低能量激光联合压力对成骨细胞ALP活性和胞内Ca^(2+)浓度的影响  

Effect of LLLI on Secretion of ALP and Intracellular Calcium Concentration in Osteoblasts under Mechanical Stress

作　　者：史瑞新[1] 付喜宏[2] 庄金良[1] 杨陆一 

机构地区：[1]吉林大学口腔医学院正畸科,吉林长春130021 [2]中国科学院长春光学精密机械与物理研究所,吉林长春130033

出　　处：《光谱学与光谱分析》2016年第7期2178-2182,共5页Spectroscopy and Spectral Analysis

基　　金：国家自然科学基金项目(61405190);吉林省卫计委项目(2013zc007)资助

摘　　要：探讨低能量激光照射(1ow--level laser irradiation,LLLI)联合压力刺激对人成骨样细胞的碱性磷酸酶(alkaline phosphatase,ALP)活性和胞内Xa^(2+)浓度的变化规律的影响,揭示LLLI促进正畸牙齿压力侧骨改建的机制。将对数生长期的人成骨样细胞MG-63随机分为4组,对细胞施加压力采用Foreel四点弯曲体外细胞力学加载装置,用Ga-Al-As半导体激光器(808 nm,500 mW)进行照射。组Ⅰ(对照组):不加力,不进行激光照射。组Ⅱ(压力组):利用细胞加载装置对细胞加压力,加力板变形量为3 000 ustrain,加力时间1h。组Ⅲ(激光组):激光照射1 min,剂量3.75 J·cm。组Ⅳ(联合组):细胞加力(加力方式同组Ⅱ)后,激光照射(方式同组Ⅲ)。四组细胞均用分光光度计测量细胞内ALP活性。第二部分实验对胞内Ca^(2+)浓度变化进行测定,将MG-63细胞分为2组:压力组和激光张力组,2组细胞分别加力0,5,15,30,60 min,激光压力组再激光照射1 min后收取细胞。用荧光探针Fluo-3-AM测定成骨样细胞内Xa^(2+)浓度。结果显示第一部分实验与对照组相比,其余3组的ALP活性均明显增强(p<0.05),但联合组的ALP活性分别低于压力组和激光组(p<0.05)。第二部分实验显示:第1组压力引起胞内Ca^(2+)浓度随时间剧烈变化,第2组变化曲线平缓,但Xa^(2+)浓度水平两组无明显差异。由此得出结论适宜的压力和弱激光及联合应用均能增加成骨细胞的ALP活性,两者联合应用时较单独应用时成骨细胞的ALP活性稍有降低,压力组和激光压力组胞内Ca^(2+)浓度的变化曲线不同,说明在正畸牙齿的压力侧,低能量激光的应用可以降低由压力引起的成骨细胞活性增加,减少成骨,破骨相对增加,促进牙齿移动,LLLI极可能通过调节Xa^(2+)浓度的变化规律来调节成骨细胞活性。In order to reveal the mechanism of LLLI accelerating teeth moving ,we investigated the changes of alkaline phospha-tase and intracellular calcium concentration when osteoblasts under stress were subjected low-level-laser-irradiation （LLLI） . MG-63 cells were divided into four groups ：control group ,stress group ,LLLI group and LLLI-stress group .Osteoblasts were subjected to the mechanical stress by a four-point bending system at 0.5 Hz and 3 000 μstrain .The secretions of ALP of each group are measured by spectrophotometer .In the second part ,MG-63 cells were divided into two groups ：stress group and LL-LI-stress group .We checked intracellular calcium concentration via FCM and fluorescent indicator fluo-3/AM at 0 ,5 ,15 ,30 and 60 min under stress .LLLI-stress group will receive LLLI for 1 min after stress .Compared to a control group ,increased ALP secretions were observed in the other three groups .But ALP secretions in LLLI-stress group were lower than stress group and LLLI group .T HE changing curve of intracellular calcium concentration in laser-stress groups is gentle instead of “jumping”in stress group .Proper stress ,LLLI and combined application of these two can increase the secretions of ALP in osteoblasts compared to the control group .But the secretions of ALP decreased when combined application of stress and LLLI compared to using alone .LLLI can regulate the changing rhythm of concentration of the intracellular calcium to promote proliferation of MG-63 cell under stress ,which means LLLI can reduce the bone-formation of osteoblasts under stress .

关 键 词：低能量激光照射(LLLI) 成骨细胞 压力 碱性磷酸酶 

分 类 号：R318.5[医药卫生—生物医学工程]