The Effect of CKIP-1 on Microfilament Changes in Osteoblasts Induced by Simulated Weightlessness
[Objective]To investigate the effect of CKIP-1 on microfilament changes induced by simulated weightless-ness in osteoblasts.[Methods]We selected mouse osteoblast like cell line(MC3T3-E1)for the study,simulated microgravi-ty environment with a gyrator,and constructed a CKIP-1 overexpression model through lentivirus infection.The cells were divided into group a(microgravity group),group b(empty lentivirus and microgravity group),group c(CKIP-1 overex-pression lentivirus and microgravity group),and group d(CKIP-1 overexpression lentivirus group).Fluorescence staining was performed on cells using FITC labeled ghost pen cyclic peptides,and changes in cell microfilaments were observed un-der laser confocal microscopy.The expression of CKIP-1 mRNA,ALP mRNA,and RUNX2 mRNA was also measured u-sing qRT-PCR technology.[Results]The microfilament structure of each group of cells showed varying degrees of depoly-merization,and the tension fibers were disorderly arranged and the number decreased.The most significant changes were observed in the simulated microgravity group of osteoblasts infected with CKIP-1 overexpression lentivirus.The expression level of CKIP-1 mRNA in the group c and the group d was higher than that in the group a and the group b,and the differ-ence was statistically significant(P＜0.05).The expression levels of RUNX2 mRNA and ALPL mRNA in bone cells composed of the group c were lower than those in the group a,the group b,and the group d,with statistical significance(P＜0.05).There was no statistically significant difference in the expression levels of RUNX2 mRNA and AL-PL mRNA in bone cells between the group a,the group b,and the group d(P＞0.05).[Conclusion]Overexpression of CKIP-1 may further exacerbate the depolymerization of cytoskeletal microfilaments in osteoblasts under microgravity condi-tions,thereby inhibiting the proliferation and differentiation of osteoblasts.
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