慢性肾功能不全幼鼠胫骨生长板软骨细胞初级纤毛表达对Wnt/β 信号通路的影响  

Effect of primary cilia expression rate on Wnt/β signaling pathway in tibial growth platechondrocytes from chronic renal insufficiency young rats

作　　者：王小健[1] 张健 刘芳[2] 周晓霜[3] 赵中涛[1] 郭秀生[1] 宋洁富[1] 苏云星[1] 李荣山[3] Wang Xiaojian;Zhang Jian;Liu Fang;Zhou Xiaoshuang;Zhao Zhongtao;Guo Xiusheng;Song Jiefu;Su Yunxing;Li Rongshan(Department of Orthopaedic Surgery,Shanxi Provincial People's Hospital,Taiyuan030012,China;MedicalInformation and Data Center,Shanxi Provincial People Hospital,Taiyuan030012,China;Department of Nephrology,Shanxi Provincial People’s Hospital,Taiyuan030012,China)

机构地区：[1]山西省人民医院骨科,太原030012 [2]山西省人民医院医学情报与图书资料室,太原030012 [3]山西省人民医院肾内科,太原030012

出　　处：《中华医学杂志》2019年第46期3645-3651,共7页National Medical Journal of China

基　　金：山西省自然科学基金(201701D121161);山西省重点研发计划项目(201803D31160)。

摘　　要：目的观察慢性肾功能不全(CRI)幼鼠胫骨生长板软骨细胞初级纤毛表达对W m/p信号通路的影响。方法雄性2周龄SD幼鼠12只,分为假手术组(暴露左侧输尿管,6只)和CRI组(结扎左侧输尿管,6只)。术后2周处死幼鼠,观测胫骨生长板区软骨细胞初级纤毛及Wnt/|3通路关键蛋白P-连环蛋白(p-catenin)的表达。提取两组幼鼠胫骨生长板软骨细胞进行体外培养至P 3代,检测细胞初级纤毛及P-catenin表达。应用W nt/p通路激动剂、拮抗剂干预CRI组软骨细胞,检测细胞初级纤毛表达率;应用无血清培养、水合氯醛干预CRI组软骨细胞,检测p-catenin表达水平。结果与假手术组相比,CRI组胫骨生长板区软骨细胞初级纤毛表达率增高[(17.5±7.7)%比(8.7±3.6)%,(=3.237,P=0.005],(3-catenin表达免疫组化组织学评分增高[(5.1士0.7)分比(1.9±0.8)分,《=6.731,P<0.001]。软骨细胞体外培养在CRI组细胞初级纤毛表达率增高[(20.9±8.1)%比(11.8±4.7)%J=3.073,P=0.007],Western印迹结果显示p-catenin表达增卨(0.49±0.丨2比0.25±0.11,t=3.297,P=0.0U)。应用Wnt/卩通路激动剂和拮抗剂改变P-catenin表达水平后细胞初级纤毛表达率无明显变化[激动剂组(21.3±7.6)%,对照组(20.6±6.8)%,拮抗剂组应用无血清培养、水合氯醛改变细胞初级纤毛表达率后,P-catenin蛋白表达量改变(无血清组0.61±0.23,对照组0.39±0.24,水合氯醛组0.15±0.11,F=6.476,P=0.012),且与细胞初级纤毛表达率呈正相关。结论肾功能不全幼鼠胫骨生长板软骨细胞初级纤毛及Wm/卩通路均呈高表达,初级纤毛可能对W nt/p通路存在正性调控作用。Objective To observe the effect of primary cilia expression rate on Wnt/p signalingpathway in tibial growth plate chondrocytes from chronic renal insufficiency (CRI) young rats. MethodsMale 2-week-old SD rats were randomly divided into two groups: (1) Sham group: only the left ureter wasexposed (n=6);(2) CRI group: the left ureter was ligated (/i=6). Rats were sacrificed 2 weeks after theoperation and the primary cilia expression rate of growth plate chondrocytes and key protein p-catenin inWnt/p signaling pathway were observed in histological section of tibia specimen. Chondrocytes isolated fromgrowth plate in two groups were cultured in vitro to P3 generation. The primary cilia expression rate and thelevel of P-catenin were measured. The primary cilia expression rate was detected by agonists and antagonistsWnt/p signaling pathway in chondrocytes of CRI group. The level of P-catenin was detected by usingserum-free culture and chloral hydrate to intervene chondrocytes in CRI group. Results The primary cilia expression rate of growth plate chondrocytes in histological section of tibia specimen in CRI group washigher than that in Sham group [(17.5±7.7)% vs (8.7±3.6)%, i=3.237, P=0.005], and the level of P-cateninwas higher in CRI group (5.1 ±0.7 vs 1.9±0.8, f=6.731, P<0.001). The primary cilia expression rate of growthplate chondrocytes cultured h t^ro in CRI group was higher than that in Sham group [(20.9±8.1)% vs (11.8士4.7)%, f=3.073, P=0.007], and the level of p-catenin was higher in CRI group (0.49±0.12 vs 0.25±0.11,艺=3.297, P=0.011). There was no significant change in primary cilia expression rate after intervention by usingWnt/p signaling agonists and antagonists to change the level of P-catenin [agonists group: (21.3 ±7.6) % ,control: (20.6 ±6.8)% , antagonists group: (22.4 ±6.2)% ,厂=0.173,P=0.842]. The level of P_catenin wassignificantly changed after intervention by using serum-free culture, chloral hydrate to alter the primary ciliaexpression rate (serum-free culture group: 0.61 ±0.23, co

关 键 词：肾机能不全 生长板 初级纤毛 Wnt/β信号通路 

分 类 号：R73[医药卫生—肿瘤]