The Protection On Doparminergic Nerve Cells Exerted By Curcumin Via Inhibiting RAGE/NF-?B Pathway Of BV2 Cells

Background and objectiveParkinson's disease?Parkinson diseases,PD?is the alzheimer's disease?Al zheimer's diseases,AD?after the elderly,the second largest popular neurodegenerative diseases,the prevalence increased with the increase of age.Its main pathological feature is the midbrain substantia nigra compacta?SN-PC?Dopamine neurons?Dopamine,DA?loss and the formation of inclusion body in neuronal cytoplasm.Because of the substantia nigra polyamine can loss of neurons,result in striatal dopamine neurotransmitter levels drop,the clinical symptoms appear when the number of neurons achieves to 70%-more than 80%.Its motion characteristic symptoms mainly contain static tremor,muscle rigidity,bradykinesia,pose obstacles.The not motion symptoms including hyposmia,sleep disorders,autonomic nerve dysfunction,mental disorders?depression or anxiety?.The present study suggests that environmental factors,genetic factors,and central nervous system related to many factors such as interaction of ageing is related to Parkinson.Specific pathogenesis is related to oxidative stress,mitochondrial dysfunction,calcium ion steady-state misadjustment,Proteasome dysfunction,inflammation,immune response,excitability toxicity,apoptosis,etc,but the specific mechanism of PD is not entirely clear,need further study.Curcumin?Cur?is an effective component of turmeric,it is a kind of polyphenols extracted from the rhizome of turmeric gold plants,formula for C₂₁H₂₀O₆,relative molecular mass of 368.89.Color is orange yellow crystalline powder,insoluble in water and ether.Soluble in alcohol and alkaline solution,reddish-brown in alkaline,in neutral,acid is light yellow.Often used as food colorants,ph indicator,meat food spices and herbs.Curcumin has anti-oxidant,anti-inflammatory,anticoagulation,fall fat,cholesterol,anti-atherosclerosis,anti-aging,eliminating free radical and inhibiting tumor growth,cholagogic and antibacterial effect,low toxicity,less adverse reaction.Studies had shown that curcumin by depressing nuclear factor kappa B?nf-kappa B?predominate and inflammatory mediators such as Intercellular adhesion molecule-1?ICAM-1?,tumor necrosis factor alpha?TNF alpha?,interleukin 6?IL-6?,interleukin 1??IL-1??and iNOS effect into full play.RAGE is immunoglobulin superfamily receptors,are widely expressed in multiple,including neurons and microglia cells,and combined with many kinds of ligands to produce a series of biological effects.,for example,advanced glycosylation product.The nf-kappa B is extracted from B cell nuclear transcription factor,is an important transcription regulatory factor,also is the key of the inflammatory response conduction factor.The RAGE receptor activation can awaken the resting state nf-kappa B,the nf-kappa B to further adjust the expression of its downstream factor.The nf-kappa B involved in cell proliferation,apoptosis,inflammatory reaction,immune and other important process.AGEs combined with RAGE specific cell surface receptors to activate the nf-kappa B,the nf-kappa B stimulate a variety of inflammatory related cytokines,such as ICAM-1,TNF alpha,IL-6,IL-1?,the formation of iNOS and release.Curcumin maybe though inhibiting AGEs combined with the specificity of the cell surface receptor RAGE and suppressing the nf-kappa B activation of nuclear transcription factor,the releasing of inflammatory factors plays a protective role in the dopamine neurons.This study choose curcumin treatment BV2 cells induced by rotenone model,collecting BV2 cells supernatant as SH-SY5Y cells conditioned medium processed SH-SY5Y cells.Detect BV2,SH-SY5Y cell vitality,BV2 cells RAGE and nf-kappa B protein expression,BV2 cell culture supernatant cytokine levels,conditioned medium influence on SH-SY5Y cells proliferation activity.Discussion the protective effects of curcumin,to dopamine neurons and the relationship among curcumin,RAGE,nf-kappa B and the mechanism mediated by curcumin,providing a new method for application of curcumin in clinical and theoretical basis.Materials and methodsChoosed rat BV2 microglial cell line cell line and neurons SH-SY5Y cells as the research object,choosed Suitable concentration of RO 10nmol/L,used 10 nmol/L RO processing BV2 cells and SH-SY5Y cells,different concentrations of Cur?1,5,10?mol/L?processing BV2 cells and SH-SY5Y cells,different concentrations of Cur?1,5,10?mol/L?and RO processed BV2 cells and SH-SY5Y cells together,collecting supernatant of different concentrations of Cur?1,5,10?mol/L?and RO processed BV2 cells groups as conditioned medium add to untreated SH-SY5Y cells and set up the blank control group.Using tetramethyl azo azole salt method detected two kinds of cell vitality,Western blot detected BV2 cells within the RAGE,nf-kappa B expression,ELISA method detected the contents of BV2 cell culture supernatant on inflammatory factor,MTT method to detect conditioned medium of SH-SY5Y cells proliferation activity.The results1.MTT detected the cell viability:regardless of 10nmol/L RO exist,Cur different concentrations?0 to 10?mol/L?after the intervention did not affect the BV2cell and SH-SY5Y cell vitality,no significant differences between groups.2.Western blot detected RAGE and nf-kappa B protein expression:group compared with the blank control group,RAGE and nf-kappa B protein expression of RO group increased significantly?P<0.01?,compared with RO group,the amount of protein expression of RAGE and nf-kappa B of Cur treatment groups significantly reduced?P<0.05?.3.ELISA method detected BV2 cell culture supernatant inflammatory factor levels:compared with the blank control group,the production of inflammatory markers of RO group increased significantly?P<0.01?,compared with RO group,inflammatory factors of production of Cur treatment groups gradually reduced with the increase of concentration?P<0.05?.4.MTT method detected conditioned medium of SH-SY5Y cell proliferation activity:the cell viability of SH-SY5Y cells which were handled by conditioned medium,compared with control group,the RO group decreased significantly?P<0.01?,but the cell vitality of Cur processing groups increaseed compared with RO group,statistically significant difference?P<0.05?.Conclusion1.10 nmol/L RO for BV2 cells and SH-SY5Y cells had no obvious neurotoxicity,the cell viality of BV2 cells and SH-SY5Y cells had no obvious effect when the concentrations of Cur changed from the 0 to10?mol/L.2.RO can obviously increase RAGE and nf-kappa B protein expression of BV2 cells,Cur breaked the RAGE and nf-kappa B protein expression,with the increase of concentration the weakening effect increased.3.RO can induce BV2 cells to secrete inflammatory factor,Cur can suppress the secretion of inflammatory cytokines of BV2 cells,with the increase of concentration this inhibitory ability enhanced.4.Cur could inhibit RAGE and nf-kappaB protein expression,thereby inhibited RAGE/the nf-kappaB pathway and downstream inflammation,thus played a part in the PD neural protection.