ClC-3基因过表达对小鼠骨量和骨结构的影响

摘要

[ ABSTRACT] AIM:To investigate the effect of the overexpression of voltage-gated chloride channel family protein 3 ( ClC-3) gene on bones of mice .METHODS: The tail gene detection assay was used to confirm the overexpression of ClC-3.The male FVB mice of three months old were divided into two groups , the wild type ( WT) group and the ClC-3 overexpressed (ClC-3 transgene) group.The body weight, length and weight of the right tibias were measured .The upper and middle parts of the tibias were dissected , decalcified, paraffin-imbed, sectioned and stained with HE staining .The bone morphology metrology was used to analyze the changes of bone structures .The percent trabecular area (%Tb.Ar), trabecular number ( Tb.N) , trabecular width ( Tb.Wi) and trabecular separation ( Tb.Sp) of cancellous bone in the upper part of the tibia were measured.The total tissue area (T.Ar), cortical area (Ct.Ar), percent cortical area (%Ct.Ar), marrow area ( Ma.Ar) and percent marrow area (%Ma.Ar) of the cortical bone in the middle part of the tibia were detec-ted .RESULTS:The wild type mice and the ClC-3-overexpressed mice were verified by the tail gene detection assay . Compared with WT group , the body weight and the length and weight of the tibia were decreased in ClC -3 transgene mice (P<0.05).In the cancellous bones of ClC-3 transgene mice, the%Tb.Ar and Tb.Wi were decreased (P<0.05), the Tb.Sp was increased (P<0.05) and the Tb.N was not significantly changed .In the cortical bones of ClC-3 transgene mice, the T.Ar, Ct.Ar and%Ct.Ar were decreased (P<0.05), the%Ma.Ar was increased (P<0.05), and the Ma. Ar was not significantly changed .CONCLUSION:ClC-3 overexpression may lead to the reduction of the bone mass and the destructure of the cancellous and cortical bones .The results suggest that ClC-3 may be involved in the regulation of bone resorption and/or formation.%目的：研究过表达电压门控氯通道家族蛋白成员3（voltage-gated chloride channel family protein 3， ClC-3）基因对小鼠骨骼的影响。方法：3月龄雄性FVB小鼠用鼠尾基因检测法确定基因型后分为2组：野生型（WT）组和ClC-3过表达（ClC-3 transgene）组，每组各8只。分别测量2组小鼠体重，右侧胫骨长度和重量。取胫骨上段和中段进行脱钙，石蜡包埋，切片，HE染色后，采用骨形态计量学分析胫骨上段松质骨的骨小梁面积百分数（percent trabecular area，％Tb．Ar）、骨小梁数量（trabecular number， Tb．N）、骨小梁宽度（trabecular width， Tb．Wi）、骨小梁分离度（trabecular separation， Tb．Sp）和胫骨中段皮质骨的骨组织总面积（total tissue area， T．Ar）、皮质骨面积（cortical area， Ct．Ar）、皮质骨面积百分数（percent cortical area，％Ct．Ar）、骨髓腔面积（marrow area， Ma．Ar）、骨髓腔面积百分数（ percent marrow area ，％Ma．Ar）。结果：小鼠经鼠尾基因检测后确认为野生型或ClC-3过表达型。与WT组相比，ClC-3 transgene组小鼠体重及胫骨长度重量均减小（ P＜0.05）；松质骨的％Tb．Ar和Tb．Wi均减小（P＜0.05），Tb．Sp增大（P＜0.05），Tb．N的变化无统计学意义；皮质骨的T．Ar、Ct．Ar和％Ct．Ar均减小（P＜0.05），％Ma．Ar增大（P＜0.05），Ma．Ar的变化无统计学意义。结论：ClC-3过表达导致小鼠的皮质骨和松质骨骨量减少，骨结构变差，提示ClC-3可能参与了骨形成和／或骨吸收。