ABSTRACT
Objective:To investigate the regulation effect of endogenous nicotinamide phosphoribosyl transferase (Nampt) on the Vimentin expression of glomerular cells in high concentration glucose,and to clarify the mechanism of formation of diabetic kidney inflammation fibrosis.Methods:The C57/BL6 diabetic mice were selected and the kidney tissues were collected,and the wild C57/L86 mice were used as control group;the pathological section and tissue fluorescence staining were performed.The expression and location of endogenous Nampt and Vimentin in the glomerular cells were detected by immuno-focused technology.The HBZY-1 cells were randomly divided into 4 groups:low concentration of glucose (LG,0.56 mmol · L-1) control group,high concentration glucose (HG,200 mmol · L-1) group,HG +-FK866 group and HG+nicotinamide mononucleotide (NMN) group.In HG group,the cells were treated with FK866 (10 μmol · L-1) and NMN (1 mmol · L-1) for 24 h after cultured with HG for 5 d.The expression levels of Nampt,Vimentin,nuclear factor-kappa B p65 (NF-κBp65) and sirtuin type 1 (Sirt1) were detected by immunofluorescence and Western blotting methods.The expression levels of Nampt and Vimentin were detected by RT-PCR and Western blotting methods.Results:The shape and size of glomerulus had obvious atrophy of the mice in severe diabetic group compared with normal C57/BL6 mice group.The expression level of Vimentin in glomerular cells was increased with the increasing of endogenous Nampt (P＜0.01).When the HBZY-1 cells were cultured in HG condition,the exprssion levels of Nampt,Vimentin and NF-kB p65 were obviously increased while the Sirt1 expression levels was significantly decreased compared with control group (P＜ 0.01).The expression levels of Nampt,Vimentin and NF-κB p65 in glomerular cells in HG+FK866 HG+ NMN groups were singnificartyly decreased compared with control group (P＜0.01).Conclusion:The endogenous Nampt over-expression in glomerular cells can enhance the expression of Vimentin under high concentration of glucose stress through NF-κBp65 and Sirt1 signal pathway.
ABSTRACT
Objective:To investigate the regulation effect of endogenous nicotinamide phosphoribosyl transferase (Nampt) on the Vimentin expression of glomerular cells in high concentration glucose,and to clarify the mechanism of formation of diabetic kidney inflammation fibrosis.Methods:The C57/BL6 diabetic mice were selected and the kidney tissues were collected,and the wild C57/L86 mice were used as control group;the pathological section and tissue fluorescence staining were performed.The expression and location of endogenous Nampt and Vimentin in the glomerular cells were detected by immuno-focused technology.The HBZY-1 cells were randomly divided into 4 groups:low concentration of glucose (LG,0.56 mmol · L-1) control group,high concentration glucose (HG,200 mmol · L-1) group,HG +-FK866 group and HG+nicotinamide mononucleotide (NMN) group.In HG group,the cells were treated with FK866 (10 μmol · L-1) and NMN (1 mmol · L-1) for 24 h after cultured with HG for 5 d.The expression levels of Nampt,Vimentin,nuclear factor-kappa B p65 (NF-κBp65) and sirtuin type 1 (Sirt1) were detected by immunofluorescence and Western blotting methods.The expression levels of Nampt and Vimentin were detected by RT-PCR and Western blotting methods.Results:The shape and size of glomerulus had obvious atrophy of the mice in severe diabetic group compared with normal C57/BL6 mice group.The expression level of Vimentin in glomerular cells was increased with the increasing of endogenous Nampt (P＜0.01).When the HBZY-1 cells were cultured in HG condition,the exprssion levels of Nampt,Vimentin and NF-kB p65 were obviously increased while the Sirt1 expression levels was significantly decreased compared with control group (P＜ 0.01).The expression levels of Nampt,Vimentin and NF-κB p65 in glomerular cells in HG+FK866 HG+ NMN groups were singnificartyly decreased compared with control group (P＜0.01).Conclusion:The endogenous Nampt over-expression in glomerular cells can enhance the expression of Vimentin under high concentration of glucose stress through NF-κBp65 and Sirt1 signal pathway.
ABSTRACT
Objective To investigate the expression of nuclear factor-kappa Bp65 (NF-κBp65)and Toll-like receptor 4(TLR4)protein in the brain tissues of 7-day-old Sprague-Dawley(SD) rats with cerebral hypoxia-ischemia encephalopathy (HIE) and to explore the role of TLR4 and NF-κBp65 in the pathogenesis of neonatal rats with hypoxic-ischemic brain damage.Methods Seven-day SD rats were randomly divided into the experimental group and the control group.Brain pathological changes were observed in light microscopy at 6 h、12 h、24 h、72 h、7 d after HIE.The expression of TLR4 and NF-κBp65 in brain tissues were analyzed by immunohistochemistry method.Results NF-κBp65 and TLR4 were expressed in the neuron and microglia of control group and experimental group.The expression were most significant at cerebral cortex and hippocamp.However,the expression of NF-κBp65and TLR4 began to increase at HIE 6h:NF-κBp65 (0.219 3 ± 0.024 7,0.215 7 ±0.030 4)and TLR4(0.327 1 ±0.033 3,0.303 9 ±0.037 9),and achieved the hightest at HIE 24h:NF-κBp65 (0.3564±0.0235,0.3365 ±0.023 2)and TLR4(0.434 2 ±0.0428,0.4193 ±0.041 3),then decreased at HIE 72 h:NF-κBp65 (0.289 2 ± 0.032 0,0.260 9 ± 0.021 2) and TLR4 (0.300 5 ± 0.020 9,0.282 0 ± 0.022 6),and HIE 7 d:NF-κBp65(0.247 9 ±0.0340,0.242 1 ±0.025 4) and TLR4(0.274 4 ±0.0288,0.257 1 ±0.027 5).Conclusion There is a positive correlation between NF-κBp65 and TLR4 in rats with HIE.It suggested that they may have the same pathophysiology development in HIE.
ABSTRACT
This study was aimed to explore the effect of Catalpol on the cerebral infarction size in acute phase, water content and inflammatory reaction of early recovery after permanent middle cerebral artery occlusion (pM-CAO). Adult male Sprague-dawley rats were subjected to chemical method to establish MCAT model. The detec-tion was made on neurobehavioral symptoms, cerebral infarction volume and water content at 24 h after surgery. The content of intedeukin-6 (IL-6), intedeukin-10 (IL-10) and nuclear factor kappa Bp65 (NF-κBp65) were de-tected after pMCAO with enzyme-linked immunosorbent assay (ELISA). The results showed that 24 h after MCAT, Catalpol 15-60 mg·kg-1 can significantly improve the neurobehavioral symptoms (P < 0.01, or P <0.001). The Catalpol 15 mg·kg-1 can significantly reduce cerebral infarction volume (P < 0.05). The Catalpol 30-60 mg·kg-1 can significantly reduce water content (P < 0.05). Catalpol 30-60 mg·kg-1 can significantly improve neurobehav-ioral symptoms from the 7th day after pMCAO. On the 14th after pMCAO, the content of IL-10 and NF-κBp65 of ischemia brain had no difference compared with sham-operated group. The IL-6 level of ischemia brain was obvi-ously reduced than the sham-operated group(P < 0.05). The intragastric administration of Catalpol 15 mg·kg-1 for 14 days can reduce the content of NF-κBp65 in the ischemia brain of model rats (P < 0.05). It was concluded that Catalpol can improve neurobehavioral symptoms of acute and subacute phase after cerebral ischemia, reduce infarcts and water content. These effects may not be related with its inhibition of inflammatory derived from cere-bral ischemia.
ABSTRACT
Objective To explore the relationship of signal transduction among human papillomavirus 18 E6 oncoprotein (HPV18E6), signal transducers and activators of transcription 1 (STAT1), protein kinase R( PKR )/α subunit of eukaryotic initiation factor 2 ( eIF2α ), nuclear factor-kappa Bp65 ( NF-κBp65 ), mitogen-activated protein kinase( MAPK)/c-Jun N-terminal kinase(JNK) ,and possible molecular mechanism. Methods Construct two lentiviral vectors which contain shRNA interfering sequence aiming at the targets of HPV18E6 oncogene and NC sequence( HPV18E6-RNAi-LV, NC-GFP-LV), based on the transduction with HPV18E6-RNAi-LV and NC-GFP-LV into HeLa cell to interfere the expression of HPV18E6 oncogene and NC sequence,the expressions of mRNA and protein( including phosphating patem)of HPV18E6, STATI, PKR, eIF2α, NF-κBp65, MAPK, JNK are measured with RT-PCR and Western blot, the difference of proliferation and sensitivity to carboplatin of HeLa cell are determined with Transwell cell methods and MTT among every groups. Results The expression of HPV18E6 oncogene can affect the expression level of mRNA and protein of NF-κBp65 and PKR genes, also affect phosphating levels of phosphating protein p-STAT1, p-PKR and p-eIF2α;the restraining rates of proliferation and sensitivity to carboplatin of HeLa cell are higher in HPV18E6-RNAi-LV group than the other groups( P＜0. 05 or P＜0.01 ). Conclusion HPV18E6 oncoprotein not only reduces the expression of PKR but dephosphorylates p-STAT1, pPKR and p-eIF2α to restrain activation of PKR/eIF2α signal transduction passage, maintain the proliferation and invading ability of HeLa cell and restrain apoptosis. The signal transduction among HPV18E6, MAPK/JNK are not clear.