摘要
目的探讨腺相关病毒介导的klotho(KL)基因表达对去势骨质疏松大鼠的调控作用。方法 SD雌性大鼠随机分为假手术组(S组)和手术组,外科去势术后12周再随机分为模型组(O组)、17β-雌二醇组(E组)、KL基因组(KO组)和空质粒组(GO组),实验12周后处死。取股骨、胫骨测骨密度;冰冻切片及免疫组化法观察肾KL荧光及KL蛋白表达;RT-PCR和免疫组化法检测骨Runx2、MMP-13 mRNA及蛋白表达;HE染色观察骨组织形态学变化。结果 KO组和E组骨密度高于O组和GO组(P<0.05);KO组大鼠肾有小鼠KL基因特异性表达;与O组相比,KO组Runx2 mRNA表达明显上调,MMP-13 mRNA表达显著下调(P<0.05);免疫组化分析KO组Runx2吸光度值为411±96,显著高于O组的353±50(P<0.05);KO组MMP-13吸光度值为397±84,显著低于O组的656±89(P<0.05)。KO组、E组和S组大鼠骨小梁排列紧密,连接成网,形态结构较完整,明显优于O组和GO组。结论 KL基因表达上调可减缓去势大鼠骨质疏松症的发展及骨组织微结构的破坏,提示KL基因可能在骨质疏松症的发展中扮演重要角色。
Objective To research the effect of the recombinant adeno-associated virus vector containing klotho gene delivery on the regulating of osteoporosis in ovariectomized rats.Methods Female SD rats were randomly divided into sham operation group（S group） and model group.Model was successfully constructed with ovariectomy after 12 weeks,they were randomly divided into model group（O group）,17β-estradiol（E group）,klotho gene group（KO group）,empty vector group（GO group）,all were sacrificed after 12 weeks.Bone mineral density（BMD） of the femurs and tibia were measured.The fluorescent expression of renal klotho was observed by Cryo-sectioning technique.The Runx2 and MMP-13 mRNA expression of bone tissue were detected by reverse transcription-polymerase chain reaction（RT-PCR）.Expression of klotho protein in kidney and Runx2,MMP-13 protein in bone were detected by immunohistochemistry.Bone morphological changes of the different groups were observed byHE staining.Results BMD of KO and E groups were significantly higher than those in the O and GO groups.Specific expression of mouse klotho was seen only in KO group.Renal klotho protein in KO group increased.Compared to O group,the expression of Runx2 mRNA increased greatly,but the expression of MMP-13 mRNA decreased in bone tissue of KO group.The average absorbance of Runx2 was 411±96 in KO group.The average absorbance of MMP-13 was 397±84 in KO group,the value of O group was 656±89,so KO group showed a significantly lower than those in O group.The trabecular bones in KO,E,S groups tightly packed and interconnected to form a network.Conclusions Klotho gene delivery attenuates the progression of osteoporosis and bone microstructure damage in ovariectomized rats,suggesting that klotho gene may play a role in the progression of osteoporosis.
出处
《基础医学与临床》
CSCD
北大核心
2012年第5期487-492,共6页
Basic and Clinical Medicine
基金
国家自然基金(30672212)
重庆市卫生局医学科研计划项目(2010-2-079)
