Eucommia Bark Alcohol Extract Induced Rat Bone Marrow Mesenchymal Stem Cells Into Bone Differentiation Of Wnt Signaling Pathways In The Process Of Research

Objective:Observation of eucommia bark alcohol extract induced rat bone marrow mesenchymal stem cells into bone differentiation pathways related to Wnt signal pathway in gene expression changes.Methods:Preparation of eucommia bark into salt,eucommia bark,take good preparation of eucommia bark powder,50grams of salt by60%ethanol extraction and column chromatography and other methods,eucommia bark alcohol extract.Will be2months SD rats after cervical vertebra dislocation method executed in volume fraction of70%ethanol solution of5-10minutes,and under aseptic conditions will rat femur and tibia bone marrow mesenchymal stem cells in the isolation,culture and extend the use of third-generation SD rat BMSCs,the inoculation to the culture plate6holes,each hole1×103cells,set up induced group and the negative control group,normal culture medium (including the volume fraction of7.5%fetal bovine serum DMEM/F12) training for24h,24h after induction group into its culture medium (including the volume fraction of7.5%fetal bovine serum DMEM/F12(1:1) plus1/1000of eucommia bark alcohol extract concentration),and the negative control group continue to use the normal culture medium.The RT-qPCR method to induce4h,8h,24h and72h when Wnt signaling pathway related genes (Fzd and LRP series, beta within Wnt catenin, nuclear receptor regulation target genes series and Wnt inhibitors,etc.) expression were determined.Results:Induction groups compared with negative control group,its group after4h, beta-catenin expression increases18.65times, back to normal after8h, significantly reduced after24h,72h expression increased7.61times; Higher induction group after8h Fzd3express,and continue to72h,TCF3expression reduced significantly and continued to72h.Conclusions:Eucommia ulmoides results suggest Wnt signal pathway may be involved in the osteogenetic differentiation of bone marrow mesenchymal stem cells.Meaning:By retrieving literatures,promote the differentiation of MSCs into bone cortex eucommiae Wnt signal transduction pathways is unconfirmed.Through experimental research,eucommia bark in the process of promoting the osteogenetic differentiation of bone marrow mesenchymal stem cells Wnt signaling pathways related gene expression changes obviously,to induce the differentiation of MSCs into bone molecular mechanism and signal pathway of accumulation of traditional Chinese medicine experiment data.