Effects Of Parthenolide And Its Derivatives On High Glucose-induced Cell Proliferation, NF-κB Activation And MCP-1Expression In Rat Mesangial Cells

With the development of modern society and people living standard rise, and Diabetes (Diabetes mellitus, DM) incidence has risen dramatically throughout the world. Diabetic nephropathy (Diabetic nephropathy, DN) is the DM patients with one of the most common and most serious microvascular complications, is the most major cause of end-stage renal disease. At present, the pathogenesis of DN is not fully clear, still lack of effective treatment methods. Therefore, how to effectively prevent and cure DN has become a problem in clinical. The cause of DN has not been fully elucidated, now thought to be caused by a variety of causes. Morbidity associated with genetic, autoimmune, and environment. DM is popular all over the world of a kind of metabolic disease, the DN is the most common complication of diabetes in the west has become the main cause of the chronic renal failure. Glomerular mesangial cell proliferation and cytokine secretion and the accumulation of extracellular matrix plays an important role in the DN glomerular sclerosisHyperglycemia is a sign of DN patients sugar metabolic disorder. DN is the basic pathological changes of glomerulus vascular dysfunction and the accumulation of extracellular matrix, and lead to thickening of glomerular basement membrane and glomerular sclerosis and disfunction. Proinflammatory factors stimulation result in mesangial cell proliferation and secretion of the accumulation of extracellular matrix and many proinflammatory factor, so the glomerular mesangial cells are target cells of diabetic nephropathy, inflammation is the pathogenesis of diabetes and its complications. DN may seriously affect the national health service, and to bring the state and society a heavy economic burden and social burden, therefore, seeks to etiology and pathogenesis of DN, explore new effective treatment measures is imminent.Hyperglycemia is the pathogenesis of DN initiating and the center link, strict control of blood sugar can reduce and delay the occurrence and development of DN. Nuclear factor KappaB (NF-κB) is a complex and widely distributed in many cell types of transcription factors, glomerular mesangial cells, is no exception. The nf-kappa B for signal transmission and gene expression plays an important role in the process of inflammation reaction. Affected by hyperglycemia NF-κB is activated, it is formed many inflammatory factors such as the cause of the adhesion molecules and pro-inflammatory cytokines, including inflammatory cell factor Nuclear factor kappa B predominate-(Nuclear factor-kappa B, NF-κB) and Monocyte chemotactic protein1(Monocyte chemoattractant protein-1, MCP-1) play an important mediating role. I kappa alpha B is transcription factors predominate the NF-κB inhibitor, it can control immune and inflammatory response. The NF-κB because IκBα predominate in the presence of protein in the cytoplasm in the stationary state, many extracellular immune stimulation can induce the IκKβ predominate activated receptor mediated, by cause IκKβ predominate phosphorylation and degradation, and cause the NF-κB is activated. Activate the NF-κB into the nucleus and promote the release of proinflammatory cytokines. Excessive expression of pro-inflammatory factor is another important factor which causes the DN. PTL, therefore, research on sugar induced rat glomerular MC NF-κB activity and inflammatory factors of MCP-1expression of inhibitory effect, in the treatment of DN may play an important role in the process, the delay may have important clinical significance for the development of DN, but the mechanism remains to be further research.Clinical treatment of diabetic nephropathy at present is mainly by control high blood sugar, high blood pressure, reduce proteinuria, protein restriction and correct the lipid metabolism disorders and anti-inflammatory measures such as symptomatic treatment, can still hasn’t found completely blocking drugs to the further development of DN. Therefore, actively explore the effective measures of treating DN, to prevent or delay the development of DN has important significance. Feverfew lactone parthenolide) is extracted from compositae plants out of the sesquiterpene lactone compounds, the folk have been widely used for fever, arthritis and migraine headaches and other inflammatory diseases. This experiment on rat glomerular mesangial cells as the object of experimental research, observation of feverfew lactones of high sugar nf-kappa B mesangial cells induced inhibition of active and MCP1expression, discusses feverfew lactones of diabetic nephropathy may produce protective effect, so as to provide theoretical and experimental basis for its clinical application.Chart Ⅰ Effects of High Glucose on Rat Glomerular Mesangial Cells Proliferation, MCP-1Expression and NF-κB ActivationObjective To explore the effect of high glucose on cells proliferation, monocyte chemoattractant protein-1(MCP-1)expression and NF-κB activation in rat mesangial cell(MC).Methods The in vitro culture of rat glomerular mesangial cells were divided into three groups:normal glucose group (glucose concentration of5.6mmol/L), high glucose1group (glucose concentration of15mmol/L), high glucose2group (glucose concentration of20mmol/L), high glucose3group (glucose concentration of 25mmol/L). MTT method was used to detect cell proliferation. MCP-1in the supernatant was measured by ELISA. IκBα expression was detected by Western Blot method to reflect the NF-κB activation.Results The MC proliferation and MCP-1expression were significantly increased after24h in high glucose groups. The expression of NF-κB-binding protein IκBα was obviously declined in high glucose30min later, which indicates that high glucose can induce the NF-κB activation in MCs.Conclusion High glucose-stimulated rat glomerular mesangial cell proliferation and promoted inflammation factor MCP-1expression may be through the NF-κB activation to achieve.Chart Ⅱ Effects of Parthenolide on high glucose-induced cell proliferation, NF-κB activation and MCP-1expression in rat mesangial cellsObjective To explore the effects of parthenolide (PTL) on high glucose-stimulated cell proliferation, NF-κB activation and monocyte chemoattractant protein-1(MCP-1)expression in rat mesangial cells(MCs).Methods MCs were cultured in normal glucose (glucose concentration of5.6mmol/L), high glucose (glucose concentration of30mmol/L) and high glucose with PTL respectively, MTT method was used to detect cell proliferation, MCP-1in the supernatant was measured by ELISA, the IκBα was detected by Western Blot method to reflect the NF-κB activation, EMSA method was used to measure the activation of NF-κB.Results The MC proliferation, MCP-1expression and NF-κB activation were significantly enhanced obviously and NF-KB-binding Protein IκBα obviously declined in high glucose; MC proliferation, MCP-1expression and the NF-κB activity were significantly abolished, I κ B α degradation was significantly reduced under the application of PTL.Conclusion PTL can restrain high glucose-stimulated NF-κB activation and MCP-1expression in rat MC, and provide a theoretical basis for its clinical application in the prevention and control of diabetic nephropathy.Chart Ⅲ Parthenolide and its derivatives inhibit highglucose-induced NF-κB activation and MCP-1expression in rat mesangial cellsObjective To explore the effects of independent synthetic parthenolide (PTL) and its derivatives MCL, LJ-5-42, ZJD-3-35a, ZJD-4-26a, CY-5, ZJD-3-35b, ZJD-3-11d, ZJD-2-24and ZJD-4-20on the high glucose-stimulated NF-κB activation and monocyte chemoattractant protein-1expression in rat MCs.Methods MC cells were cultured with50mmol/L glucose with MCL, LJ-5-42, ZJD-3-35a, ZJD-4-26a, CY-5, ZJD-3-35b, ZJD-3-lld, ZJD-2-24and ZJD-4-20respectively, MTT method was used to detect cell proliferation, enzyme-linked immunosorbent (ELISA) analysis was performed to determine the expressions of monocyte chemoattractant protein (MCP)-1in the supernatant, Real-time quantitative PCR method was used to detect MCP-1mRNA, Western Blot method was used to detect IκBα protein, NF-κB activity was evaluated by electrophoretic mobility shift assay.Results MC proliferation, MCP-1expression, the NF-κB activity enhanced obviously and the NF-κB binding protein I κ B α was obviously degradation in high glucose-stimulated MCs; MC proliferation, MCP-1expression and the NF-κB activity were significantly abolished, I κ B a degradation was significantly reduced under the application of PTL and its derivatives MCL, LJ-5-42, ZJD-3-35a, ZJD-4-26a and CY-5.Conclusion PTN and its derivatives MCL, LJ-5-42, ZJD-3-35a, ZJD-4-26a and CY-5can effectively attenuate the high glucose-stimulated NF-κB activation, IκBα degradation and monocyte chemoattractant protein-1expression in rat MCs, this may have an important role to the prevention and control of DN, may have important clinical significance to delay the occurrence of DN development, but the exact effect and its mechanism remains to be further research.