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。
AIM:To investigate the effect of the overexpression of voltage-gated chloride channel family protein3( 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-3overexpressed( 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 detected.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.