中国科学院心理研究所机构知识库

目的 研究窄带蓝光经眼无创光疗对创伤后应激障碍(post-traumatic stress disorder,PTSD)模型大鼠的干预作用，以及光疗对基底外侧杏仁核(basolateral amygdala, BLA)区神经元激活水平的影响。方法 实验对象SD大鼠24只，随机分为空白对照组、应激对照组与光疗组三组，每组各8只大鼠。其中空白对照组大鼠正常饲养，不进行建模和光疗处理；应激对照组采用单次延长应激结合足底电击模型(single inescapable electric foot shock after single prolonged stress procedure, SPS-S)应激处理后，建立PTSD模型但不进行无创光疗干预；光疗组采用SPS-S法应激处理后建立PTSD模型，于应激处理后10 d开始，每天上午8～9点经眼进行波长470 nm窄带LED无创光疗干预，连续照射14 d，照度500 lux。分别于应激处理后第7天、无创光疗处理后第14天，利用旷场实验(open field test,OFT)、高架十字迷宫(elevated plus maze,EPM)检测大鼠的焦虑症状；使用木僵实验检测大鼠的恐惧症状，评估PTSD模型构建和效度、经眼无创光疗逆转应激诱导的焦虑行为情况。三组采用相应处理后，在行为学检测后1～1.5 h内，各取6只大鼠进行灌注和免疫荧光染色，观察c-fos表达水平以评估情绪调节重要脑区基底外侧杏仁核(basolateral amygdala, BLA)区的神经元激活水平。结果 (1)SPS-S法应激处理后第7天，旷场实验结果为应激对照组、光疗组大鼠进入中央区的次数、中央区停留时间均显著低于空白对照组(均P<0.001)；高架十字迷宫实验结果为应激对照组、光疗组大鼠进入开放臂的次数、开放臂停留时间比均显著低于空白对照组(均P<0.001)；木僵实验结果为应激对照组、光疗组大鼠与空白对照组比较，木僵时间百分比显著增加(均P<0.05)，成功构建PTSD动物模型。(2)经眼无创光疗处理后第14天，旷场实验结果为光疗组大鼠进入旷场中央区的次数、中央区停留时间均显著多于应激对照组(均P<0.05)，且与空白对照组比较差异无统计学意义(均P>0.05)；高架十字迷宫实验结果为光疗组大鼠进入开放臂次数与开放臂停留时间百分比均显著高于应激对照组(均P<0.001)，且显著低于空白对照组(均P<0.001)；木僵实验结果为光疗组的木僵时间百分比显著高于空白对照组(P<0.05)，与应激对照组比较，差异无统计学意义(均P>0.05)。免疫荧光结果为光疗组BLA的c-fos表达水平显著低于应激对照组(P<0.05)，而与空白对照组比较差异无统计学意义(P>0.05)。结论 经眼窄带蓝光LED光疗干预PTSD大鼠，可明显改善大鼠的焦虑样行为，降低BLA区的神经元激活水平。

Objective To study the intervention effect of non-invasive transocular phototherapy with narrow-band blue light on post-traumatic stress disorder（PTSD）model rats and the potential brain regions of phototherapy intervention. Methods Totally 24 SD rats were randomly allocated into a control group，stress group and light group. Single Inescapable Electric Foot Shock after Single Prolonged Stress Procedure（SPS-S for short）was used to construct PTSD model rats. 470 nm narrow-band blue light of the illuminance of 500 lux was used to perform non-invasive phototherapy intervention on SPS-S rats from 8：00 to 9：00 am each day in 2 weeks. The rats in the control group received neither SPS-S nor phototherapy. The rats in the stress group were given SPS-S but no phototherapy. The rats in the light group were given both SPS-S and phototherapy. Open field test and elevated plus maze test were used to determine the anxiety level of the rats. Freezing was used to evaluate their fear levels. 6 rats of the 3 groups sacrificed 1-1. 5 h after the freezing test and paraformaldehyde perfusion was applied for the preservation of brain samples. Immunofluorescence staining with c-fos was used to evaluate the changes in the activation level of neurons in the basolateral amygdala （BLA）. Results （1）Seven days after SPS-S：In the open field test，the number of entries（P<0. 001）and dwell time（P<0. 001）in the central area decreased more in the stress and phototherapy groups than in the control group. In elevated plus maze test，the number of entries（P<0. 001）and the percentage of dwell time（P<0. 001）in the open arm decreased more in the stress and phototherapy groups than in the control group. In freezing test，the percentage of freezing time（P<0. 05）increased in the stress and phototherapy groups. （2）Fourteen days after phototherapy：In the open field test，the number of entries（P<0. 05）and dwell time（P<0. 05）in the central area of the open field were much more in the phototherapy and control group than in the stress group. The results of elevated plus maze showed that the number of entries（P<0. 001）and the percentage of dwell time（P<0. 001）in the open arm increased significantly in the phototherapy group. The results of freezing test showed that there was no significant difference in the percentage of freezing time between the phototherapy group and stress group（P<0. 05），which were much higher than that in the control group（P>0. 05）. The immunofluorescence results indicated that the c-fos activation level of the light group was much lower than that of the stress group（P< 0. 05）and there was no significant difference between the light group and control group（P>0. 05）. Conclusions Transocular non-invasive phototherapy with narrow-band blue light（470±10）nm on SPS-S model will significantly ameliorate anxiety-like behaviors in rats and reduce the c-fos level of BLA.

海南省重大科技计划项目（ZDKJ2019012）;; 国家自然基金资助（61835009,82071517）;; 中国科学院心理研究所资助（E2CX4115CX）;; 中国医学科学院医学与健康科技创新工程（2019-I2M-5-061）