The effective bioactivity compositions of uremic clearance granul (UCG) include isoflavonoids, emodin, astragaloside, paeoniflorin, salvianolic acid A, and so on. The effects of UCG treating chronic renal failure (CRF) in clinical pharmacodynamics mainly refer to improve renal function and the complications of CRF. The mechanisms involved in vivo basically include depressing transforming growth factor (TGF)-beta1 over-expression, lessening podocyte injury,inhibiting tubular epithelial myofibroblast transdifferentiation, ameliorating microinflammation status, retarding oxidative stress, and alleviating insulin resistance.
Animals ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Kidney ; drug effects ; metabolism ; Kidney Failure, Chronic ; drug therapy ; genetics ; metabolism ; Oxidative Stress ; drug effects ; Transforming Growth Factor beta1 ; genetics ; metabolism

OBJECTIVETo explore the effect of resveratrol on transforming growth factor-beta1 (TGF-beta1) induced transdifferentiation of podocytes.

METHODSMouse podocytes in vitro cultured under differentiating conditions for 10 days were divided into the normal group, the model group, the high dose resveratrol group, and the low dose resveratrol group. The podocytes in the high and low dose resveratrol groups were intervened with 5 micromol/L and 2 micromol/L resveratrol respectively for 30 min. Those in the model group and the two resveratrol treated groups were continually incubated with 5 ng/mL TGF-beta1 for 72 h. Those in the normal group were routinely cultured. The protein expression of podocyte phenotypic protein molecules such as E-cadherin, P-cadherin, zonula occludens-1 (ZO-1), NEPH1, and alpha-smooth muscle-actin (alpha-SMA) were detected by immunocytochemistry, flow cytometry (FCM), and Western blot. A simple albumin influx assay was used to evaluate the filtration barrier function of podocyte monolayer.

RESULTSCompared with the normal control group, E-cadherin (+) percentage rate, the protein expression of P-cadherin, ZO-1, and NEPH1 significantly decreased in the model group (P < 0.05), but the expression of alpha-SMA and albumin permeability across podocyte monolayers increased significantly (P < 0.05). Compared with the model group, E-cadherin (+) percentage rate significantly increased (P < 0.05) and albumin permeability across podocyte monolayers decreased significantly (P < 0.05) in the high and low dose resveratrol groups. In the low dose resveratrol group, the expression of P-cadherin and NEPH1 significantly increased (P < 0.05). In the high dose resveratrol group, the expression of P-cadherin, ZO-1, and NEPH1 increased significantly, and the expression of alpha-SMA decreased significantly (P < 0.05). The correlations between resveratrol concentrations and the expression of E-cadherin (+), P-cadherin, and NEPH1 were significantly positive (r(E-cadherin (+)) = 0.772, r(P-cadherin) = 0.756, r(NEPH1) = 0.809, P < 0.05).

CONCLUSIONThe role of resveratrol in inhibiting TGF-beta1 induced phenotype abnormality might be an important mechanism for preserving the integrality of glomerular filtration barrier and decreasing proteinuria.

Animals ; Cell Transdifferentiation ; drug effects ; Cells, Cultured ; Kidney Tubules ; cytology ; drug effects ; Mice ; Podocytes ; cytology ; drug effects ; Stilbenes ; pharmacology ; Transforming Growth Factor beta1 ; metabolism

In this review,firstly,it has been discussed the mechanisms of Chinese herbal medicine ameliorating glomerulosclerosis and renal interstitial fibrosis during the progression of chronic renal failure (CRF) by improving glomerular hemodynamics turbulence, podocyte injury, transforming growth factor (TGF)-beta over-expression, hyperlipidemia, macrophage infiltration, tubular epithelial myofibroblast transdifferentiation, and nephrotoxicity of proteinuria. Secondly,it has been reported the clinical effects of Chinese herbal medicine improving renal function and some clinical complications in the patients with progressive CRF through various treatments including oral administration or coloclysis of Chinese herbal medicine, oral administration combined with coloclysis of Chinese herbal medicine, and colonic dialysis combined with coloclysis of Chinese herbal medicine. Finally,it has been reviewed the beneficial influences of Chinese herbal medicine on metabolic dysequilibrium of calcium and phosphonium, microinflammatory state, and uremic toxins in patients with uremia.
Animals ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Kidney ; drug effects ; metabolism ; pathology ; Kidney Failure, Chronic ; drug therapy ; Transforming Growth Factor beta ; metabolism

Cell Differentiation ; drug effects ; Cell Line ; Connective Tissue Growth Factor ; metabolism ; Epithelial Cells ; cytology ; drug effects ; metabolism ; Epithelial-Mesenchymal Transition ; drug effects ; Humans ; Kidney Tubules, Proximal ; cytology ; Lead ; toxicity ; Transforming Growth Factor beta ; metabolism

OBJECTIVETo investigate the effects of microRNA-145 (miR-145) on epithelial-mesenchymal transition (EMT) of TGF-β1-induced human renal proximal tubular epithelial (HK-2) cells.

METHODSThe gene sequence of miR-145 was synthesized and cloned into pCMV-myc to construct recombinant plasmid pCMV-miR-145. HK-2 cells were divided into four groups: control (untreated), TGF-β1 (treated with TGF-β1), blank+TGF-β1 (treated with TGF-β1 after HK-2 cells transfected with blank plasmid) and miR-145+TGF-β1 (treated with TGF-β1 after HK-2 cells transfected with pCMV-miR-145 recombinant plasmid). Expression of miR-145 was detected by real-time PCR (RT-PCR). TGF-β1, Smad3, Smad2/3, p-Smad2/3, α-SMA, FN and type I collagen (Col I) protein levels were detected by Western blot. Concentrations of fibronectin (FN) and Col I in cell culture supernatants were measured using ELISA.

RESULTSpCMV-miR-145 recombinant plasmid was successfully transfected into HK-2 cells. Compared with the control group, the miR-145+TGF-β1 group showed a significant up-regulation in the expression level of miR-145 (P<0.01). However, the TGF-β1 and blank+TGF-β1 groups showed a significant down-regulation in the expression level of miR-145 compared with that in the control and miR-145+TGF-β1 groups (P<0.01). Compared with the TGF-β1 and blank+TGF-β1 groups, the miR-145+TGF-β1 group showed significantly reduced levels of the signal proteins TGF-β1, Smad3, Smad2/3 and p-Smad2/3 (P<0.05), as well as significantly reduced levels of the biomarkers α-SMA, FN and Col I (P<0.05). Meanwhile, concentrations of FN and Col I in cell culture supernatants also decreased (P<0.05).

CONCLUSIONSmiR-145 modulates the EMT of HK-2 cells treated with TGF-β1, possibly by inhibition of the activation of TGF-β-dependent Smad signaling pathway.

Cells, Cultured ; Epithelial Cells ; drug effects ; pathology ; Epithelial-Mesenchymal Transition ; Humans ; Kidney Tubules, Proximal ; drug effects ; pathology ; MicroRNAs ; physiology ; Transforming Growth Factor beta1 ; pharmacology

OBJECTIVESTo explore the effects and significance of Huanshuai Recipe Oral Liquid (, HSR), a formula with supplementing qi, nourishing blood and activating blood on restructuring glomerular microvasculature and expression of vascular endothelial growth factor (VEGF) in subtotal nephrectomized (SNX) rats.

METHODSA total of 76 male Wistar rats were randomly divided into four groups: 16 in the sham-operated group and fed with tap water 10 mL/kg per day; 20 in the model group were operated with 5/6 SNX and fed with tap water 10 mL/ kg per day; 20 SNX rats in the HSR group were treated with HSR 10 mL/kg per day; 20 SNX rats in the losartan group were treated with losartan 40 mg/kg per day. Serum creatinine (SCr) and urinary protein excretion (Upro) were examined at the 2nd, 4th, 8th, and 12th weeks of the treatment, and the remnant kidneys were harvested. Changes in histological microstructure were evaluated using light microscopy, and the expression of VEGF was detected by using ELISA.

RESULTSUpro, microvasculature injury and glomerulosclerosis were found to be alleviated in HSR and Losartan groups, respectively. The change of VEGF expression showed positive correlation with glomerular capillary area and peritubular capillary number (r=0.448, r=0.422, P<0.01), but negative correlation with that of SCr and Upro (r=-0.592, r=-0.481, P<0.01).

CONCLUSIONSHSR could regulate the VEGF expression, reduce the loss of microvasculature, which demonstrated similar renal protective effects to losartan in SNX rats. Examination of Chinese herbal medicine influence on VEGF signaling and restructuring renal microvasculature may elucidate the molecular mechanism of renal protection to a certain degree.

Administration, Oral ; Animals ; Capillaries ; drug effects ; metabolism ; pathology ; Collagen Type IV ; metabolism ; Creatinine ; blood ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; Extracellular Matrix ; drug effects ; metabolism ; Fibronectins ; metabolism ; Immunohistochemistry ; Kidney Cortex ; drug effects ; metabolism ; pathology ; Kidney Function Tests ; Kidney Glomerulus ; blood supply ; drug effects ; pathology ; physiopathology ; Microvessels ; drug effects ; Nephrectomy ; Proteinuria ; blood ; drug therapy ; physiopathology ; Rats ; Rats, Wistar ; Time Factors ; Vascular Endothelial Growth Factor A ; metabolism

OBJECTIVETo study the effects of core proteoglycan on the transdifferentiation of human renal tubular epithelial cell induced by transforming growth factor beta1 (TGF-beta1) in vitro.

METHODThe cultured HK-2 cells were divided into six groups: A. negative control group; B. 10 ng/ml TGF-beta1 group; C. 10 ng/ml core proteoglycan treated group; D. 100 ng/ml core proteoglycan treated group; E. 10 ng/ml TGF-beta1 + 10 ng/ml core proteoglycan group; F. 10 ng/ml TGF-beta1 + 100 ng/ml core proteoglycan group. The changes in configuration of HK-2 cells were inspected 48 hours after adding the stimulating factor. At the same time, changes in mRNA of keratin, alpha-smooth muscle actin, vimentin were analyzed.

RESULTSCompared with group A, group B showed great changes in the morphology of cells, most cells converted into spindle shape, like fibroblast; groups E and F, especially group F showed significantly reduced spindle shape cells. Compared with group A, groups C and D had no significant changes in morphology of cells Compared with 10 ng/ml TGF-beta1 group and negative control, the mRNA expression of alpha-smooth muscle actin and vimentin had significant increase, but that of keratin reduced (P < 0.05). However, after combined treatment with TGF-beta1 and core proteoglycan, alpha-smooth muscle actin and vimentin expression were reduced significantly, while expression of keratin was up-regulated. Single core proteoglycan treated group and negative control group had no dramatic differences (P > 0.05).

CONCLUSIONTGF-beta1 can induce the transdifferentiation of human renal tubular epithelial cell and core proteoglycan has some inhibitory effect on transdifferentiation of human renal tubular epithelial cell induced by TGF-beta1 in vitro.

Actins ; physiology ; Bone Morphogenetic Protein 7 ; metabolism ; Cadherins ; metabolism ; Cell Differentiation ; drug effects ; physiology ; Cell Line ; Cell Shape ; Cell Transdifferentiation ; Cells, Cultured ; Connective Tissue Growth Factor ; pharmacology ; Epithelial Cells ; cytology ; drug effects ; physiology ; Fibroblasts ; drug effects ; pathology ; Humans ; Kidney ; pathology ; Kidney Tubules ; pathology ; Kidney Tubules, Proximal ; pathology ; Proteoglycans ; chemistry ; pharmacology ; Transforming Growth Factor beta ; genetics ; pharmacology ; Transforming Growth Factor beta1 ; chemistry ; pharmacology ; Vimentin ; metabolism

OBJECTIVETo investigate the effects of ginsenoside R(g1) on the transdifferentiation of rat renal tubular epethelial cells induced by transforming growth factor-beta1, (TGF-beta1).

METHODCultured normal rat renal tubular epethelial cells (NRK-52E) were divided into control group, TGF-beta1-induced group and treated with ginsenoside R(g1) at different concentration (10, 20, 40 mg x L(-1)) group. The morphology of tubular epithelial-myofibroblast transdifferentiation induced by TGF-beta1 was observed through light microscope. alpha-SMA and E-cadherin protein expression were assessed by immunohistochemistry and western blot analyses. alpha-SMA, collagen I and and fibronectin gene expression were assessed by real-time quantitative chain reaction. Enzyme-linked immunosorbent assay was used to quantitatively detect collagen I and fibronectin in the supernatant.

RESULT10 mg x L(-1) TGF-beta1 could induce the transdifferentiation of tubular epithelial myofibroblast, showing fibroblast-like in morphology, with significantly enhanced expression of alpha-SMA, depressed expression of E-cadherin and increased secretion of fibronectin and collagen I (P < 0.05). Compared to TGF-beta1-induced group, ginsenoside R(g1) partly abrogated the alpha-SMA expression and E-cadherin depression triggered by TGF-beta1 in tubular epithelial cells in a dose-dependent manner (P < 0.05). Meanhile, ginsenoside R(g1) blocked morphologic transformation of tubular epithelial cells and decreased levels of collagen I and fibronectin (P < 0.05).

CONCLUSIONGinsenoside R(g1) could inhibit TGF-beta1 induced the tubular epithelial-myofibroblast transdifferentiation and decreased levels of collagen I and fibronectin in NRK52E.

Animals ; Cadherins ; genetics ; metabolism ; Cell Line ; Cell Transdifferentiation ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Epithelial Cells ; cytology ; drug effects ; Gene Expression ; drug effects ; Ginsenosides ; pharmacology ; Kidney Tubules ; cytology ; drug effects ; Panax ; chemistry ; Rats ; Transforming Growth Factor beta1 ; pharmacology

OBJECTIVEThe purpose of this study was to investigate the effects of exogenous epidermal growth factor (EGF) on the growth inhibition of renal proximal tubular epithelial cell induced by AA-I.

METHODCultured human renal proximal tubular epithelial cell line HK-2 was used as the subject. The changes of the survived HK-2 cells were observed and compared among control, AA-I stimulation, pre-EGF, together-EGF and post-EGF groups. In the study, cellular morphologic assessments were performed with a phase-contrast inverted microscope. Cell counting after stained with 0.04% trypan blue was adopted to analyze cell proliferation. Cell cycle was assessed by flow cytometry.

RESULTNumber of the survived cells stimulated by AA-I for 12, 24, 48 hours decreased gradually in a dose and time dependent manner. At 24, 48 hours, the survived cells showed a significant disturbance in the cell cycle procedure, which was characterized as decreased percentage of cells in G0/G1 phase, significant increased percentage of cells in G2/M phases. Exogenous EGF (20 microg L(-1)) could significantly promote the proliferation of HK-2 cells, which shown a increased cell number, accompanied down-regulated cells in G0/G1 phase, increased cells in S and G2/M phase. Compared with AA-I groups, it failed to improve the inhibitory effect on cell proliferation and abnormal cell cycle procedure by AA-I, no matter EGF was added in before, at same time or after AA-I stimulation.

CONCLUSIONAA-I (10 g L(-1)) has remarkable growth inhibition effects on survived RTEC, and can induce a blockage of G2/M phase in the cell cycle procedure. Exogenous EGF (20 microg L(-1)) promote the proliferation in normal cultured HK-2 cell. EGF treatment could not improve the proliferation inhibitory effect induced by AA-I, no matter adding EGF to cultures before, together with AA-I or after AA-I stimulation.

Aristolochic Acids ; pharmacology ; Cell Cycle ; drug effects ; Cell Line ; Cell Proliferation ; drug effects ; Epidermal Growth Factor ; pharmacology ; Epithelial Cells ; drug effects ; Flow Cytometry ; Humans ; Kidney Tubules, Proximal ; cytology ; drug effects ; Mutagens ; pharmacology

In the kidney, pericyte is the major source of myofibroblast (MyoF) in renal interstitium. It is reported that pericyte-myofibroblast transition（PMT）is one of the important pathomechanisms of renal interstitial fibrosis（RIF）. Among them, the main reasons for promoting RIF formation include pericyte recruitment, activation and isolation, as well as the lack of pericyte-derived erythropoietin. During the PMT startup process, pericyte activation and its separation from microvessels are controlled by multiple signal transduction pathways, such as transforming growth factor-β（TGF-β）pathway, vascular endothelial growth factor receptor (VEGFR) pathway and platelet derived growth factor receptor (PDGFR) pathway;Blocking of these signaling pathways can not only inhibit PMT, but also suppress renal capillaries reduction and further alleviate RIF. In clinic, many traditional Chinese medicine compound prescriptions, single traditional Chinese herbal medicine (CHM) and their extracts have the clear effects in alleviating RIF, and some of their intervention actions may be related to pericyte and its PMT. Therefore, the studies on PMT and its drug intervention will become the main development direction in the research field of anti-organ fibrosis by CHM.
Drugs, Chinese Herbal ; pharmacology ; Fibrosis ; Humans ; Kidney ; cytology ; drug effects ; pathology ; Myofibroblasts ; cytology ; Pericytes ; cytology ; Receptors, Platelet-Derived Growth Factor ; metabolism ; Signal Transduction ; Vascular Endothelial Growth Factor A ; metabolism