CAJ | 학술논문

目的探讨吡咯并喹啉醌(pyrroloquinoline quinone,PQQ)对Bmi-1基因敲除(Bmi-1knock out,BKO)小鼠的下颌骨及牙发育障碍的治疗作用及其机制.方法将同窝的Bmi-1+/-雌雄小鼠进行配对,取7周龄正常饮食WT小鼠(10只,WT组),正常饮食BKO小鼠(10只,BKO组),以及正常饮食添加PQQ的BKO小鼠(10只,BKO+PQQ组),利用X线和显微CT、HE染色、组织化学及免疫组织化学、流式细胞仪检测等方法对3组小鼠分别检测下颌骨及牙的大小和骨密度、下颌牙槽骨骨皮质厚度、下颌第一磨牙前期牙本质厚度、下颌骨成骨及破骨细胞数、股骨、胸腺和肝脏的活性氧类水平等指标,并对计量资料结果3组间的总体比较和两两比较分别行单因素方差分析和t检验.结果 WT组小鼠表型正常,BKO+PQQ组小鼠整体表型较BKO组小鼠部分改善、体质量增加、生存期明显延长;X线及显微CT结果均显示BKO+PQQ组小鼠下颌骨及牙的大小和骨密度增加;BKO+PQQ组小鼠下颌牙槽骨骨皮质厚度[(68.65±0.25) μm]与BKO组小鼠[(42.45±0.35) μm]相比差异有统计学意义(P＜0.01);BKO+PQQ组小鼠下颌第一磨牙前期牙本质厚度[(4.25±0.15)μm]与BKO组小鼠[(31.55±0.35) μm]相比差异有统计学意义(P＜0.001);PQQ组小鼠下颌骨成骨细胞数[(38.45±0.25)个/mm3]显著高于BKO组小鼠[(18.15±0.55)个/mm3] (P＜0.01);BKO+PQQ组小鼠下颌骨破骨细胞数[(9.45±0.25)个/mm3]显著低于BKO组小鼠[(14.25±0.35)个/mm3] (P＜0.01);与BKO组小鼠相比,BKO+PQQ组小鼠股骨、胸腺和肝脏的活性氧类水平均显著下降(P＜0.01).结论 PQQ通过促进成骨细胞骨形成、减少破骨细胞骨吸收、清除活性氧类、减少DNA损伤等综合作用发挥对BKO小鼠牙及下颌骨发育障碍的治疗作用.
목적 탐토필각병규람곤(pyrroloquinoline quinone,PQQ)대Bmi-1기인고제(Bmi-1knock out,BKO)소서적하합골급아발육장애적치료작용급기궤제.방법 장동와적Bmi-1+/-자웅소서진행배대,취7주령정상음식WT소서(10지,WT조),정상음식BKO소서(10지,BKO조),이급정상음식첨가PQQ적BKO소서(10지,BKO+PQQ조),이용X선화현미CT、HE염색、조직화학급면역조직화학、류식세포의검측등방법대3조소서분별검측하합골급아적대소화골밀도、하합아조골골피질후도、하합제일마아전기아본질후도、하합골성골급파골세포수、고골、흉선화간장적활성양류수평등지표,병대계량자료결과3조간적총체비교화량량비교분별행단인소방차분석화t검험.결과 WT조소서표형정상,BKO+PQQ조소서정체표형교BKO조소서부분개선、체질량증가、생존기명현연장;X선급현미CT결과균현시BKO+PQQ조소서하합골급아적대소화골밀도증가;BKO+PQQ조소서하합아조골골피질후도[(68.65±0.25) μm]여BKO조소서[(42.45±0.35) μm]상비차이유통계학의의(P＜0.01);BKO+PQQ조소서하합제일마아전기아본질후도[(4.25±0.15)μm]여BKO조소서[(31.55±0.35) μm]상비차이유통계학의의(P＜0.001);PQQ조소서하합골성골세포수[(38.45±0.25)개/mm3]현저고우BKO조소서[(18.15±0.55)개/mm3] (P＜0.01);BKO+PQQ조소서하합골파골세포수[(9.45±0.25)개/mm3]현저저우BKO조소서[(14.25±0.35)개/mm3] (P＜0.01);여BKO조소서상비,BKO+PQQ조소서고골、흉선화간장적활성양류수평균현저하강(P＜0.01).결론 PQQ통과촉진성골세포골형성、감소파골세포골흡수、청제활성양류、감소DNA손상등종합작용발휘대BKO소서아급하합골발육장애적치료작용.
Objective To investigate the treatment effects and mechanisms of pyrroloquinoline quinine(PQQ) on defective teeth and mandible in Bmi-1 knockout mice.Methods Male and female Bmil+/-mice were paired with each other from the same nest.At the age of 7 weeks,the mice were divided into three groups,the wild type mice received normal diet(10 mice,WT group),Bmil-/-mice received normal diet (10 mice,BKO group),and the Bmi1+/-mice received normal diet and PQQ diet(10 mice,BKO+PQQ group).X-ray and micro-CT were used to detect mandible and dental size and bone mineral density.HE staining,histochemical and immunohistochemical methods were respectively used to detect alveolar bone thickness of cortical bone,predentin thickness of mandibular first molar,mandibular osteoblast number and osteoclast number.Flow cytometry was used to detect reactive oxygen species(ROS) levels of various organs(femur,thymus and liver).The data were statistically analyzed with one-way ANOVA and t test.Results Compared with BKO mice,BKO+PQQ mice partially rescued total body phenotype,increased body weight and prolonged survival time.X-ray and micro-CT showed the size of the mandible and teeth and bone mineral density of PQQ+BKO mice increased compared with BKO mice.In PQQ+BKO mice,mandibular alveolar bone cortical thickness[(68.65±0.25) μm] was significantly different from that in BKO mice[(42.45±0.35) μm] (P＜0.01).There was significant difference in predentin thickness of mandibular first molar between PQQ + BKO mice[(4.25 ± 0.15) μm] and BKO mice[(31.55 ± 0.35) μm] (P＜0.001).The number of osteoblasts in the mandible of BKO+PQQ mice[(38.45±0.25) cell/mm3] was significantly higher than that in the BKO mice[(18.15±0.55) cell/mm3] (P＜0.01).However,the number of osteoclasts in the BKO+PQQ mice [(9.45±0.25) cell/mm3] was significantly lower than that in the BKO group[(14.25±0.35) cell/mm3] (P＜0.01).Compared with the BKO group,ROS levels of the femur,thymus and liver in the BKO + PQQ mice were significantly decreased(P＜0.01).Conclusions The results indicate that PQQ may have treatment effects on defective teeth and mandible through promoting osteoblast bone formation and reducing osteoclast bone resorption,scavenging ROS and reducing DNA damage.