CKIP-1对模拟失重诱导的成骨细胞微丝变化的影响  

The Effect of CKIP-1 on Microfilament Changes in Osteoblasts Induced by Simulated Weightlessness

作　　者：王脉桃 宋卫红[1] 周华 万倩 惠辉[1] WANG Maitao;SONG Weihong;ZHOU Hua(The First People's Hospital of Chenzhou City,Chenzhou Hunan 423000)

机构地区：[1]郴州市第一人民医院,湖南郴州423000

出　　处：《医学临床研究》2024年第4期500-503,共4页Journal of Clinical Research

基　　金：郴州市科学技术局科技发展计划项目(ZDYF2020018)。

摘　　要：【目的】探讨酪蛋白激酶结合蛋白1(CKIP-1)对模拟失重诱导的成骨细胞微丝变化的影响。【方法】选择小鼠成骨样细胞株(MC3T3-E1)进行研究,借助回转器模拟微重力环境,通过慢病毒感染构建CKIP-1过表达模型,将细胞分为a组(微重力组)、b组(空载慢病毒及微重力组)、c组(CKIP-1过表达慢病毒及微重力组)、d组(CKIP-1过表达慢病毒组)。使用FITC标记的鬼笔环肽对细胞进行荧光染色,在激光共聚焦显微镜下观察细胞微丝的变化,同时通过qRT-PCR技术测定CKIP-1 mRNA、ALP mRNA及RUNX2 mRNA的表达情况。【结果】每组细胞的微丝结构均呈现不同程度的解聚,而且张力纤维可见紊乱排列,并数量下降,以CKIP-1过表达慢病毒感染的成骨细胞模拟微重力组改变最为明显。c组、d组CKIP-1 mRNA表达水平高于a组、b组,差异有统计学意义(P<0.05)。c组成骨细胞RUNX2 mRNA、ALPL mRNA表达水平均低于a组、b组、d组,差异有统计学意义(P<0.05)。a组、b组、d组成骨细胞RUNX2 mRNA、ALPL mRNA表达水平比较,差异无统计学意义(P>0.05)。【结论】CKIP-1过表达可能会进一步加重微重力条件下成骨细胞骨架微丝的解聚,进而抑制成骨细胞增殖及分化。【Objective】To investigate the effect of CKIP-1 on microfilament changes induced by simulated weightlessness in osteoblasts.【Methods】We selected mouse osteoblast like cell line(MC3T3-E1)for the study,simulated microgravity 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 overexpression 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 under laser confocal microscopy.The expression of CKIP-1 mRNA,ALP mRNA,and RUNX2 mRNA was also measured using qRT-PCR technology.【Results】The microfilament structure of each group of cells showed varying degrees of depolymerization,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 difference 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 ALPL 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 conditions,thereby inhibiting the proliferation and differentiation of osteoblasts.

关 键 词：成骨细胞/超微结构 失重模拟 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]