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Objective To investigate the effect of different fluid shear stress (FSS) on the regulation of planar cell polarity (PCP) signaling, and further to explore the relationship among FSS, PCP signaling pathway and ciliogenesis. Methods The hydrodynamic cell model of adjustable FSS was established. qPCR and immunofluorescence were used to detect the mRNA expression of PCP signaling pathway core protein Dvl2 and cilia assembly protein IFT88, cell targeting and co-localization under different FSS. Western blot (WB) was used to detect the protein expression of Dvl2 at 18 h under different FSS. Results The qPCR result showed that compared with 1.5 Pa FSS, under 0.1 Pa FSS, the mRNA expression of Dvl2 was higher at 6 h and 18 h (P＜0.05), significantly higher at 12 h (P＜0.01); the mRNA expression of IFT88 was significantly higher at 18 h (P＜0.01). The WB result showed that compared with 0 h, under 0.1 Pa FSS, the protein expression of Dvl2 was higher at 18 h (P＜0.05), significantly lower under 1.5 Pa FSS (P＜0.01); compared with 1.5 Pa FSS, the protein expression of Dvl2 was higher at 18 h under 0.1 Pa FSS (P＜0.05). The immunofluorescence result showed that the positive expression of Dvl2 increased with the loading time on FSS increasing, and gradually aggregated at a point around the nucleus; the positive expression of IFT88 was gradually transferred from the nucleus to the cytoplasm and aggregated at a point under 0.1 Pa FSS, and gradually decreased and depolymerized under 1.5 Pa FSS. Protein Dvl2 and IFT88 were located in the same position in cells under 0.1 Pa FSS and before 18 h under 1.5 Pa FSS, and colocalization of proteins Dvl2 and IFT88 was not observed after 18 h under 1.5 Pa FSS due to IFT88 depolymerization. Conclusions Laminar FSS played an inhibition on the transduction of PCP signaling pathway and a hindrance on the process of ciliogenesis, while low FSS played a promotion on the transduction. PCP signaling pathway might regulate FSS-induced ciliogenesis by Dvl2.
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Objective To investigate the effects of rabbit carotid artery cannula on atherosclerosis formation and β-catenin expression.Methods Fifteen 2-month-old New Zealand rabbits weighing(2.0±0.2)kg were randomly divided into three groups,high-fat diet group,left common carotid artery cannula group and left common carotid artery cannula plus high-fat diet group,5 cases in each group,and taking the right blood vessels in the left common carotid artery cannula group served as the control group.The animals were sacrificed after 9-week feeding,and the total rabbit carotid artery in each group was taken;the real-time PCR and immunohistochemistry were used to detect β-catenin mRNA and protein expression targeting in rabbit common carotid artery tissue.Results The real-time PCR results showed that the β-catenin mRNA expression in the left common carotid artery cannula group was higher than that in the control group,high fat diet group and the left common carotid artery cannula + high-fat diet group.The immunohistochemistry results showed that,except for the control group,various groups had the β-catenin protein location in the cytoplasma,moreover which in the left carotid artery cannula group and left common carotid artery cannula +high-fat diet group mainly located in the area of intimal hyperplasia.Conclusion β-catenin is highly expressed in the atherosclerotic vessel wall caused by rabbit carotid artery cannula.