摘要
目的探讨慢性肾衰竭(chronic renal failure,CRF)幼鼠胫骨生长板软骨细胞Wnt/β-catenin信号通路对生长板发育的影响。方法雄性4周龄SD幼鼠40只,被随机分为对照组(蒸馏水灌胃,n=20)和CRF组(腺嘌呤150 mg·kg^(-1)·d^(-1)灌胃,n=20),连续灌胃6周后处死全部幼鼠,比较两组幼鼠胫骨长度。micro-CT扫描仪测量胫骨近端生长板宽度,组织切片番红固绿染色检测胫骨生长板宽度。提取幼鼠胫骨生长板软骨细胞进行体外培养至第3代,免疫组织化学染色法检测软骨细胞Ⅱ型胶原(CollagenⅡ)、基质金属蛋白酶13(matrix metalloproteinase-13,MMP-13)及β联蛋白(β-catenin)的表达。Western印迹法检测软骨细胞CollagenⅡ、MMP-13、β-catenin蛋白表达水平。结果与对照组相比,CRF组幼鼠胫骨长度较短[(27.32±5.81)mm比(35.43±3.61)mm,t=5.226,P<0.001],生长板宽度较窄[(0.72±0.22)mm比(1.13±0.27)mm,t=5.096,P<0.001]。软骨组织番红固绿染色结果显示,CRF组幼鼠生长板相对宽度较对照组窄(t=6.744,P<0.001)。免疫组化及Western印迹结果示,与对照组比较,CRF组软骨细胞CollagenⅡ蛋白表达减少(t=8.212,P<0.001),MMP-13(t=13.091,P<0.001)和β-catenin(t=7.534,P<0.001)蛋白表达增多。结论CRF幼鼠胫骨生长板软骨细胞Wnt/β-catenin通路相关蛋白呈高表达状态,是软骨细胞加速退变分化、生长板出现闭合趋势的原因之一。
Objective To explore the effect of Wnt/β-catenin signaling pathway on growth plate development of tibial growth plate in young chronic renal failure(CRF)rats.Methods Four-week-old male SD rats were randomly divided into control group and CRF group(n=20/per group).Control group was intragastric administration with distilled water,and CRF group was given adenine suspension(150 mg·kg^(-1)·d^(-1)).All the young rats were sacrificed after continuous gavages for 6 weeks.The full length of tibia was compared between the two groups.The width of tibia proximal growth plates was measured by micro-CT scanning,and the width of the growth plate was also measured in histological sections.Chondrocytes isolated from growth plate in two groups were cultured in vitro to P3 generation.Immunohistochemical staining was used to detect the expression of collagenⅡ,matrix metalloproteinase 13(MMP-13)andβ-catenin in chondrocytes.Western blotting was used to detect the protein expressions of collagenⅡ,MMP-13 andβ-catenin.Results Compared with the control group,the tibial length of rats in the CRF group was shorter[(27.32±5.81)mm vs(35.43±3.61)mm,t=5.226,P<0.001],the width of growth plate in micro-CT picture was more narrow[(0.72±0.22)mm vs(1.13±0.27)mm,t=5.096,P<0.001],and the relative width of the growth plate was also more narrow(t=6.744,P<0.001)in histological sections.The results of immunohistochemistry and Western blotting showed the expressions of collagenⅡin the CRF group decreased significantly(t=8.212,P<0.001),MMP-13(t=13.091,P<0.001)andβ-catenin(t=7.534,P<0.001)increased significantly compared the control group in chondrocytes.Conclusion The Wnt/β-catenin signaling pathway is highly expressed in the tibial growth plate of young rats with chronic renal failure,which leads to accelerated degeneration and differentiation of chondrocytes and a closure tendency of growth plate.
作者
王小健
关贵平
路晓
李亚峰
高艳芳
郑刚
毕宏
苏云星
李荣山
Wang Xiaojian;Guan Guiping;Lu Xiao;Li Yafeng;Gao Yanfang;Zheng Gang;Bi Hong;Su Yunxing;Li Rongshan(Department of Orthopedic Surgery,Shanxi Provincial People's Hospital,Taiyuan 030012,China;Department of Nephrology,Shanxi Provincial People's Hospital,Taiyuan 030012,China;Department of Pathology,Shanxi Provincial People's Hospital,Taiyuan 030012,China)
出处
《中华肾脏病杂志》
CAS
CSCD
北大核心
2021年第2期137-142,共6页
Chinese Journal of Nephrology
基金
山西省自然科学基金(201701D121161)
山西省重点研发计划项目(201803D31160)
中国博士后科学基金(2020M680910)。
