced after 24 h exposure to albumin in a dose-dependent manner.Inhibition of endocytosis of albumin by antisense cubilin nucleotide markedly reduced expression of MCP-1 and RANTES.Cubilin was required for handling a greater amount of protein in nephrotic status and albumin-induced production of MCP-1 and RANTES by renal tubular cells,which further initiated tubulointerstitial inflammation in proteinuric disease.
