骨桥蛋白基因反义寡核苷酸对被动吸烟大鼠骨组织的影响  

Influence of osteopontin gene antisense oligonucleotides on bone and bones of passive smoking rats

作　　者：雷光华[1] 高曙光[1] 覃芙[2] 曾凯斌[1] 徐文硕[1] 徐迈[1] 李康华[1] 

机构地区：[1]中南大学湘雅医院骨科,湖南长沙410008 [2]杭州市余杭区第一人民医院骨科,浙江杭州311100

出　　处：《中国药理学与毒理学杂志》2009年第5期395-403,共9页Chinese Journal of Pharmacology and Toxicology

基　　金：国家自然科学基金资助项目(30300396)~~

摘　　要：目的探讨骨桥蛋白(OPN)在被动吸烟所致大鼠骨吸收中的作用,了解OPN基因反义寡核苷酸(AS-OPN)对大鼠被动吸烟所致骨质疏松的影响,寻求治疗骨质疏松症的可能有效途径。方法2月龄SD大鼠40只,随机分4组:对照组、被动吸烟组、AS-OPN组和有义OPN(S-OPN)组,每组10只。按密室熏烟法给大鼠被动吸烟,同时AS-OPN及S-OPN组大鼠每3d分别iv给予AS-OPN(10μg.L-1)或S-OPN(10μg·L-1)6μL.g-1,吸烟组和对照组给同等剂量的生理盐水,实验持续4个月,然后进行指标的测定。(1)骨代谢生化指标测定:血Ca、血清骨钙素(BGP)和尿Ca/肌酐(Cr)。(2)骨密度测定:测量L3～L6各腰椎骨密度、双侧股骨和肱骨的整体骨密度及其7个感兴趣区(ROI)的骨密度。(3)骨形态计量学测定:①静态参数包括骨小梁面积百分数、骨小梁厚、骨小梁数和骨小梁分离度;②动态参数包括荧光周长百分率和破骨细胞计数。(4)骨生物力学测定:①L4椎体压缩试验测量指标包括弹性模量、最大载荷、骨最大应变和能量吸收。②右股骨三点弯曲试验测量指标包括最大载荷、弹性载荷、最大挠度和弯曲能量;弯曲弹性模量、最大弯曲应力、弯曲刚性系数和弯曲韧性系数。结果与正常对照组比较,吸烟后大鼠骨密度降低、骨量减少、骨强度降低,破骨细胞数和骨吸收增加。与吸烟对照组比较,给予AS-OPN后大鼠的尿Ca/Cr比值降低(0.08±0.01vs0.11±0.02);L3,L4,L5,L6各腰椎骨密度升高〔27.77±1.38vs25.20±1.94;26.80±1.66vs24.25±1.48;27.55±1.61vs24.20±2.13;26.63±1.17vs(22.58±1.69)mg.cm-2〕,左、右侧股骨骨密度升高〔25.39±1.34vs23.26±1.16,26.28±0.92vs(23.30±1.38)mg.cm-2〕;左、右侧肱骨骨密度及其7个ROI的骨密度升高;骨小梁面积百分数、骨小梁厚度、骨小梁数升高〔6.29±0.67vs(5.13±0.54)%,55.82±2.78vs(49.10±4.36)μm,0.73±0.05vs(0.64±0.07)mm〕;骨小梁分离度、破骨细胞计数和荧光周长百分率降�AIM To investigate effects of osteopontin （OPN） in rat bone resorption caused by passive smoking and to understand the influence of osteopontin gene antisense oligonucleotides （AS-OPN） on rat osteoporosis caused by passive smoking and to search for potential effective ways to treat osteoporosis. METHODS Forty Sprague-Dawley（SD） rats （2-month-old） were randomly divided into 4 groups：control,smoking,antisense oligonucleotides （AS-OPN） and sense oligonucleotides （S-OPN） groups. Apart from control group,other group rats were exposed to passive cigarette smoking daily. The rats in AS-OPN group were given 6 μL·g-1 AS-OPN 10 μg·L-1 every 3 d by intravenous injection. The rats in S-OPN group were given 6 μL·g^-1 S-OPN 10 μg·L-1 every 3 d by intravenous injection. The rats in control group and smoking group were given the same dose normal saline every 3 d by intravenous injection. The time had been lasted for 4 months and then they were proceeded to the following measurements：（1） Bone metabolic biochemical indicators were measured including serum calcium（Ca）,serum bone-gla-protein（BGP） and urine calcium/creatinine（Ca/Cr）. （2） Bone mineral density （BMD） was measured including the BMD of 3rd lumbar,4th lumbar,5th lumbar and 6th lumbar vertebrae （L3-L6） and the BMD of two-sides overall femoral and humeral bones and their 7 regions of interest（ROI）. （3） Bone histomorphometry was measured：① Static parameters were including bone trabecula areas percentage,bone trabecula thickness,bone trabecula number and bone trabecula separation degrees.② Dynamic parameters were including fluorescence circumference percentage and osteoclast number. （4） Bone biomechanics was measured：① The indicators of L4 body of vertebra compression testing including elastic modulus,maximal loading,maximum strain of bone and energy absorbability. ② The indicators of right femur three point bending test including the structural mechanics indicators（maximal loading,elasti

关 键 词：烟草烟污染 骨桥蛋白 寡核苷酸类 反义 骨和骨组织 

分 类 号：R580[医药卫生—内分泌]