Preliminary Exploration The Role Of Wnt/β-catenin Pathway In Proliferation Of Neural Stem Cells Induced By High-intensity Pulsed Electromagnetic Fields

Object：Comparing β-catenin levels of neural stem cells which were inhibited by β-catenin shRNA1and β-catenin shRNA2lentivirus.Methods:1.Obtain neural stem cells（NSCs）：derived from subventricular zone（SVZ） of newborn SD rats，and cultured for14days using serum-free medium.2.Building β-catenin shRNA1, β-catenin shRNA2and negative control lentivirus.3.Preparation for the transfection: Grouped the neural stem cells into β-cateninshRNA1, β-catenin shRNA2and negative control group, calculated the transfectefficiency of each group.4. When MOI=5, using Real Time PCR and Western Blotto test the levels of β-catenin in NSCs of the three groups.Results:1.When transfection in advance of low MOI, the β-catenin shRNA1andnegative control transfection efficiency were83.89±10.8%,72.92±20.8%,78.48±18.1%, and the transfection efficiency was no significant difference (p>0.05).2.When MOI=5, the β-catenin mRNA levels inhibited by the β-catenin shRNA1decreased63.56%compared with the blank group. While theβ-catenin shRNA2decreased38.63%compared with the blank group.3. When MOI=5, the β-cateninshRNA1, β-catenin shRNA2inhibited the β-catenin protein, and the β-cateninshRNA1had the strongest inhibitory effect on the expression of β-catenin protein.Conclusion: When MOI=5, the β-catenin shRNA1had the strongest inhibitoryeffect of β-catenin. Object：Comparing theβ-catenin shRNA1lentivirus suppressed β-catenin level ondifferent concentrations.Methods: Obtain neural stem cells（NSCs）：they were derived from subventricularzone （SVZ）of newborn SD rats，and then cultured for14days using serum-freemedium.2. Using Real Time PCR and Western Blot to test the level of β-catenin inneural stem cells when MOI=0,0.1,1,10.3. Ultimately determine the besttransfection concentration of suppressing the β-catenin levels in neural stem cells.Results:1.Compared with MOI=0, when MOI=0.1，1，10, the β-catenin mRNAlevels inhibited by the β-catenin shRNA1decreased to some extent. When MOI=10,the β-catenin shRNA1decreased significantly71.21%compared with the controlgroup.2. When MOI=0.1，1，10, the β-catenin mRNA levels inhibited by theβ-catenin shRNA1decreased to some extent, especially when MOI=10.Conclusion:1. When transfected by β-catenin shRNA1lentivirus, the inhibition ofβ-catenin were significantly lower in MOI=10than other concentration.2. TheMOI=10was the optimal transfection concertration, and we can obtain the neuralstem cells of which β-catenin expression was inhibited by β-catenin shRNA1lentivirus for follow-up study. Object： Using the neural stem cells of which β-catenin expression was inhibited,then preliminary explore the role of Wnt/β-catenin pathway in proliferation of neuralstem cells induced by the High-intensity Pulsed Electromagnetic fields（HIPEMF）.Methods:1. Obtain neural stem cells (NSCs): they were derived fromsubventricular zone (SVZ) of newborn SD rats, and then cultured for14days usingserum-free medium.2. Using the β-catenin shRNA1lentivirus to transfect the neuralstem cells on the concertration of MOI=10.3. The NSCs were grouped into thenegative control group and experimental group, after that they were exposed in theHIPEMF with stimulus condition of4T,0.1Hz,8times.4. After that these cellswere detected with the CCK8assay to estimate the proliferation of neural stem cells.Results: The first day and the seventh day after Neural stem cells were exposed inthe HIPEMF, the proliferation of the negative control NSCs group were higher thanthe experimental group (p <0.05).Conclusion: After using RNAi method to inhibit β-catenin expression in neural stemcells, it did not present the proliferation of neural stem cells after exposed inHIPEMF. It took a hint that the Wnt/β-catenin pathway play a role in proliferation ofneural stem cells induced by High-intensity Pulsed Electromagnetic fields.