Icariin, Curcumin And Piperine Prevent The Progress Of Early AD Pathology

Objective: To explore the protective effects of a Chinese medicine compound formula(consisted of icariin, curcumin and piperine, ICP) repair the energy metabolism inhibitory of 3x Tg AD mouse 、 primary neuronal celland mouse \neuroblastoma N2 a cells,we choose mitochondria as the early AD target spot,try to expound the mitochondrial Energetics and dynamics in treating AD molecular mechanism,and giving the preparation to exploit Chinese patent medicine to prevent AD.Method: 1、in exo experiment:1)、The preparation is establishing primary cortical neuronal cell cultures from non-transgenic mouse(NTg、Normal group)、 3x Tg-AD mouse(Tg、control group) and Tg neurons were treated with ICP formula(Tg+ICP、experimental group)for 24 h. Using immunocytochemistry 、The laser confocal observation、Protein immunoblot technique and ATP evel detection technology,etc to analysis and comparison the changes between Normal group 、control group and experimental group in AD pathology、Mitochondrial energy metabolism and dynamic balance and synapse-associated proteins,etc;2)、The preparation is establishing the AD model cells--mouse neuroblastoma N2 a cells( Normal group) and N2asw(control group),whith N2 a cells were stably transfected with human “ Swedish” mutated form of APP695, and the N2 asw cells were treated with ICP formula(N2asw+ICP、experimental group)for 24 h. Using immunocytochemistry 、The laser confocal observation 、 Protein immunoblot technique and ATP evel detection technology,etc to analysis and comparison the changes between Normal group 、control group and experimental group in AD pathology、Mitochondrial energy metabolism and dynamic balance and synapse-associated proteins,etc。Also,using co-ip technology to preliminary discuss the interaction relationshipbetween mitochondrial protein and phosphorylated tau protein.2、in vivo experiment:We prepare fodder that contain ICP,and feed AD model mice with the fodder from 2 month-age to 8 month-age(Tg+ICP、experimental group)comparing the non-transgenic mouse(Ng、Normal group) and AD model mice(Tg、control group),feeding none ICP fodder 。 Then using he Morris water maze experiment、western blotting to compare the differents between Normal group, control group and experimental group about AD pathology 、 Mitochondrial energy metabolism and dynamic balance and synapse-associated proteins,etc。Results : 1、in exo experiment:From the mouse primary neuronal cell to AD model cell,it is the Co-verification that Chinese compound ICP has certain effection in preventing and interfering the early AD pathological process and mitochondrial injury。Using Western bloting and ROS detection kit have found that ICP could weaken Aβand secretions generate and could inhibit tau protein hyperphosphorylation,reduce ROS produce.The changes about mitochondria like:ICP increase ATP concentration and release lactic dehydrogenase and recover the balance of the mitochondrial energy metabolism。Also,Western bloting have showed that ICP restore mitochondria dynamic equilibrium。Examine synapse by Western blot and cell immunofluorescence location, found that ICP could increase the expression of postsynaptic density protein(PSD95),protect synapse。Further,Using western blot to state the time-dependent of ICP.。2、in vivo experiment:Morris water maze have showed that ICP could affect early AD mouse spatial learning and memory ability。Respectively,Using western blot test hippocampus and cortex tissue of mouse found that ICP may reduce Aβgeneration and deposition by lower BACE-1 concentration level。In the meantime,reduce tau protein hyperphosphorylation,ensure normalization of microtubule-associated protein。ICP recover the balance of the mitochondrial energy metabolism and increase the expression of postsynaptic density protein(PSD95),inprove mice cognition and memory,protect neuron.Conclusion:The Chinese medicine formula ICP may intervene Aβ abnormal accumulation and interaction between tau protein to reduce phosphorylation and recover synapse energy supply by repair mitochondrial energy metabolism injury anddynamics out-of-balance。So,The Chinese medicine formula ICP may have some therapeutic effect to prevent early AD。...