方法：C57BL/6小鼠30只随机分为阴性对照组、近视模型组以及中药干预组，每组10只。除了阴性对照组外，近视模型组、中药干预组小鼠均使用半透明EP管遮盖右眼制成形觉剥夺性近视(FDM)模型； 中药干预组灌胃驻景丸加减方混悬液0.546g/(kg·d)(0.15mL/d)，阴性对照组、近视模型组灌胃等量生理盐水(0.15mL/d)，共4wk。分别于实验开始、实验结束，使用带状检影镜测量小鼠右眼屈光度，A超测量小鼠右眼眼轴长度。实验结束时，取所有小鼠右眼进行检测，免疫荧光法定位和检测视网膜小胶质细胞标志物(Iba1)活性与迁移； 透射电镜观察视网膜色素上皮细胞中自噬小体形成情况； Western Blot、实时荧光定量PCR(q-PCR)检测视网膜组织自噬标志物LC3Ⅱ和p62蛋白定量及基因表达情况。

结果：实验结束时小鼠右眼屈光度示，近视模型组、中药干预组形成相对近视，近视模型组、中药干预组较阴性对照组显著降低(均P<0.01)。实验结束时，近视模型组、中药干预组眼轴长度较阴性对照组眼轴长度显著增加(P<0.01)。免疫荧光法定位和检测Iba1示，近视模型组视网膜中Iba1的平均光密度增加趋势最明显，阴性对照组增高趋势次之，中药干预组有降低趋势，近视模型组较阴性对照组显著增加(P<0.05)，中药干预组较近视模型组显著降低(P<0.05)，且发现近视模型组、中药干预组Iba1向神经节细胞层迁移。透射电镜示，近视模型组、中药干预组视网膜色素上皮细胞中观察到自噬小体。Western Blot法、q-PCR检测结果示，LC3Ⅱ、p62表达在中药干预组增加趋势最明显、近视模型组其次、阴性对照组最低。

结论：驻景丸加减方可能通过抑制小胶质细胞活化，增强FDM小鼠视网膜自噬。

METHODS: Thirty C57BL/6 mice were randomly divided into a negative control group, a myopia model group and a traditional Chinese medicine intervention group, with 10 mice in each group. Except for the negative control group, all mice in the myopia model group and the traditional Chinese medicine intervention group used translucent EP tubes to cover their right eyes to make a form deprivation myopia(FDM)model; The traditional Chinese medicine intervention group gavage Zhujing pill modified suspension 0.546g/(kg·d)(0.15mL/d), the negative control group and the myopia model group were given an equal amount of normal saline(0.15mL/d)for 4wk. At the beginning and the end of the experiment respectively, the right eye diopter of the mouse was measured with a strip retinoscope, measurement of the axial length of the right eye of mouse by A-ultrasound. At the end of the experiment, the right eyes of all mice were taken for detection, and immunofluorescence method was used to locate and detect the activity and migration of the retinal microglia marker(Iba1); Transmission electron microscope observation of autophagosome formation in retinal pigment epithelial cells; Western Blot, real-time fluorescent quantitative PCR(q-PCR)to detect the autophagy marker LC3Ⅱ and p62 protein quantitative and gene expression in retinal tissues.

RESULTS: At the end of the experiment, the refractive power of the right eyes of mice showed that the myopia model group and the traditional Chinese medicine intervention group formed relative myopia, the myopia model group and the traditional Chinese medicine intervention group were significantly lower than those of the negative control group(all P<0.01). At the end of the experiment, the axial length of the myopia model group and the Chinese medicine intervention group were significantly increased compared with the negative control group(P<0.01). Immunofluorescence method for locating and detecting Iba1 showed that the average optical density of Iba1 in the retina of the myopia model group increased the most obviously, followed by the increase in the negative control group, and the decrease in the traditional Chinese medicine intervention group. Compared with the negative control group, the myopia model group increased significantly(P<0.05), and the traditional Chinese medicine intervention group was significantly lower than the myopia model group(P<0.05). It was found that Iba1 migrated to the ganglion cell layer in the myopia model group and the traditional Chinese medicine intervention group. Transmission electron microscopy showed that autophagosomes were observed in the retinal pigment epithelial cells of the myopia model group and the Chinese medicine intervention group. The results of Western Blot and q-PCR showed that the expression of LC3Ⅱ and p62 increased most obviously in the traditional Chinese medicine intervention group, followed by the myopia model group, and the negative control group was the lowest.

CONCLUSION: The results of the study show that modified Zhujing pill may enhance retinal autophagy in mice with FDM by inhibiting the activation of microglia.