氟对大鼠腰椎骨组织形态计量学的影响

Objective To study the effect of fluorine on the bone histomorphometry of humbar in rats.Methods Ninety 2-month-old SPF Sparague-Dawley rats,half male and female,were randomly divided into 9 groups:control[(childhood(CS),adult(AS),long-time(NS)]group and drug group[childhood high-fluoride and low-fluoride group(CHS,CLS),adult high-fluoride and low-fluoride(AHS,ALS),long-term high-fluoride and low-fluoride(CLHS,CLLS)].The control group was administered orally with solution of 0.9%NaCl,while the drug group was given orally with different dose of NaF at the same time. Sections of the fifth lumbar were made which was undecalicified for bone histomorphometric analysis, including the percentage of trabecular bone area (% Tb.Ar),trabecular thickness(Tb.Th), trabecular number(Tb.N), trabecular separation(Th.Sp) ; broken trabecular bone area cells (Oc.N), osteoclast perimeter percentage (% Oc.Pm), the percentage of labeled perimeter (% L.Pm), bone mineral apposition rate(MAR), osteoblast perimeter(Ob.PM), trabecular bone perimeter formation rate (BFR/BS),trabecular bone area formation rate (BFR/BV), the total area of bone formation rate (BFR/TV). Results [1]The percentage of Tb.Ar, Tb.Th, Tb.N,%L.Pm, MAR, BFR/BS, BFR/BV and BFR/TV of CHS group [(50.63 ±7.44)%, (150.26 ± 27.51 )μm, (3.44 ± 0.47)N/mm, (50.63 ± 7.44)%, (0.85 ± 0.03)μm/d, (8.45 ± 2.36)μm/d ×100, (381.16 ± 41.62)%/year, (75.07 ± 4.81)%/year] was higher than that of CS group [(29.71 + 9.32)%,(110.93 ± 28.19)μm, (2.68 ± 0.34)N/mm, (24.00 ± 1.22)%, (0.65 ± 0.03)μm/d, (5.43 ± 0.18)μm/d × 100,(141.32 ± 9.29)%/year, (58.14 ± 2.3)%/year, all P < 0.05)]. The %Tb.Ar, Tb.Th, %L.Pm, MAR, BFR/BS,BFR/BV, BFR/TV and Ob.PM of CLS group [(40.76 ± 6.43)%, (164.25 ± 45.65)μm, (42.02 ± 6.12)%, (0.85 ±0.04)μm/d, (8.95 ± 3.73)μm/d × 100, (378.73 ± 35.39)%/year, (73.52 ± 8.71)%/year, (1.41 ± 0.05)μm] were increased (all P < 0.05). [2]Compared with AS group, the %Tb.Ar,Oc.N, %Oc.Pm, %L.Pm, MAR, BFR/BS,BFR/BV and BFR/TV of AHS group[ (50.62 ± 5.76)%, (0.51 ± 0.05)N/mm, (1.13 ± 0.05)%, (42.3 ± 7.02)%,(1.28 ± 0.09)μm/d, (12.91 ± 1.52)μm/d × 100, (390.12 ± 43.56)%/year, (65.21 ± 22.13)%/year] was higher than that of AS group[ (42.73 ± 5.22)%, (0.41 ± 0.17)N/ram, (0.77 ± 0.52)%, (28.43 ± 6.93)%, (0.80 ± 0.03)μm/d, (9.83 ± 1.44)μm/d × 100, (324.43±53.44)%/year and(48.35 ± 9.36)%/year, all P < 0.05)] . The %Tb.At, Oc.N, %Oc.Pm, %L.Pm, MAR, BFR/BS, BFR/BV and BFR/TV of ALS group [(51.14 ± 6.22)%, (0.49 ±0.61)N/mm, (1.17 ± 0.11)%, (45.06 ± 6.92)%, (1.39 ± 0.08)μm/d, (12.87 ± 1.35)μm/d × 100, (394.6 ±50.23)%/year and(66.31 ± 18.93)%/year] were higher than that of AS group(P < 0.05) .[3] The Ob.PM ,Oc.N and %Oc.Pm of CLHS group[ (1.47 ± 0.27)μm, (0.58 ± 0.13)N/mm, (1.14 ± 0.07)%] were obviously increased(P <0.05), as compared with NS group [ (0.82 ± 1.20)μm, (0.42 ± 0.25)N/mm and (0.75 ± 0.64)%, all P < 0.05].Conclusions The short-term administration of NaF on rats in the growing period increases the bone formation and osteoblast activities of young rats and adult rats. The long-term administration of NaF on rats does not increase the bone formation of rats in growth period. The osteoblast activities as well as the bone absorption of lumbar vertebra were strengthened. The likelihood of bone fracture became larger. The negative effects on bone metabolism and bone quality of rats were gradually displayed along with the prolongation of sodium fluoride usage.     

氟对大鼠腰椎骨组织形态计量学的影响

Objective To study the effect of fluorine on the bone histomorphometry of humbar in rats.Methods Ninety 2-month-old SPF Sparague-Dawley rats,half male and female,were randomly divided into 9 groups:control[(childhood(CS),adult(AS),long-time(NS)]group and drug group[childhood high-fluoride and low-fluoride group(CHS,CLS),adult high-fluoride and low-fluoride(AHS,ALS),long-term high-fluoride and low-fluoride(CLHS,CLLS)].The control group was administered orally with solution of 0.9%NaCl,while the drug group was given orally with different dose of NaF at the same time. Sections of the fifth lumbar were made which was undecalicified for bone histomorphometric analysis, including the percentage of trabecular bone area (% Tb.Ar),trabecular thickness(Tb.Th), trabecular number(Tb.N), trabecular separation(Th.Sp) ; broken trabecular bone area cells (Oc.N), osteoclast perimeter percentage (% Oc.Pm), the percentage of labeled perimeter (% L.Pm), bone mineral apposition rate(MAR), osteoblast perimeter(Ob.PM), trabecular bone perimeter formation rate (BFR/BS),trabecular bone area formation rate (BFR/BV), the total area of bone formation rate (BFR/TV). Results [1]The percentage of Tb.Ar, Tb.Th, Tb.N,%L.Pm, MAR, BFR/BS, BFR/BV and BFR/TV of CHS group [(50.63 ±7.44)%, (150.26 ± 27.51 )μm, (3.44 ± 0.47)N/mm, (50.63 ± 7.44)%, (0.85 ± 0.03)μm/d, (8.45 ± 2.36)μm/d ×100, (381.16 ± 41.62)%/year, (75.07 ± 4.81)%/year] was higher than that of CS group [(29.71 + 9.32)%,(110.93 ± 28.19)μm, (2.68 ± 0.34)N/mm, (24.00 ± 1.22)%, (0.65 ± 0.03)μm/d, (5.43 ± 0.18)μm/d × 100,(141.32 ± 9.29)%/year, (58.14 ± 2.3)%/year, all P < 0.05)]. The %Tb.Ar, Tb.Th, %L.Pm, MAR, BFR/BS,BFR/BV, BFR/TV and Ob.PM of CLS group [(40.76 ± 6.43)%, (164.25 ± 45.65)μm, (42.02 ± 6.12)%, (0.85 ±0.04)μm/d, (8.95 ± 3.73)μm/d × 100, (378.73 ± 35.39)%/year, (73.52 ± 8.71)%/year, (1.41 ± 0.05)μm] were increased (all P < 0.05). [2]Compared with AS group, the %Tb.Ar,Oc.N, %Oc.Pm, %L.Pm, MAR, BFR/BS,BFR/BV and BFR/TV of AHS group[ (50.62 ± 5.76)%, (0.51 ± 0.05)N/mm, (1.13 ± 0.05)%, (42.3 ± 7.02)%,(1.28 ± 0.09)μm/d, (12.91 ± 1.52)μm/d × 100, (390.12 ± 43.56)%/year, (65.21 ± 22.13)%/year] was higher than that of AS group[ (42.73 ± 5.22)%, (0.41 ± 0.17)N/ram, (0.77 ± 0.52)%, (28.43 ± 6.93)%, (0.80 ± 0.03)μm/d, (9.83 ± 1.44)μm/d × 100, (324.43±53.44)%/year and(48.35 ± 9.36)%/year, all P < 0.05)] . The %Tb.At, Oc.N, %Oc.Pm, %L.Pm, MAR, BFR/BS, BFR/BV and BFR/TV of ALS group [(51.14 ± 6.22)%, (0.49 ±0.61)N/mm, (1.17 ± 0.11)%, (45.06 ± 6.92)%, (1.39 ± 0.08)μm/d, (12.87 ± 1.35)μm/d × 100, (394.6 ±50.23)%/year and(66.31 ± 18.93)%/year] were higher than that of AS group(P < 0.05) .[3] The Ob.PM ,Oc.N and %Oc.Pm of CLHS group[ (1.47 ± 0.27)μm, (0.58 ± 0.13)N/mm, (1.14 ± 0.07)%] were obviously increased(P <0.05), as compared with NS group [ (0.82 ± 1.20)μm, (0.42 ± 0.25)N/mm and (0.75 ± 0.64)%, all P < 0.05].Conclusions The short-term administration of NaF on rats in the growing period increases the bone formation and osteoblast activities of young rats and adult rats. The long-term administration of NaF on rats does not increase the bone formation of rats in growth period. The osteoblast activities as well as the bone absorption of lumbar vertebra were strengthened. The likelihood of bone fracture became larger. The negative effects on bone metabolism and bone quality of rats were gradually displayed along with the prolongation of sodium fluoride usage.     

中药复方护骨胶囊对去卵巢大鼠骨骼结构和骨重建状态的影响

目的观察护骨胶囊(Hugu capsules,HG)对去卵巢大鼠骨结构和骨重建状态的影响。方法 3月龄SPF级雌性SD大鼠60只采用数字表法随机分为假手术组,去卵巢组,仙灵骨葆胶囊组和护骨胶囊低、中、高剂量组(Sham、OVX、Pos、HG-L、HG-M和HG-H)。大鼠切除卵巢后,隔天开始给药,连续灌胃3个月,处死前进行两次荧光标记。取大鼠胫骨上段(proximal tibia,PTM)与中段(tibia shaft,TX)进行骨形态计量学分析。结果与Sham组比较:OVX组松质骨的骨小梁面积百分率(percent trabecular bone area,Tb.Ar)和骨小梁数量(trabecular number,Tb.N)分别降低66.68%和65.01%,骨小梁分离度(trabecular seperation,Tb.Sp)增加276.97%。单位骨小梁周长成骨细胞数(osteoblast number/unit trabecular perimeter,Ob.N)和成骨细胞周长百分率(percent osteoblast perimeter,Ob.Pm)分别增加357.53%和468.27%,单位骨小梁周长破骨细胞数(osteoclast number/unit trabecular perimeter,Oc.N)和破骨细胞周长百分率(percent osteoclast perimeter,Oc.Pm)分别增加213.95%和216.67%,骨形成率-体积(bone formation rate/volume,BFR/BV)增加39.12%,骨形成率-组织(bone formation rate/tissue,BFR/TV)降低55.56%,提示松质骨骨质疏松造模成功。皮质骨没有变化;与OVX组比:HG组大鼠松质骨的Tb.Ar、Tb.N和骨小梁宽度(trabecular width,Tb.Wi)增加,Tb.Sp、Oc.N、Oc.Pm降低,荧光周长百分率(percent fluorescence perimeter,L.Pm)、骨形成率-周长(bone formation rate/bone surface,BFR/BS)、BFR/BV和骨形成率-组织(bone formation rate/tissue,BFR/TV)增加。HG组大鼠皮质骨的T.Ar、Ct.Ar、骨外膜荧光标记周长百分率(percent periosteal fluorescence perimeter,P-L.Pm)和骨外膜骨形成率(periosteal bone formation rate/bone surface,P-BFR/BS)均增加。说明HG能增加松质骨骨量,改善骨结构,促进骨形成,抑制骨吸收,且增加皮质骨骨量,促进皮质骨骨外膜骨形成;与Pos组相比:HG-H组的松质骨Tb.N增加23.80%,Tb.Sp降低22.71%,HG-L和HG-M组的L.Pm、BFR/BS、BFR/BV和BFR/TV显著升高。在皮质骨方面,HG-L和HG-M组的P-L.Pm分别升高234.99%和182.58%,HG-L组的P-BFR/BS增加229.13%。结论 HG增加去卵巢大鼠的骨量,改善骨组织结构。HG具有促进6月龄OVX骨高转换大鼠骨形成的作用,在此年龄段促进骨形成方面效果可能优于Pos。为临床应用HG提供了良好的实验依据。     

近交系和封闭群Wistar大鼠糖皮质激素致骨质疏松造模比较

目的比较近交系和封闭群wistar大鼠在地塞米松(dexamethasone,Dex)不同给药频率作用下骨量、骨结构和骨力学性能变化,为不同遗传背景大鼠的糖皮质激素性骨质疏松(glucocorticoid-induced osteoporosis,GIOP)模型制备提供实验依据。方法 3月龄SPF级两种品系雌性Wistar大鼠各32只采用数字法随机分为4组:近交系的对照组(J-C)和Dex低、中、高频给药组(J-L、J-M和J-H);封闭群的对照组(F-C)和Dex低、中、高频给药组(F-L、F-M和F-H),C组肌注0.9%氯化钠注射液(1 m L/kg,3次/周);L、M和H组分别肌注Dex(1 mg/kg,分别1、3、5次/周),共给药3个月,处死前进行2次荧光标记。取大鼠胫骨上段(proximal tibia,PTM)进行骨组织形态计量学分析,取股骨和第五腰椎行分别行三点弯曲和压缩试验测定。结果(1)与J-C组比较,J-M和J-H组生长板下骨小梁结构紊乱,各给药组静态参数差异均无统计学意义(P0.05);J-H组骨矿化沉积率(mineral apposition rate,MAR)、荧光周长百分率(percent fluorescence perimeter,%L.Pm)、骨形成率-组织(bone formation rate/tissue,BFR/TV)、骨形成率-体积(bone formation rate/volume,BFR/BV)和骨形成率-周长(bone formation rate/bone surface,BFR/BS)分别下降39.27%、61.92%、79.21%、75.08%和77.14%;单位骨小梁周长成骨细胞数(osteoblast number/unit trabecular perimeter,Ob.N/BS)、成骨细胞周长百分率(percent osteoblast perimeter,%Ob.S/BS)分别下降57.35%和61.16%;单位骨小梁周长破骨细胞数(osteoclast number/unit trabecular perimeter,Oc.N/BS)、破骨细胞周长百分率(percent osteoclast perimeter,%Oc.S/BS)分别下降50.29%、42.92%;最大载荷、最大应力、弹性载荷、股骨长度、最大断裂吸收能、骨材料韧性均显著下降(P0.05)。(2)与F-C组比较,F-M组和F-H组均呈骨量减少、骨结构破坏表现,F-H组MAR、%L.Pm、BFR/TV、BFR/BV、BFR/BS差异均有统计学意义(P0.05);Ob.N/BS和%Ob.S/BS分别下降87.36%、87.9%,Oc.N/BS、%Oc.S/BS分别增加186%、123%;三点弯曲及压缩参数均显著下降。结论以3、5次/周肌注Dex 1 mg/kg给药3个月,可以成功诱导封闭群Wistar大鼠GIOP成模,但未能使近交系wistar大鼠呈现低骨量效应,显示不同遗传背景个体及不同部位骨组织对糖皮质激素骨损伤效应有易感性差异。相对于骨量指标,反映骨结构、骨强度和骨形成功能的指标对糖皮质激素骨损伤效应较为敏感。     

复方中药对慢性氟中毒大鼠氟骨症治疗效果的显微CT观察

目的 采用显微CT技术评价复方中药方剂对慢性氟中毒大鼠氟骨症的治疗效果.方法 断乳2周的纯系Wistar大鼠88只,体质量(91.1±10.0)g,按体质量采用随机数字表法分为对照组、中氟组、高氟组、高氟低钙低蛋白组,分别为16、24、24、24只大鼠.中氟组、高氟组、高氟低钙低蛋白组染氟剂量分别为50、100、100 mg/kg,高氟低钙低蛋白组饲料中蛋白质与钙的含量为中氟组和高氟组的1/2.染氟6个月后,每组采用股动脉放血法处死8只大鼠;3个染氟组剩余16只大鼠又分为两小组,一组为持续染氟对照组,另一组模拟氟中毒病区实际情况在持续染氟的基础上用复方中药进行治疗,每天每只大鼠按100 g体质量给药194 mg,每周灌服6d;分别于治疗前和治疗后30、60d收集大鼠24h尿样.大鼠连续灌服90 d,股动脉放血法处死大鼠,分离四肢骨.氟离子选择电极法检测大鼠尿氟;高温灰化-氟离子选择电极法检测骨氟;显微CT技术检测大鼠四肢骨的骨矿物质密度(BMD)、组织骨密度(TMD)、结构模型指数(SMI)、骨小梁厚度(Tb.Th)、骨小梁分离度(Tb.Sp)、各向异性(a1/a3)、骨小梁连接密度(Conn.D)、骨小梁与全部骨组织体积比(BV/TV)、骨表面积与体积比(BS/BV)、骨小梁数目(Tb.N).结果 复方中药治疗后60d,高氟低钙低蛋白治疗组尿氟[(11.01±3.67)mg/L]低于高氟低钙低蛋白对照组[(34.32±9.50)mg/L,t=3.13,P＜ 0.05].复方中药治疗后90d,高氟治疗组骨氟[(275.38±171.65) mg/kg]低于高氟对照组[(701.67±178.16)mg/kg,t=5.42,P＜ 0.05],高氟低钙低蛋白治疗组骨氟[(313.26±124.51)mg/kg]低于高氟低钙低蛋白对照组[(794.66±261.35) mg/kg,t=3.25,P＜0.05].复方中药治疗后90d,各组大鼠Tb.Th、Tb.Sp、a1/a3、Conn.D、BV/TV、BS/BV、Tb.N组间比较差异有统计学意义(F值分别为2.785、2.681、3.039、27.231、2.595、2.854、5.050,P均＜0.05).其中中氟治疗组大鼠Tb.Th、Tb.Sp[(0.04±0.01)、(0.03±0.01)mm]高于中氟对照组[(0.02±0.00)、(0.02±0.00)mm,P均＜0.05],a1/a3、Conn.D、BV/TV、Tb.N[(0.77±0.61),(510.91±304.99)mm-3,(0.42±0.06),(13.58±2.48) mm-1]低于中氟对照组[(1.11±0.01),(2 403.69±124.02)mm-3,(0.46±0.03),(18.12±0.69)mm-1,P均＜0.05];高氟治疗组大鼠BV/TV(0.44±0.04)低于高氟对照组(0.49±0.00,P＜0.05),Tb.Th[(0.04±0.01)mm]高于高氟对照组[(0.03±0.00)mm,P＜ 0.05].结论 复方中药对大鼠氟骨症存在一定的治疗效果.     

慢性氟中毒对大鼠精子活动度的影响

目的 探讨慢性氟中毒对大鼠精子活动度的影响,为氟的生殖毒性研究提供实验依据.方法 健康雄性Wistar大鼠32只,体质量150～180 g,按体质量随机分为4组:生理盐水(对照)组、低、中、高氟组(100、200、300mg·kg~(-1)·d~(-1)NaF),灌胃染毒90 d,每天称体质量.染毒结束后处死大鼠,摘取肝脏、肾脏、睾丸,称质量并计算脏器系数;取附睾游离精子,WLJY-9000型伟力彩色精子质量检测系统测定精子运动参数.结果 染毒第30天低、中氟组大鼠体质量[(235.00±14.56)、(235.44±24.99)g]高于高氟组[(206.00 ±18.16)g,P均<0.05)];第60、90天组大鼠体质量比较差异无统计学意义(F值分别为0.578、1.893,P均>0.05).大鼠肝脏系数、肾脏系数、睾丸系数组间比较差异无统计学意义(F值分别为2.148、0.907、1.801,P均>0.05).精子运动参数,平均路径速度(VAP)高氟组[(25.04 ±4.59)μm/s]较对照组[(20.22 ±3.29)μm/s]升高;直线速度(VSL)低、中、高氟组[(18.82 ±3.19)、(17.84 ±4.54)、(16.46 ±2.63)μm/s]较对照组[(12.48 ±1.73)μm/s]升高;直线性(LIN)低、中、高氟组[(23.84±1.58)%、(24.99±3.37)%、(26.75 ±5.07)%]较对照组[(33.29±4.00)%]降低,摆动性(WOB)中、高氟组[(47.03 ±3.98)%、(49.21±7.73)%]较对照组[(38.09 ±0.48)%]升高;平均移动角度(MAD)低氟组[(68.29 ±5.71)度/s]较对照组[(81.57 ±8.44)度/s]降低;鞭打频率(BCF)高氟组[(11.47 ±0.61)次/s]较对照组[(9.49 ±0.34)次/s]升高;精子密度(p)低、中氟组[(1.26 ±0.24)×10~9、(1.84 ±0.50)×10~9/L]较对照组[(3.94±1.10)×10~9个/L]降低(P均<0.05);曲线速度(VCL)、前向性(STR)、侧摆幅度(ALH),组间比较差异无统计学意义(F值分别为0.264、2.209、1.667,P均>0.05).结论 慢性氟中毒可影响大鼠精子活动度.降低精子密度.损害大鼠生殖系统.     

氟铝联合中毒小鼠骨骼病理与形态计量学研究

目的 探讨氟铝联合中毒小鼠骨相组织病变特征及致病的可能机制.方法 采用析因设计方法,将昆明种小鼠分成9组,由氟化钠(NaF,0、50、150 mg/L)、氯化铝(AlCl3,0、200、600mg/L)经饮水染毒,24周后对斑釉牙发生情况进行分度,测量股骨下端骨骺的骨小梁面积、类骨质面积,计数成骨细胞和破骨细胞个数,并观察骨组织病理变化.结果 铝单独作用也可引起斑釉牙(分度为4度),氟铝联合对牙的损害比各自单独作用严重.氟铝共存对骨小梁面积和类骨质面积的影响存在交互作用(F值分别为2.963、3.688,P＜0.05),对成骨细胞和破骨细胞数的影响不存在交互作用(F值分别为2.347、0.888,P＞0.05).高氟组骨小梁面积、类骨质面积为(50675.47±22 916.34)、(10 733.97±3015.55)μm2,高氟高铝组升高到(75 988.64±13 797.21)、(16 402.88±4605.83)μm2(P＜0.05),高氟低铝组升高到(69 277.16 ±19 837.51)、(18 564.79 4-6362.47)μm2(P＜0.05),铝拮抗了氟的效应;高铝组骨小梁面积为(60 718.43 4-17 574.37)μm2,高氟高铝组[(75 988.64 ±13 797.21)μm2]、低氟高铝组[(82 474.94±15 466.6)μm2]均升高(P＜0.05),联合作用效应与高剂量铝的作用趋势一致.骨组织病理改变为骨质疏松明显,类骨质和软骨组织增多,骨基质和矿化骨减少.结论 高剂量的铝和氟均可引起斑釉牙,联合作用比单一因素作用强.氟铝联合中毒对骨骼的影响以软化型骨质疏松为主要特征.     

氟铝联合对雄性大鼠性激素的影响

目的 了解氟铝联合对雄性大鼠性激素的影响.方法 选用断乳2周的健康SD雄性大鼠16只,按体质量随机分为4组,每组4只,分别为对照组及铝、氟、氟铝组.对照组和铝组喂饲的玉米饲料68%来自于非病区(含氟、铝各为5.2、6.8 mg/kg),分别给含铝0、90.0 mg/L的饮水;氟组、氟铝组给含68%的燃煤型氟中毒病区煤烘玉米饲料,含氟、铝量为106.0、19.7 mg/kg,并分别给予含铝0、90.0 mg/L的饮水.实验第90天后以出现明显氟斑牙为模型复制成功的判定指标.采集大鼠血样,用时间分辨免疫荧光法进行血清中睾酮(T)、雌二醇(E2)的测定.结果 大鼠血清T水平氟铝组[(15.994±6.558)μg/L]明显高于对照组[(3.317±0.635)μg/L],差异有统计学意义(P＜0.05),铝组[(8.134±3.134)μg/L]、氟组[(1.868±0.367)μg/L]、氟铝组[(12.687±2.979)μg/L]与对照组比较,差异无统计学意义(P＞0.05).大鼠血清E2氟组[(0.172±0.030)nmol/L]明显低于对照组[(0.319±0.072)nmol/L],差异有统计学意义(P＜0.05),铝组[(0.282±0.012)nmol/L]、氟铝组[(0.265±0.047)nmol/L]与对照组比较,差异无统计学意义(P＞0.05).氟和铝两因素存在交互作用(F=9.82,P＜0.05).结论 氟铝联合作用影响雄性大鼠性激素的水平.     

铁蓄积骨质疏松小鼠模型造血干细胞与骨代谢指标的相关性

目的了解铁蓄积骨质疏松小鼠模型造血干细胞(hematopoietic stem cells,HSCs)指标变化及其与骨代谢指标变化的相关性,探讨骨质疏松范畴"铁蓄积"对造血系统的影响。方法 6~8周C57雄性小鼠分为对照组、实验组,实验组用枸橼酸铁铵(ferric ammonium citrate,FAC)0.1 g/kg干预8周,两组均检测:血清铁蛋白(ferritin,FER)、胶原1型氨基端前肽(procollagen type 1 N-terminal propeptide,P1NP)、破骨标志物(carboxy-terminal telopeptides,CTX)、外周血血常规、股骨远端骨小梁三维形态重建和空间结构参数、HSCs占骨髓细胞比例、HSCs细胞集落形成能力,统计分析两组实验数据。结果实验组骨髓HSCs占骨髓比例4.01%±0.63%较对照组2.61%±0.43%显著升高(P0.05),实验组HSCs细胞集落形成142个/皿较对照组47个/皿显著增多;实验组血清铁蛋白(13.27±0.72)μg/L较对照组(4.14±0.96)μg/L显著升高,实验组P1NP(4.62±0.76)ng/ml较对照组(8.23±0.66)ng/ml显著降低,实验组CTX(123.54±1.63)ng/ml较对照组(83.36±1.89)ng/ml显著升高(P0.05);实验组组外周血血小板(1380±103)×109/L较对照组(1023±83)×109/L显著升高(P0.05);实验组micro-CT检测的骨密度(50.41±10.27)mg/mm~3较对照组(104.68±8.56)mg/mm3显著下降、实验组骨小梁空间结构参数[bone volume/tissue volume,BV/TV;骨体积分数10.18%±1.76%;trabecula thickness,Tb.Th骨小梁厚度(0.057±0.012)mm;trabecula number,Tb.N骨小梁数(1.02±0.21)N/mm]较对照组[BV/TV(19.92%±1.92%);Tb.Th(0.098±0.008)mm;Tb.N(1.45±0.13)N/mm]显著下降(P0.05)。结论在铁蓄积影响的骨质疏松模型中,铁蓄积不仅对骨代谢指标有显著影响,也对HSCs有显著影响。     

染氟小鼠成骨细胞系MC3T3-E1细胞中还原型谷胱甘肽和氧化型谷胱甘肽观察

目的 观察染氟小鼠成骨细胞系MC3T3-E1细胞中还原型谷胱甘肽(GSH)和氧化型谷胱甘肽(GSSG)的水平.方法 采用噻唑蓝(MTT)法检测不同染氟条件下[F-剂量分别为0(对照)、0.5、1.0、2.0、4.0、8.0、12.0、20.0 mg/L]MC3T3-E1细胞1、2、4、10 d的细胞活性.另外,按染氟剂量,将MC3T3-E1细胞分为0(对照)、2、8、20 mg/L组,分别染氟2、4、10 d,采用高效液相色谱—串联四极杆质谱技术检测细胞内GSH、GSSG和谷氨酰胺(Gln)水平.结果 染氟1d时2.0 mg/L组细胞活性(0.57±0.05)明显高于对照组(0.49±0.03,P＜0.01);染氟4d时8.0、12.0 mg/L组细胞活性(0.49±0.07、0.47±0.09)明显低于对照组(0.63±0.06,P＜ 0.05或＜ 0.01);染氟10d时8.0 mg/L组细胞活性(1.52±0.29)明显高于对照组(0.86±0.23),而20.0 mg/L组细胞活性(0,54±0.07)明显低于对照组(P均＜0.01).染氟2、10d时20 mg/L组细胞中GSH水平[(13.92±4.63)、(0.53±0.30) μmol/L]明显低于相应的对照组[(26.42±3.67)、(24.85±5.68)μmol/L,P均＜0.01].染氟2d时2 mg/L组、染氟4d时8 mg/L组、染氟10 d时8 mg/L组细胞内GSSG水平[(1.12±0.62)、(2.13±0.62)、(2.97±1.30)μmol/L]明显高于相应的对照组[(0.55±0.22)、(1.46±0.46)、(1.35±0.50)μmol/L,P＜ 0.05或＜ 0.01].染氟4d时2 mg/L组和染氟10d时8、20 mg/L组细胞内Gln水平[(62.80±17.41)、(122.26±19.51)、(19.38±8.11) μmol/L]明显低于相应的对照组[(83.28±14.32)、(147.15±16.95) rmol/L,P均＜0.05或＜ 0.01].结论 染氟能明显改变成骨细胞内的GSH、GSSG和Gln水平,从而影响细胞内氧化还原平衡态.     

Bone histomorphometry in 50 normal tunisian subjects

Summary Quantitative bone histomorphometry was done on undecalcified sections of iliac crest bone specimens obtained at autopsy from 50 normal subjects (24 males and 26 females). The following parameters were measured: cortical thickness (Ct.Th), trabecular bone volume (BV/TV), trabecular number (Tb.N), trabecular thickness (Tb.Th), osteoid volume (OV/BV), osteoid surfaces (OS/BS), osteoid thickness (O.Th) and eroded surfaces (ES/BS). There was a significant age-related decrease in BV/TV in both sexes which followed a x³ polynomial regression. A significant decrease of Tb.Th was noted in males after the fifth decade. In males, bone loss was 1.5% per decade, but in females it was 0.36% before menopausal period and 2% after. Other parameters were unrelated to age and sex.     

Effect of Sevelamer Hydrochloride on Bone in Experimental Uremic Rats

Abstract: Hyperphosphatemia in dialysis patients is known to cause secondary hyperparathyroidism and high-turnover bone disease. Sevelamer hydrochloride (sevelamer) is a nonabsorbed, calcium-free phosphate-binder. We determined the effect of sevelamer on parathyroid hormone (PTH)-induced high bone turnover. Rats were sham-operated or 5/6-nephrectomized (Nx) and fed a phosphate loading diet for 16 weeks or 5/6-nephrectomized and fed a phosphate loading diet for 8 weeks and then fed the same diet containing 3% sevelamer for the subsequent 8 weeks (Nx-S). Sevelamer significantly reduced serum PTH. The relative osteoid volume (OV/BV), osteoid surface (OS/BS), eroded surface (ES/BS), mineral appositional rate (MAR), volume-referent bone formation rate (BFR/TV), and bone-referent bone formation rate (BFR/BV) were measured for vertebral bone histomorphometric analysis. All parameters were statistically higher in the Nx rats than in the sham-operated control rats. The administration of sevelamer attenuated increases in OV/BV, ES/BS, BFR/TV, and BFR/BV. For femur histomorphometric analysis, the porosity area (%) (PoAr/CtAr), osteoid surface on the periosteal surface, osteoid surface on the endocortical surface (OS/Es), mineral appositional rate on the periosteal surface, mineral appositional rate on the endocortical surface, bone formation rate on the periosteal surface, and bone formation rate on the endocortical surface (Es BFR) were calculated. All parameters were higher in the Nx group than in the control group. Sevelamer inhibited the elevation of PoAr/CtAr, OS/Es, and Es BFR. Our findings suggest that the decrease in PTH by sevelamer may be beneficial in the treatment of high PTH-induced bone disease.     

仙灵骨葆对去势大鼠骨量和生物力学性能的影响

目的探讨仙灵骨葆对骨质疏松(OP)大鼠骨量、骨代谢和生物力学性能的影响。方法 3月龄雌性SD大鼠24只分为3组,每组8只:正常对照组(N)、卵巢切除组(OVX)、卵巢切除+仙灵骨葆治疗组(XLGB)。除N组外,其余两组行卵巢切除术,6 w后XLGB组给予药物干预:250 mg.kg-1.d-1,OVX组给予等量生理盐水,8 w后处死所有大鼠。留取尿液、血清检测血PINP值、尿DPYD/Cr、NTX/Cr值。取左侧股骨行骨密度测定,取左侧胫骨制备硬组织不脱钙切片,备行骨组织形态计量学检测,取右侧股骨行三点弯曲试验,检测其最大载荷。结果 OVX组血PINP、尿DPYD/Cr、尿NTX/Cr值显著高于N组,XLGB能显著降低血PINP、尿DPYD/Cr、尿NTX/Cr值,但仍显著高于N组。OVX组股骨全长及近、中、远三段骨密度均显著低于N组,XLGB组近、远端骨密度显著高于OVX组。BV/TV在OVX组显著低于N组,XLGB组显著高于OVX组;OVX、XLGB组骨吸收指标Oc.N、Er.Pm均显著高于N组,XLGB组Oc.N、Er.Pm显著低于OVX组,BFR/BV显著高于OVX组。最大载荷三组之间无显著差别。结论仙灵骨葆灌胃可抑制卵巢切除大鼠骨量丢失,其机制与促进骨形成、抑制骨吸收,降低骨转换水平,进而维持骨量及微观结构有关。     

去卵巢大鼠椎体松质骨骨小梁微结构改变及其对生物力学的影响

目的观察骨质疏松和正常状态下椎体松质骨的微观结构改变,分析其对骨生物力学的影响。方法将12只4月龄雌性Lewis大鼠随机分为去卵巢组(OVX)组和假手术组(Sham),每组各6只。OVX组行双侧卵巢切除术,假手术组仅显露双侧卵巢。术后6个月处死动物,取尾椎(L_(4-7))行Micro-CT分析及生物力学测试。结果去卵巢6个月后,OVX组大鼠体积骨密度(vBMD)和组织骨密度(tBMD)较Sham组显著降低,松质骨骨小梁的骨体积分数(BV/TV)和数目(TB.N)都明显低于Sham组,骨小粱表面积密度(BS/BV)、结构模拟指数(SMI)和间距(Tb.Sp)显著高于Sham组,差异有统计学意义。但2组间骨小梁厚度(Tb.Th)差异无统计学意义。生物力学测试结果表明,去势6个月后,OVX组大鼠骨质生物力学性能显著下降。骨小梁力学性能与骨小梁体积分数(Adjusted R~2=0.750和数目(Adjusted R~2=0.861)呈正线性相关,而与结构模拟指数(Adjusted R~2=0.716)和骨小梁间距(Adjusted R~2=0.830)呈负线性相关。结论松质骨骨小梁微观结构的改变可影响骨质的生物力学性能,二者之间具有一定的线性关系。     

旋转中心偏移对显微CT松质骨定量分析的影响

目的观察旋转中心微小偏移对显微CT测量参数的影响。方法20只7月龄sD大鼠随机分为去卵巢组（OVX）和假手术组（SHAM）。手术后3周处死,应用显微cT扫描胫骨近干骺端。手动校正以获取每次扫描的最佳旋转中心,分析各旋转中心在偏移±0．5、±1．0、±1．5和±2．0像素条件下的密度及微结构参数。结果一般线性模型分析结果显示OVX组与SHAM组比较,表观骨密度、组织骨密度、骨体积分数、骨小梁数量和联接密度明显下降,骨小梁间隔明显增宽（P〈0．05）。组织骨密度、各向异性度、骨小梁面积密度和联接密度随旋转中心的偏移下降,骨体积分数和骨小梁厚度随偏移幅度的加大逐渐升高。旋转中心偏移±1．5像素内,对组织骨密度、骨体积分数、各向异性度和联接密度的测量无影响。而骨小梁厚度和骨小梁面积密度在旋转中心偏移±1．0像素内影响较小。OVX组与SHAM组各参数随旋转中心偏移的变化趋势基本一致。各参数随旋转中心偏移服从二次回归方程趋势。通过二次回归方程拟合,可以获得实际旋转中心。以该中心获取的图像质量高,并能确保定量分析数据的准确。实际旋转中心与人为校正的最佳旋转中心之间存在微小差异。结论旋转中心的微小偏移对表观骨密度、结构模型指数、骨小梁间隔和数量的测量无明显影响。组织骨密度、骨小梁厚度、骨小梁面积密度、骨体积分数、各向异性度和联接密度受旋转中心偏移影响较大。通过二次曲线方程拟合能找到正确的旋转中心,该中心与人为校正获取的最佳旋转中心存在一定误差。     

大鼠去卵巢后不同时期骨微结构和力学性能的变化特征

目的揭示大鼠在去卵巢后不同时期腰椎松质骨微结构退变的变化特征,探讨骨整体力学性能下降的同时可能存在的各种适应性代偿性变化。方法50只7月龄SD大鼠随机分为基线、去卵巢组（OVX组）和假手术组（SHAM）。基线组10只,其余每组均20只。实验开始时先将基线组10只处死,OVX组和SHAM组分别在手术后3周、15周各处死10只,留取动脉血清及腰椎标本,骨微结构、力学和生化指标的测定。结果去卵巢后3周时OVX组表观骨密度、骨体积分数、骨小梁厚度和骨小梁数量均较SHAM和基线降低（P〈0．05）,各向异性度较基线下降（P〈0．05）而与SHAM组无统计学差异。骨小梁面积密度、骨小梁间隔和结构模型指数均较SHAM和基线组增加（P〈0．05）。去卵巢后3周时OVX组最大应力、弹性模量、血清TRAP-5b和骨细胞密度均低于基线（P〈0．05）而与SHAM组无统计学差异。去卵巢后15周时OVX组表观骨密度、骨体积分数、骨小梁数量和联接密度、最大应力、TRAP-Sb和骨细胞密度均较SHAM和基线组降低（P〈0.05）,骨小梁结构模型指数、骨小梁间隔和各向异性度均较SHAM和基线组增加（P〈0．05）,骨小梁面积密度和厚度均与SHAM和基线组无统计学差异。结论大鼠去卵巢后腰椎骨量快速丢失,骨微结构逐渐退变,而血清TRAP-5b水平下降及骨细胞密度、骨小梁各向异性度和厚度的适应性增加,可能在一定程度上代偿骨力学性能的下降,有利于维持骨结构的完整性。     

复方淫羊藿对去卵巢大鼠腰椎松质骨的影响

目的 观察淫羊藿提取物与乙烯雌酚（DES）组成复方淫羊藿对去卵巢大鼠腰椎松质骨的影响。方法 4.5月龄SD雌性大鼠70只，随机分成假手术组、去卵巢组、DES组、淫羊藿提取物组、复方淫羊藿高、中、低剂量组，持续给药90d。大鼠处死后用骨组织形态计量学等方法测量腰椎松质骨的动态参数和静态参数，同时测量子宫内膜厚度。结果 复方淫羊藿3个剂量组的腰椎％TbAr、Tb.Th均高于去卵巢组，而Tb.Sp、Oe.N、BFR／BV均低于去卵巢组。复方淫羊藿商剂量组的作用比单用DES强，而低剂量组的作用与DES相当。复方淫羊藿3个剂量组子宫重量、子宫内膜厚度均高于去卵巢组，但子宫内膜厚度均小予DES组。结论 复方淫羊藿预防去卵巢大鼠腰椎松质骨丢失的作用强于DES，对子宫刺激的作用比DES小。     

Magnetic resonance imaging of normal and osteoarthritic trabecular bone structure in the human knee

OBJECTIVE: To use high-resolution magnetic resonance imaging (MRI) to evaluate the trabecular bone structure in the distal femur and the proximal tibia and its to correlate the findings with different stages of osteoarthritis (OA) of the human knee. METHODS: Axial images of the distal femur and proximal tibia were obtained at 1.5 T in patients without and with mild OA and with severe OA. The spatial resolution was 195 x 195 microm(2) with a 1-mm slice thickness. Apparent measures of trabecular bone volume fraction (BV/TV), trabecular number (Tb.N), trabecular separation (Tb.Sp), and trabecular thickness (Tb.Th) were calculated. RESULTS: Significant differences existed in the trabecular bone structure of the femur and tibia. Differences in trabecular bone structure between the tibia and the femur decreased with the degree of OA. The apparent BV/TV, Tb.N, and Tb.Sp in the femoral condyles could be used to differentiate healthy patients or patients with mild OA from patients with severe OA (P < 0.05). Among individuals, the structural variation of the lateral and medial femoral condyle was indicative of the extent of the disease. CONCLUSION: High-resolution MRI of the knee joint can provide a noninvasive assessment of trabecular bone structure. Trabecular bone structure, determined by high-resolution MRI, shows significant variation in patients with varying degrees of OA. The impact of OA on trabecular bone is different in the tibia than in the femur, and this difference depends on the extent of the disease.     

骨细胞密度对骨生物力学性能的影响

目的 探讨骨细胞密度是否为衡量骨生物力学性能的指标. 方法 40只7月龄雌性SD大鼠随机分为去卵巢组(OVX)、OVX+金雀异黄酮(GEN,5 mg·kg-1·d-1)和OVX+1713雌二醇(EST,10 μg·kg-1·d-1)组及假手术组(SHAM),每组lO只.手术后15周第5腰椎(L5)椎体进行压缩试验,L6椎体先行显微CT扫描测量三维骨密度和骨微结构参数,然后进行疲劳损伤试验,最后行大块组织品红染色、塑料包埋和磨片.磨片标本用于微损伤、骨细胞密度检测. 结果 去卵巢后15周时,OVX组骨细胞密度为(1268.1±191.2)个/mm2,较SHAM组(1760.8±376.6)个/mm2及EST组(1550.9±202.2)个/mm2降低(F=3.531,P<0.05);OVX组最大应力为(84.4±16.9)N,较SHAM(110.3±25.6)N、EST(103.9±15.8)N及GEN组(110.1±4.9)N降低(F=9.561,P<0.05);OVX组骨小梁连接密度为(47.4±7.4)mm-3,较SHAM(71.8±16.0)mm-3及EST组(74.0±12.7)mm-3降低(F=7.635,P<0.05);OVX组骨矿含量为(6.5±2.2)g,较SHAM组(7.9±1.2)g降低(F=2.489,P<0.05);OVX组微破裂平均长度(58.1±6.8)μm,较SHAM(24.2±8.1)/Lm、EST(36.5±9.7)μm及GEN组(28.5±7.5)μm增加(F=3.179,P<0.05);OVX组骨小梁间隔(315.0±32.7)μm,较SHAM(222.5±21.7)μm及EST组(273.3±50.O)μm增加(F=7.007,P<0.05).骨细胞密度与最大应力(R2=0.7874,P<0.05)、骨小梁连接密度(R2=0.1153,P<0.05)、骨矿含量(R2=0.1309,P<0.05)呈正相关,与微破裂平均长度(R2=0.5738,P<0.05)、骨小梁分离度(R2=0.3964,P<0.05)负相关. 结论 骨细胞在维持骨力学性能中起重要作用,骨细胞密度可能是潜在的评价骨力学性能的重要指标.     

骨细胞GLP-1受体表达对2型糖尿病大鼠骨微结构的影响

目的观察2型糖尿病大鼠股骨颈骨细胞表达胰高血糖素样肽-1受体(GLP-1R)阳性细胞百分数的变化,探讨骨组织GLP-1R变化对骨微结构的影响。方法雄性Wistar大鼠60只,随机分为实验组(30只)和对照组(30只),实验组采用高糖高脂饲料喂养至12周时加小剂量链脲佐菌素腹腔注射诱导糖尿病模型,对照组采用普通饲料喂养。分别于造模成功时和成模后第12周,处死10只大鼠作为糖尿病0周组(DM0w组,n=10)和糖尿病12周组(DM12w组,n=10),对照组分别于同期各处死10只大鼠作为正常对照(NGT0w组,n=10;NGT12w组,n=10)。经腹主动脉采血测定空腹血糖(FBG)、胆固醇(TC)、三酰甘油(TG)。取近端股骨颈,制备脱钙和不脱钙骨切片,备免疫组化检测GLP-1R阳性的骨细胞,采用Simple软件分析骨小梁形态计量学指标:骨小梁占骨髓腔体积百分比(BV/TV)、骨小梁厚度(Tb.Th)、骨小梁数目(Tb.N)。采用单因素方差分析比较各组大鼠股骨颈表达GLP-1R阳性的骨细胞百分数,BV/TV、Tb.Th、Tb.N的变化,采用Pearson行相关分析。结果与同期对照组相比,DM0w组、DM12w组大鼠FBG、TC、TG升高(均P0.05);与同期对照组相比,DM0w组、DM12w组大鼠股骨颈表达GLP-1R阳性骨细胞百分数和BV/TV、Tb.Th均减少(均P0.05);与DM0组相比,DM12组大鼠股骨颈表达GLP-1R阳性的骨细胞百分数、BV/TV、Tb.Th、Tb.N均减少(均P0.05);Pearson相关分析显示大鼠股骨颈GLP-1R阳性骨细胞百分数与股骨颈BV/TV、Tb.Th、Tb.N均呈正相关,相关系数分别为r=0.803,P0.05;r=0.545,P0.05;r=0.771,P0.05。结论糖尿病大鼠股骨颈骨细胞GLP-1R的表达减少,同时BV/TV、Tb.N减少,Tb.Th变薄,骨细胞GLP-1R的表达可能是影响2型糖尿病骨微结构的因素之一。