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Spinal cord injury can be divided into primary and secondary injury. As an important process of spinal cord injury, secondary injury can be classified into acute phase, subacute phase and chronic phase according to the time and progression of the injury. Oxidative stress reaction, inflammatory reaction, tissue edema, scar formation and other pathological changes appear subsequently during the process. In the central nervous system, the astrocyte is one of the most widely distributed cell that has different shapes at different stages and plays a complex role such as anti-inflammatory or pro-inflammatory action and neuroprotective or anti-neurorestorative effect after spinal cord injury. The astrocyte has been a research focus in the field of spinal cord injury. The authors review the role and research progress of astrocyte in oxidative stress response, excitotoxicity, inflammatory response, tissue edema, scar formation, axonal regeneration and cell transformation in spinal cord injury based on the pathological changes of secondary injury, in order to provide new ideas to the related research of spinal cord injury.
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Objective:To investigate the effects of tamoxifen on high glucose-induced epithelial-to-mesenchymal transition of rat peritoneal mesothelial cells and the underlying mechanism.Methods:The peritoneal mesothelial cells of normal male SD rats were selected between January 2015 and June 2016 and then cultured and divided into blank control, high-glucose stimulation and drug intervention groups. High-glucose stimulation group: primary cultured rat peritoneal mesothelial cells (RPMCs) were treated with 60 mmol/L high-concentration glucose to induce epithelial-to-mesenchymal transition. Drug intervention group: (1) RPMCs were treated with 60 mmol/L high-concentration glucose and different concentrations (0.5 μmol/L, 2 μmol/L) of tamoxifen. After 72 hours of stimulation, protein was extracted. (2) RPMCs were treated with 60 mmol/L high-concentration glucose and 2 μmol/L tamoxifen with or without 2 μmol/L ER-α antagonist for 1 hour to extract protein and for 6 hours to extract RNA. (3) RPMCs were treated with high-concentration glucose and 2 μmol/L tamoxifen with or without 1 μmol/L 1 μM proteasome inhibitor for 1 hour to extract protein. Western blot analysis was performed to analyze change in E-cadherin, α-SMA, Smad2, p-Smad2, Smad3, p-Smad3 and Smad4 protein. Real-time fluorescence quantitative PCR was performed to detect the change in mRNA expression of Smad2, Smad3, connective tissue growth factor and plasminogen activator inhibitor 1.Results:Tamoxifen attenuated epithelial-to-mesenchymal transition on RPMCs induced by high-level glucose, showing increased expression of epithelial cell marker E-cadherin and decreased expression of α-SMA in a concentration-dependent manner ( tE-cadherin = 2.31, tα-SMA =-2.53, both P < 0.05).TGF-β1/R-Smad signal pathway was activated by high-concentration glucose. Phosphorylation of Smad2/3 and mRNA expression of CTGF and PAI-1 were increased. Tamoxifen remarkably reduced protein and mRNA level of above mentioned protein and related target genes ( tp-Smad2 = -3.38, tCTGF = -3.81, P < 0.05), which could be blocked by ER-α antagonist. Finally, proteasome inhibitor could weaken the inhibitory effects of tamoxifen on p-Smad2/3 and increase p-Smad2/3 protein level ( tp-Smad2 = 3.94, P < 0.05). Conclusion:Tamoxifen activates ER-α on RPMCs, weakens the activation of TGF-β1/R-Smad signal pathway through decreasing p-Smad2 protein level, and effectively inhibits the progression of high-concentration glucose-induced epithelial-to-mesenchymal transition possibly through degrading p-Smad2 protein through proteasome. The role of tamoxifen in epithelial-to-mesenchymal transition may provide a possible guide for research, prevention and treatment of peritoneal fibrosis.
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Objective:To investigate the role and mechanism of (histone deacetylase 6, HDAC6) in the epithelial-mesenchymal transition (EMT) of renal tubular epithelial cells and the activation of renal interstitial fibroblasts.Methods:Human renal tubular epithelial cells (HK-2) and rat renal interstitial fibroblast (NRK-49F) were cultured in vitro, and divided into 4 groups: control group, Tubastatin A (TA) group (treated with 10 μmol/L HDAC6 inhibitor TA for 36 h), transforming growth factor-β1 (TGF-β1) group (10 ng/ml TGF-β1 for 36 h), and TGF-β1+TA group (treated with 10 ng/ml TGF-β1 and 10 μmol/L TA for 36 h). The expression levels of fibronectin, α-smooth muscle actin (α-SMA), collagen I, E-cadherin, HDAC6, acetyl histone H3, histone H3, acetyl α-tubulin, α-tubulin, TGF-β receptor (TGF-βR) 1, p-Smad3, Smad3, connective tissue growth factor (CTGF), epidermal growth factor receptor (EGFR) and p-EGFR in HK-2 and NRK-49F cell samples were detected by Western blotting, and quantitative analysis was performed according to gray level. Results:(1) In HK-2 cells stimulated by TGF-β1, TA decreased the expression of fibronectin, α-SMA, collagen I, and increased the expression of epithelial cell marker E-cadherin. Meanwhile, TA decreased the expression of HDAC6 and increased the expression levels of acetyl histone H3 and acetyl α-tubulin (all P<0.05). (2) Compared with the TGF-β1 group, the expressions of TGF-βR1, p-Smad3, CTGF and p-EGFR in TGF-β1+TA group were decreased (all P<0.05), while the total protein levels of Smad3 and EGFR were not significantly different (both P>0.05). (3) In NRK-49F cells stimulated by TGF-β1, TA decreased the expressions of fibronectin, α-SMA, collagen I, TGF-βR1 and p-Smad3 (all P<0.05). Conclusions:Blockade of HDAC6 by TA may inhibit the EMT of renal tubular epithelial cells and the activation of renal interstitial fibroblasts via regulating multiple signaling pathways including TGF-β/Smad3, CTGF and EGFR.
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Objective:To investigate the renal protective effect of Tangshenping capsule (Tangshenping) on diabetic nephropathy (DN) KKAy mice and its effect on Wnt/β-catenin signaling pathway. Method:Sixty female Sprague-Dawley KKAy mice aged 10 weeks old were induced with KKAy rat feed for 10 weeks. The DN animal model was successfully determined with blood glucose (>16.7 mmol·L-1) and 24 hour urine protein (>0.4 mg). The model mice were randomly divided into a model group, an irbesartan group, and low, medium and high-dose Tangshenping group, with 10 female C57BL/6J mice as a control group. The treatment groups were given the corresponding drugs by gavage. The normal group and the model group were given an equal volume of deionized water by gavage. The intragastric dose was 0.01 mL·g-1 body weight coefficient once a day. The general conditions of the mice were observed, the body mass was weighed every 4 weeks, and 24 h urine protein was quantified. At the 26th week, the blood was collected from eyeballs, and the mice were put to death. The quality of the kidneys, serum blood urea nitrogen (BUN), serum creatinine (SCr), triglyceride (TG), malondialdehyde (MDA), nitric oxide (NO) and superoxide dismutase (SOD) content were measured. In situ hybridization and immunohistochemistry were used to detect the expressions of Wnt4, glycogen synthase kinase 3β(GSK3β) and β-catenin in kidney tissues. Result:Compared with model group, body mass, kidney mass/body mass, and 24 h urine protein were significantly lower in high-dose Tangshenping group (PPPβ and β-catenin were decreased (PConclusion:Tangshenping may inhibit the activation of Wnt/β-catenin signaling pathway, reverse the transdifferentiation of renal tubular epithelial cells in DN KKAy mice, delay the progression of renal interstitial fibrosis, and then exert renal protection.
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Traumatic brain injury (TBI) is a central nervous system disease with increasing incidence,morbidity and mortality worldwide.TBI can affect the integrity of neuron,causing neuronal axons damage or death of neurons,which results in serious sequelae.After TBI,astrocytes (AST) in the cerebral cortex will be activated into reactive astrocytes (RAS).RAS in the early stage of TBI has a certain repair effect on the injury.However,RAS will proliferate to form glial scars,which has adverse effects on nerve function repair after injury.Therefore,controlling the status of RAS is the key to the treatment of TBI.In recent years,it has been proved that RAS can be transdifferentiated into neurons by transdifferentiation technology,which can not only remove glial scars,but also integrate with the microenvironment at the injury site to replace the injured neurons,which is of great significance for the repair of nerve function after TBI.This article reviews the types of transdifferentiation and the different pathways of RAS transdifferentiation into neurons,aiming to have a better understanding of the research progress of RAS transdifferentiation into neurons to repair TBI.
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In regenerative medicine, growing cells or tissues in the laboratory is necessary when damaged cells can not heal by themselves. Acquisition of the required cells from the patient's own cells or tissues is an ideal option without additive side effects. In this context, cell reprogramming methods, including the use of induced pluripotent stem cells (iPSCs) and trans-differentiation, have been widely studied in regenerative research. Both approaches have advantages and disadvantages, and the possibility of de-differentiation because of the epigenetic memory of iPSCs has strengthened the need for controlling the epigenetic background for successful cell reprogramming. Therefore, interest in epigenetics has increased in the field of regenerative medicine. Herein, we outline in detail the cell trans-differentiation method using epigenetic modification for bone regeneration in comparison to the use of iPSCs.
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Bone Regeneration , Cell Transdifferentiation , Cellular Reprogramming , Epigenomics , Induced Pluripotent Stem Cells , Memory , Methods , Regenerative Medicine , Tissue EngineeringABSTRACT
Objective To investigate the effect of lysine methyltransferase SET8 on regulating cell transdifferentiation of rat vascular smooth muscle cells(VSMCs)into osteoblast-like cells. Methods VSMCs were obtained from rat thoracic aorta,and then randomly divided into control group(non-transfection),the empty plasmid group(transfect NS-shRNA)and SET8-shRNA group. The expression of SET8, runt-related transcription factor 2 (RUNX2) was detected by RT-PCR and Western blot assay. Alkaline phosphatase (ALP) activity was measured by enzyme linked immunoassay. Results The expression of SET8 in VSMCs was effectively inhibited by SET8-shRNA.RT-PCR and Western blot results showed that the expression of RUNX2 was decreased in cells after SET8-shRNA transfection (P<0.05). ALP activity was significantly reduced after SET8-shRNA transfection(P<0.05).Conclusion Interference with SET8 gene expression could inhibit the differentiation of VSMCs into osteo-like cells.
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Objective To explore the impacts of NPHP1 knockdown on the phenotype of Madin-Darby canine kidney (MDCK) cells. Methods The expression of NPHP1 in MDCK cells was knockdown by siRNA interference. Cells were divided into normal control group, negative control group and siRNA group. The cellular morphology and migration were observed by light microscope. The mRNA expressions and activities of matrix metalloproteinases 2 and 9 (MMP2 and MMP9) were detected by real time PCR and gelatin zymography. The mRNA and protein expressions of E-cadherin, β-catenin, zonula occluden-1 (ZO-1), ZO-1-associated nucleic acid binding protein (ZONAB) and α-smooth muscle actin (α-SMA) were analyzed by real time PCR, Western blotting and immunocytochemistry. Results Compared with those in normal control group, in siRNA group the mRNA expressions of E-cadherin, β-catenin and ZO-1 decreased, and MMP9, MMP2, α-SMA and ZONAB increased after interfering NPHP124 h (all P<0.05); the protein expressions of E-cadherin,β-catenin and ZO-1 decreased and ZONAB and α-SMA increased after 48 h (all P<0.05), and MDCK cells became elongated with enhanced migration capacity; siRNA cells had decreased expressions of E-cadherin and β-catenin on the membrane, but increased expression of ZONAB in cytoplasm and nucleoplasm after 72 h, and α-SMA was also observed in some interfered cells. Conclusions NPHP1 knockdown induces epithelial-mesenchymal transition in MDCK cells, and ZO-1/ZONAB signaling pathway was activated. These changes may associate with renal interstitial fibrosis of Nephronophthisis type I.
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Objective To investigate the role of STAT3 transcription factor in IL-6 inducing epithelial mesenchymal transition (EMT) of human peritoneal mesothelial cells (HPMCs).Methods HPMCs were cultured in vitro and grouped.(1) According to the stimulation time with 50 μg/L IL-6,HPMCs were divided into 24,48,72 h groups.(2) HPMCs were grouped 50,100 μg/L according to IL-6 concentration.(3) HPMCs were respectively divided into control group,IL-6 group,empty vector group,empty vector+IL-6 group,virus infecting group and virus infecting+IL-6 group,as lenti-virus vector mediating RNA interference targeting STAT3 was applied.The mRNA expressions of E-cadherin,α-smooth muscle actin (α-SMA) and vascular endothelial growth factor (VEGF) were detected by real time PCR;their protein expressions and the phosphorylation of JAK2 and STAT3 were detected by Western blotting;the expressions and distribution of E-cadherin and α-SMA were observed by immunofluorescence.Results Compared with those in control group,the expression of E-cadherin decreased remarkably (P ＜ 0.05),while the expressions of VEGF and α-SMA and the ratio of phosphorylated (p)-JAK2/JAK2 and p-STAT3/STAT3 increased significantly in IL-6 concentration groups and stimulation time groups (all P ＜ 0.05),which had been dose and time dependent.Compared with empty vector+IL-6 group,virus infecting+IL-6 group had decreased expressions of VEGF and α-SMA,while increased expressions of E-cadherin (all P ＜ 0.05).Conclusions IL-6 can promote VEGF and α-SMA gene expression and prevent E-cadherin gene expression by STAT3,which involves in EMT of peritoneum fibrosis.While STAT3 gene is knocked-down,EMT is inhibited in HPMCs.
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Objective To investigate the role of STAT3 transcription factor in IL-6 inducing epithelial mesenchymal transition (EMT) of human peritoneal mesothelial cells (HPMCs).Methods HPMCs were cultured in vitro and grouped.(1) According to the stimulation time with 50 μg/L IL-6,HPMCs were divided into 24,48,72 h groups.(2) HPMCs were grouped 50,100 μg/L according to IL-6 concentration.(3) HPMCs were respectively divided into control group,IL-6 group,empty vector group,empty vector+IL-6 group,virus infecting group and virus infecting+IL-6 group,as lenti-virus vector mediating RNA interference targeting STAT3 was applied.The mRNA expressions of E-cadherin,α-smooth muscle actin (α-SMA) and vascular endothelial growth factor (VEGF) were detected by real time PCR;their protein expressions and the phosphorylation of JAK2 and STAT3 were detected by Western blotting;the expressions and distribution of E-cadherin and α-SMA were observed by immunofluorescence.Results Compared with those in control group,the expression of E-cadherin decreased remarkably (P ＜ 0.05),while the expressions of VEGF and α-SMA and the ratio of phosphorylated (p)-JAK2/JAK2 and p-STAT3/STAT3 increased significantly in IL-6 concentration groups and stimulation time groups (all P ＜ 0.05),which had been dose and time dependent.Compared with empty vector+IL-6 group,virus infecting+IL-6 group had decreased expressions of VEGF and α-SMA,while increased expressions of E-cadherin (all P ＜ 0.05).Conclusions IL-6 can promote VEGF and α-SMA gene expression and prevent E-cadherin gene expression by STAT3,which involves in EMT of peritoneum fibrosis.While STAT3 gene is knocked-down,EMT is inhibited in HPMCs.
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Objective To explore the role and mechanism of microRNA-15b in the regulation of epithelial-mesenchymal transition (EMT) of human peritoneal mesothelial cells (HPMCs).Methods PCR assay was used to determine the expression of microRNA-15b in the HMrSV5 induced by 138mmol/L high glucose for 24 h.MicmRNA-15b mimic or inhibitor was transfected into human peritoneal mesothelial cells (HMrSV5) to over-express or down-regulate microRNA-15b.The cells were then incubated with 138 mmol/L high glucose for 24 h,and the expressions of E-cadherin(E-Cad),Vimentin (VIM),Fibronectin(FN) and Smad7 were detected by real-time PCR and Western blotting respectively.Results microRNA-15b in the HMrSV5 ceils was over-expressed and down-regulated.Increased level of microRNA-15b was obtained in HMrSV5 cells treated with high glucose.In vitro,high glucose led to the up-regulation of vimentin as well as fibronectin and the down-regulation of E-cadherin in HMrSV5 cells (all P ＜ 0.05),which indicated EMT and fibrosis.Suppression of microRNA-15b by transfection with microRNA-15b inhibitor partially reversed the EMT and fibrosis changes (P ＜ 0.05),while over-expression of microRNA-15b by transfection with microRNA-15b mimic obviously enhanced the EMT and fibrosis changes (P ＜ 0.05).Conclusions MicroRNA-15b mediates high glucose induced EMT in human peritoneal mesothelial cells by the inhibition of Smad7 possibly.MicroRNA-15b maybe a new target for the prevention and treatment of peritoneal fibrosis during peritoneal dialysis (PD).
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Objective To explore the effect of endoplasmic reticulum stress (ER stress) in uric acid?induced phenotypic change in renal tubular epithelial cells (HK?2). Methods (1) HK?2 cells were cultured with 0, 75, 150, 225, 300 mg/L uric acid for 24 h in vitro. (2) The cells were divided into normal control group, ER stress inhibitor 4?PBA (5 μmol/L) group, uric acid (150 mg/L) group and 4?PBA+uric acid group for 24 h. Morphological changes of HK?2 cells were observed under inverted microscope. MTT assay was used to detect the proliferation of HK?2 cells treated with 150 mg/L uric acid for 24, 48 and 72 h. The protein expressions of α?smooth muscle actin (α?SMA), vimentin, snail, glucose regulated protein 78 (GRP78) and the phosphorylation of eukaryotic initiation factor 2α(p?eIF2α) in HK?2 cells were measured by Western blotting. Results Compared with the control group, HK?2 cells in uric acid groups (150, 225, 300 mg/L) showed fibroblast?like appearance. The protein expressions of α?SMA, vimentin, snail, GRP78 and p?eIF2α in 150 mg/L and 225 mg/L uric acid groups were higher than those in the control group (all P<0.05). The proliferation of HK?2 cells in 150 mg/L uric acid group was lower than that in control group at 48 and 72 h (all P<0.01). Compared with the uric acid group, the cell morphology in 4?PBA+uric acid group was improved, and the protein expressions ofα?SMA, vimentin, snail, GRP78 and p?eIF2α were decreased (all P<0.05). Conclusions Uric acid may induce the phenotype transformation of renal tubular epithelial cell, and ER stress is involved. 4?PBA may inhibit the uric acid?induced ER stress response and phenotypic transformation, and may be beneficial in attenuating uric acid?induced renal tubular damage.
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Objective To observe the epithelial mesenchymal transition (EMT) of podocyte induced by high glucose,and to explore the potential protective mechanism of ursolic acid (UA).Methods The podocytes cultured in vitro were divided into four groups:normal group (glucose 5.5 mmol/L),mannitol group (glucose 5.5 mmol/L+mannitol 19.5 mmol/L),high glucose group (glucose 25 mmol/L) and UA group (glucose 25 mmol/L + UA 5 μmol/L).Podocyte morphology changes were observed by inverted phase contract microscope.The expression of zonula occludens-1 (ZO-1) and α-smooth muscle actin (α-SMA) were detected by immunofluorescence.The expressions of β-catenin and glycogen synthesis kinase-3β (GSK3β) were detected by Western blotting.The expressions of Wnt1,Wnt3a,Wnt5a,Wnt5b and GSK3β were detected by real-time PCR.Results Podocytes showed irregular arborization shape in normal glucose and transited to longer cobblestone-like shape as mesenchyme cell by high glucose culture.Compared with normal group,the expression of ZO-1 protein was down-regulated and the expression of α-SMA was up-regulated by high glucose culture (P ＜ 0.05).The expression of Wnt5a mRNA was down-regulated;β-catenin mRNA and protein were up-regulated (P ＜ 0.05);and GSK3β protein was down-regulated by high glucose culture (P ＜ 0.05).Compared with high glucose group,ursolic acid inhibited podocyte EMT,up-regulated the expression of ZO-1 protein,Wnt5a mRNA,GSK3β (P ＜ 0.05),and down-regulated the expressions of α-SMA protein,β-catenin mRNA and protein (P ＜ 0.05).Conclusion Ursolic acid attenuates high glucose induced epithelial mesenchymal transition of podocyte by inhibiting Wnt/β-catenin signaling pathway.
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Objective To explore the effect and the possible pathway of different concentrations of QLT0267,which was the inhibitor of the integrin-linked kinase (ILK),on the process of high glucose-induced tubularepithelial-myofibroblast transdifferentiation (TEMT) in human renal tubular epithelial cells (HK-2).Methods HK-2 cells were exposed to 30 mmol/L GS,and TEMT model was established.After excluding the effect of high osmotic in TEMT,HK-2 cells were divided into 6 groups by different concentrations of GS and QLT0267 for 48 hours.The rate of the cell proliferation was calculated by MTT.The expression of ILK and α-smooth muscle actin (α-SMA) were determined by immunofluorescence and Western blot,and the expression of protein kinase B (AKT),phosphorylated protein kinase B (p-AKT),and E-cadherin were determined by Western blot.Results (1) The expression of ILK,p-AKT,and α-SMA in HK-2 cells were unregulated and the expression of E-cadherin was downregulated for 48 hours with glucose treating vs control (P ＜ 0.05);(2) The proliferation rate in high glucose group was higher than the group which concentration of QLT0267 was greater than 5 μmol/L (P ＜ 0.05);(3) With the concentrations of QLT0267 increased,the expression of p-AKT,α-SMA was gradually decreased (all P ＜ 0.05),and the expression of E-cadherin was gradually increased (all P ＜ 0.05).Conclusions 30 μmol/L of GS can lead to TEMT in HK-2 cell.The QLT0267 with concentration greater than 5 μmol/L may prevent the activation of ILK downstream proteins,then partially inhibits cell proliferation and TEMT in HK-2 cell.
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Objective To observe the effect of JLP on transdifferentiation of human renal proximal tubular epithelial cells (HK-2),and to investigate the role of p38 MAPK signaling pathway in this process.Methods The knock-down plasmids of JLP were constructed.HK-2 cells were randomly divided into four groups:negative control cells (Ctrl-shRNA group),knock-down jlp cells (jlpshRNA group),negative control cells with FGF-2 treatment (FGF-2 group) and knock-down jlp cells with FGF-2 treatment(jlp-shRNA +FGF-2 group).The expressions of JLP,E-cadherin,TGF-β1,α-SMA,p-p38 MAPK protein were detected by Western blotting.After the induction of FGF-2 for 24 hours,the expressions of α-SMA,COL-Ⅰ,FN were detected by immunocytochemistry.Results Compared with Ctrl-shRNA group,the expression of JLP protein was significantly down-regulated in FGF-2 group.Compared with FGF-2 group,the expressions of TGF-β1,α-SMA,p-p38 MAPK protein were significantly up-regulated,while E-cadherin protein was significantly down-regulated (P ＜ 0.05).Compared with FGF-2 group,the expressions of α-SMA,COL-Ⅰ,FN immunostaining increased markedly in jlp-shRNA+FGF-2 group.Conclusion Scaffolding protein JLP is critical in preventing EMT in the course of fibrosis through the inhibition of p-p38 activation in HK-2 cells.
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Objective To explore the effect of irbesartan on cardiac endothelial-mesenchymal transition (EndMT) in diabetic rats.Methods The model of diabetic rat was induced by intraperitoneal injection with streptozotocin (STZ,35 mg/kg) in spontaneous hypertensive rats (SHR).Diabetic rats were divided into diabetic group and the Irbesartan treated group.The pathological changes were investigated by fluorescence microscope and electron microscope.The EndMT was studied in human aortic endothelial cells (HAEC) exposure to high glucose.The concentration of angiotensin Ⅱ in the supernatant was detected by radioimmunoassay.Immunofluorescence staining was performed to detect the co-localization of CD31 and FSP1.Results The significant myocardial fibrosis was presented in the diabetic group.Endothelial protrusions were prominent feature in myocardial microvascular of diabetic rat compared with the control group rats.Double staining of HAEC showed co-localization of CD31 and FSP1,which was decreased by the treatment of Irbesartan (P ＜ 0.05).When HAEC was exposed to high glucose,it showed some cells acquired spindle-shaped morphology and lost CD31 staining,and FSP1 and α-SMA protein expression levels were markedly upregulated,which attenuated by the treatment of Irbesartan.Conclusion Irbesartan might prevent diabetes from myocardial fibrosis via inhibition of EndMT in diabetic rats.
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Objective To investigate the role of microRNA-129-5p (miR-129-5p) in the regulation of epithelial-mesenchymal transition (EMT) of human peritoneal mesothelial cells (HPMCs) isolated from peritoneal dialysate effluents and TGF-β1 induced HPMCs line.Methods The isolated cells were cultured from peritoneal dialysate effluents overnight of 10 patients just started PD and 12 patients with PD over 6 months.Taqman PCR assay was used to determine the expression of miR-129-5p in the HPMCs.Moreover,the expression of miR-129-5p in HPMCs induced by 5 μg/L TGF-β1 for 0-72 h was also detected by Taqman PCR.HPMCs were pre-transfected with miR-129-5p precursor (pre-mir-129-5p) to overexpress miR-129-5p,then incubated with TGF-β1 for 48 h,and the expression of EMT associated gene and protein was detected by real-time PCR,Western blotting and immunofluorescence,respectively.Furthermore,the effect of TGF-β1 on the expression of Smad interacting protein-1 (SIP1) and the regulation of pre-miR-129-5p on the SIP1 expression also were investigated.Results MiR-129-5p expression significantly down-regulated in the HPMCs isolated from PD patients over 6 months than from PD start patients(P ＜ 0.01).Similarly,TGF-β1 remarkably decreased miR-129-5p in HPMCs lines on time-dependent manner (P ＜ 0.01).Pre-mir-129-5p dramatically restored the expression of epithelial marker E-cadherin,while inhibited the expression of Vimentin,a mesenchymal marker,in HPMCs induced by TGF-β1 (all P ＜ 0.01).In addition,TGF-β1 increased SIP1 expression in HPMCs time dependently,while the high level of SIP1 protein was obviously repressed after transfected of pre-miR-129-5p (P ＜ 0.01),but there was no obvious change of its mRNA expression.Conclusion MiR-129-5p modulates EMT formation of HPMCs in PD process,possibly by posttranscriptional inhibition of SIP1.Targeting miR-129-5p/SIP1 may provide a new approach for the prevention and treatment of peritoneal fibrosis during PD.
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Objective To study the action of GSK-3βin podocyte transdifferentiation effecundethe high glucose condition . MethodPodocytewere treated in RPMI-1640 medium with differenconcentrationof glucose fo36 h .The expressionof neph-rin ,podocin ,α-Smand Fibronectin were detected by both Western bloand indirecimmunofluorescence analysi.Athe same time ,the change of albumin inflow amounof podocytein differentreatmengroupwadetected by using the Transwell cham-be.The changeof expression amounand activity of GSK-3βin high glucose condition were detected by using the GSK-3 activity assay ki.The phenotypiand functional changeof podocytewere detected aftedisturbing GSK-3βby GSK-3βsiRNin the high glucose group .ResultThe expression levelof nephrin and podocin protein were down-regulated with the increase of the glucose concentration in dose-dependenmanner(P＜0 .05) ,and the expression level of α-Smwaup-regulated with the increase of the glucose concentration in dose-dependenmanner(P＜0 .05);the albumin inflow amounof podocytewaup-regulated with the in-crease of the glucose concentration in dose-dependenmanner(P＜0 .05) .The expression amounof GSK-3βin the high glucose group waincreased(P＜0 .05) .Compared with the control group ,the expression amounof nephrin and podocin aftehigh glucose treatmenin the GSK-3β siRNtreatmengroup waincreased ,while the expression of α-Smwadecreased .Conclusion The high glucose condition could induce the phenotypichange and functional impairmenof mice podocyte;GSK-3βparticipatein the epithelial-mesenchymal transdifferentiation procesof podocyte undethe high glucose condition .
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A metaplastic papillary tumor of the Fallopian tube is an extremely uncommon condition, with odd and confusing features that make it difficult to categorize as benign or borderline. Here, we summarize all the published cases to date and document the case of a 41-year-old woman diagnosed with this alteration after her last childbirth and ensuing tubal ligation. One of the tubes was bulky and filled with a caramel-like substance encircling a blurry spot. Light microscopy detailed a slender stalk covered by eosinophilic, columnar plump cells, showing atypical nuclei and focal budding. Mitotic figures were absent. The immunohistochemistry panel was positive for pan-cytokeratin, epithelial membrane antigen, cyclin D1, and hormone receptors. Additionally, a proliferation index of less than 5% was rated using Ki-67. The true nature of this tumor (reactive vs neoplastic) is uncertain. Nonetheless, its association with pregnancy suggests an adaptive change, likely similar to the atypical transdifferentiation proposed for Arias-Stella reaction.
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Adult , Female , Humans , Pregnancy , Cell Transdifferentiation , Cyclin D1 , Eosinophils , Epithelium , Fallopian Tubes , Immunohistochemistry , Microscopy , Mucin-1 , Parturition , Sterilization, TubalABSTRACT
Melanoma arises from the melanocyte cell lineage, it is one of the most notoriously aggressive and treatment-resistant human cancers. Microphthalmia-associated transcription factor (MITF) acts as a master regulator of melanocyte's development, function and survival by modulating various differentiation and cell-cycle progression genes. It also plays an oncogenic role in melanoma. MITF targeting genes important in melanoma progression include promoting survival and blocking apoptosis, stimulating proliferation, and inhibiting cell cycle. This review summarized the molecular function of MITF and its related pathways, and hope it will shed light on strategies for improving therapeutic approaches for melanoma. Copyright © 2013 by TUMOR.