810nm弱激光照射对大鼠急性脊髓损伤后巨噬细胞极化的作用研究  

The Effect of Low Level Laser Therapy on Macrophage Polarization after Acute Spinal Cord Injury In Rats

作　　者：宋基伟[1] 梁卓文[1] 李鲲[1] 沈学锋[2] 胡学昱[1] 王哲[1] 

机构地区：[1]第四军医大学西京骨科医院,陕西西安710032 [2]第四军医大学军事预防医学系劳动与环境卫生学教研室,陕西西安710032

出　　处：《激光生物学报》2017年第1期17-23,29,共8页Acta Laser Biology Sinica

基　　金：国家自然科学基金(81070996;81572151);陕西省社发攻关基金(2016SF-143)

摘　　要：本研究构建急性大鼠脊髓夹伤模型,并将大鼠随机分为单纯脊髓损伤对照组及脊髓损伤联合弱激光照射组。照射组应用810 nm波长,150 m W照射功率,照射光斑0.3 cm^2的弱激光对脊髓损伤区进行经皮照射,连续照射3天,7天或14天。应用免疫荧光、免疫印迹实验方法,测定脊髓损伤区巨噬细胞及小胶质细胞的极化表达。应用酶联免疫吸附法测定脊髓损伤区白细胞介素4的表达情况。应用坚牢蓝髓鞘染色测定两组损伤脊髓中髓鞘保留的差异。采用BBB评分对两组大鼠后肢运动功能的恢复进行评估。结果表明,810 nm弱激光对脊髓损伤区连续照射3天,7天后,可显著减少M1型巨噬细胞及其标志物诱导型一氧化氮合酶的表达,在7天时间增加M2型巨噬细胞及其标志物精氨酸酶1的表达。弱激光照射组白细胞介素4的表达明显增加。损伤后14天,弱激光照射组脊髓损伤区髓鞘保留面积比值明显提高。损伤后7天及14天时,弱激光照射组大鼠的BBB评分明显升高。该实验结果表明,810 nm弱激光经皮照射,可增加大鼠急性脊髓损伤区M2型巨噬细胞及小胶质细胞的表达,并减少脊髓损伤后的髓鞘脱失,促进脊髓损伤大鼠运动功能的恢复。The present study was designed to establish a bilateral compression spinal cord injury SCI rat model. 42 SD rats were randomly assigned to SCI only group （control group） or SCI followed by low level laser therapy group （ LLLT group）. After SCI, for the LLLT group, rats were received a laser beam （810 nm wavelength, 150 mW output power, 0. 3 cm^2 light spot） in percutaneous method once daily for consecutive 3, 7 or 14 days. Immunofluorescence （IF） and western blot （ WB） were performed to evaluate the polarization state of macrophage/microglia in the injured spinal cord. Meanwhile, enzyme linked immunosorbent assay （ELISA） was performed to assess the expression of interleukin 4 （IL- 4） and luxol fast blue （LFB） staining was used to assess the preservation of myelin sheath. Results of IF and WB re-vealed that Ml subset of macrophage/microglia and the cell marker inducible nitric oxide synthase（INOS） were signifi-cantly reduced in LLLT group at 3 and 7 days post injury （ dpi） . M2 subset and expression of the cell marker “arginasel （Argl） ” were significantly higher at 7 dpi. Expression of IL4 was significantly higher in LLLT group. Preservation of myelin sheath and BBB scores were significantly higher in LLLT group at 14dpi and 7,14dpi respectively. The present study indicates that low level laser therapy can promote the beneficial M2 macrophage/microglia polarization, reduce the demyelination and accelerate locomotor functional recovery after spinal cord injury in rats.

关 键 词：医用光学 弱激光治疗 脊髓损伤 巨噬细胞极化 

分 类 号：R318.51[医药卫生—生物医学工程]