驻景丸加减方对形觉剥夺性近视小鼠线粒体动力学相关蛋白的影响

Effects of modified Zhujing Pill on mitochondrial dynamics related protein in mice with form deprivation myopia

摘要:

目的：探讨驻景丸加减方对形觉剥夺性近视小鼠视网膜色素上皮细胞线粒体结构及动力学相关蛋白的影响。方法：3周龄C57BL/6J小鼠随机分为对照组、模型组、中药组，各组均10只。模型组和中药组小鼠右眼采用形觉剥夺方法构建小鼠近视模型，形觉剥夺4wk后，中药组给予驻景丸加减方混悬液0.546g/(kg·d)(0.2mL/d)灌胃，共4wk，对照组、模型组给予等体积生理盐水灌胃。分别于实验开始时和实验结束时，应用A超检测小鼠右眼眼轴长度，带状光检影镜检测小鼠右眼屈光度。实验结束时，透射电镜观察视网膜色素上皮细胞线粒体超微结构，Western blot检测各组小鼠视网膜的线粒体融合蛋白1(MFN1)、视神经萎缩蛋白1(OPA1)、动力相关蛋白1(DRP1)表达，q-PCR检测MFN1、OPA1、DRP1基因表达。结果：实验前，对照组、模型组和中药组小鼠右眼屈光度和眼轴长度均无差异(P>0.05)。实验结束时，与对照组相比，模型组和中药组小鼠屈光度降低、伴眼轴持续延长(均P<0.05)，而中药组小鼠眼轴长度明显低于模型组，而屈光度高于模型组(均P<0.05)。Western blot和q-PCR结果示，与对照组相比，模型组和中药组MFN1、OPA1相对表达量均减少、DRP1相对表达量增加(均P<0.05)； 与模型组相比，中药组MFN1、OPA1相对表达量增加(均P<0.05)。电镜结果示，中药组可见小鼠右眼视网膜线粒体仅轻度肿胀，而模型组可见线粒体明显肿胀、嵴紊乱变空。结论：驻景丸加减方可能通过调控线粒体动力学关键蛋白MFN1、OPA1、DRP1发挥保护视网膜线粒体的作用，对轴性近视小鼠视网膜起保护作用。

Abstract:

AIM: To investigate the effects of modified Zhujing Pill on the mitochondrion structure and dynamin-related protein of retinal pigment epithelial cells(RPEs)in mice with form deprivation myopia.METHODS: 3-week-old C57BL/6J mice were randomly divided into control group, model group and Chinese medicine group, with 10 mice in each group. Myopia model of the right eye of mice was established by means of form deprivation in model and Chinese medicine groups. After 4wk, the Chinese medicine group were given intragastric administration of modified Zhujing Pill suspension 0.546g/(kg·d)(0.2mL/d)for 4wk, and same amount of saline was given to mice in other groups at the same time of modeling. The axial length and diopter of the right eye of the mouse were measured before and after the experiment by A-ultrasound and a strip retinoscope respectively. At the end of the experiment, the mitochondrial ultrastructure of RPEs was observed by transmission electron microscope. Western blot, and real-time fluorescent quantitative PCR(q-PCR)were used to detect quantitative and gene expression of mitofusin 1(MFN1), optic atrophy 1(OPA1), and dynamin-related protein 1(DRP1)in retinal tissues respectively.RESULTS: At the beginning of the experiment, there was no statistically significant difference in axial length and diopter of the right eye of the mouse in control, model and Chinese medicine groups(P>0.05). At the end of the experiment, compared with the control group, the mice in the model group and the Chinese medicine group had lower diopter and continuously prolonged axial length(all P<0.05), while the mice in the Chinese medicine group had significantly shorter axial length and higher diopter than the model group(all P<0.05). Western blot and q-PCR results showed that the relative expression of MFN1 and OPA1 decreased and DRP1 increased in both the model group and the Chinese medicine group compared with the control group(all P<0.05), and the relative expression of MFN1 and OPA1 increased in the Chinese medicine group compared with the model group(all P<0.05). The electron microscopic results showed that the mitochondria in the right retina of the mice were only mildly swollen in the Chinese medicine group, while the mitochondria in the model group were obviously swollen and disordered and empty.CONCLUSION: Modified Zhujing Pill could protect the retinal mitochondria by regulating the key proteins of mitochondrial dynamics(MFN1, OPA1, and DRP1), and it has a protective effect on the retina of axial myopic mice.