Preliminary Study On The Repair Mechanism Of Piezo1 In Femoral Head Necrosis Of Kidney Deficiency And Blood Stasis Type

ObjectiveTo study the effect of Piezo1,a mechanical sensitivity channel,on osteoblast apoptosis and angiogenesis in steroid-induced osteonecrosis of the femoral head(ONFH)rats and explore the mechanism,so as to provide a new idea for the repair mechanism of femoral head necrosis.MethodsFifty-six male adult Wistar rats(12 weeks old)were divided into 4 groups(n=16),two rats in each group were randomly selected as the basis for the success of pre experiment and modeling,including steroid hormones(steroid,STE)combined with Piezo1 silencing carrier(si RNA-Piezo1,siP1)(STE+siP1 group),STE combined with silence negative control group(STE+si NC group),STE group and control group.Collect femoral skull tissue.TUNEL method detects apoptosis rate,three-dimensional microangiography observes the changes in the amount of blood vessels in the femoral head,q PCR detects the m RNA expression of Piezo1,apoptosis markers Bax,Caspase-3,and Bcl-2,Western blotting was used to detect the expression of Piezo1,angiogenesis markers VEGFR2,CD3 and Yap1/β-catenin signaling protein.Culture human osteoblast cell line(h FOB 1.19)and umbilical vein endothelial cell line(HUVEC),the cells were treated with recombinant human YAP1 protein(Rh-YAP1)and β-catenin activator(ICG-001).TUNEL method was used to detect the apoptosis rate of h FOB 1.19.Tube structure formation experiment was used to evaluate the angiogenesis ability of HUVEC.ResultssiP1 successfully inhibits the expression of Piezo1 in the femoral head tissue in the ONFH rat model,and siP1 inhibits the apoptotic rate of femoral head tissue and the expression of apoptosis markers,partially restores the loss of femoral head blood vessels in the ONFH model,and up-regulates angiogenesis Marker level.siP1 inhibits the expression of Yap1 and β-catenin protein.Using Rh-YAP1 and ICG-001 to activate the Yap1/β-catenin signaling pathway resulted in a significant increase in the apoptosis of h FOB 1.19,but the angiogenesis ability of HUVEC cells was decreased.ConclusionIn the ONFH model,silencing Piezo1 can improve osteoblast apoptosis and promote angiogenesis by inhibiting the Yap1/β-catenin signaling pathway,it can provide new discoveries for the molecular biological mechanism of femoral head necrosis repair.