Effective improvement of retinal pathological injury and autophagy changes induced by explosive eye blast injury by protective goggles
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摘要: 首先基于头部动态测试系统与激波管和外场实弹实爆测试环境，验证了眼部装备（护目镜和风镜）的防护性能。研究结果表明，风镜防护性能更优，建议给执勤人员配发兼容防紫外、强光、烟雾和防破片功能的风镜产品，以提高相关人员眼部防护能力。此后，研究眼部爆震伤后组织损伤、功能改变及市售风镜动物实验版的防护作用与机制，为防治眼部爆震伤及风镜后续的设计改进提供理论依据。选用比格犬和C57小鼠进行相关动物实验，通过HE、Tunel、Nissl染色、视觉电生理检查等检测方法，发现随着冲击波强度的提高和伤后时间的延长，视网膜损伤程度和细胞凋亡程度均提高，其中神经节细胞层（ganglion cell layer, GCL）和光感受器内外节交界处（layer of photoreceptor inner/outer segments, IS/OS）受到的损伤最严重；进一步研究分子变化发现，自噬相关调节蛋白SQSTM1/p62（P <
0.0001 ）和LC3-Ⅱ（P =0.8437 )、LC3-Ⅰ（P = 0.003）的表达量明显增高，说明视网膜损伤一定程度上是由爆震伤后自噬增强这一机制导致的。市售风镜的动物实验版能够有效减轻冲击波对视网膜的损伤，保护RNFL、INL/ONL、GCL和IS/OS的结构。同时，与其他组相比，3.5 MPa组防护组与损伤组视网膜损伤和细胞凋亡程度差异最显著，推测该强度下防护风镜发挥了最大的保护作用，保护机制与防护降低视网膜细胞自噬相关。Abstract: We firstly verified the protective performance of eye equipment (goggles) based on a head dynamic test system and shock tube and field live blast test environments. The results show that goggles have better protective performance and suggest that duty personnel should be equipped with goggles that have combined functions of anti-ultraviolet, anti-glare, anti-smoke and anti-fragmentation in to improve eye protection capabilities. After that, we investigated the tissue damage and functional impairment changes after explosive eye blast injury, and the protective effect and mechanism of the goggles for animal experimental version. This may provide a theoretical basis for prevention and treatment, and also have important implications for the design and improvement of protective goggles. Beagles and C57 mice were used for related animal experiments, and the changes in retinal layer thickness and cell apoptosis were observed after blast injury by HE, Tunel, Nissl staining, visual electrophysiology detection and other methods. It was found that with the increase of blast intensity and the extension of time after explosion, both the degree of retinal injury and cell apoptosis increased, among which the ganglion cell layer and photoreceptor inner and outer segments suffered the most severe damage. Further research on molecular changes indicates that the expression levels of autophagy-related regulatory proteins SQSTM1/p62 (P <0.0001 ) and LC3-Ⅱ (P =0.8437 ), as well as LC3-Ⅰ (P = 0.003), are significantly increased, suggesting that retinal damage is, to some extent, induced by autophagic mechanisms. The protective goggles could effectively reduce the damage of blast wave to retina, protect the structural integrity of retinal nerve fiber layer, inner and outer nuclear layer, ganglion cell layer and photoreceptor inner and outer segments. At the same time, compared with that of other groups, the difference in retinal layer thickness and cell apoptosis was most significant in the 3.5 MPa group, suggesting that the glasses played the maximum protective effect at this intensity, which may be related to the reduction in the retinal autophagy.-
Key words:
- beagles /
- explosive eye blast injury /
- protective goggles /
- retina /
- mitochondria /
- autophagy
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图 2 太阳镜和风镜的防护性能测试实验装置
Figure 2. Experimental devices for testing protective performances tof sun glasses and goggles
图 3 3种不同防护条件下的激波管管口和头模眼部超压-时间曲线
Figure 3. Overpressure-time curves of the mouth of the shock tube and the eyes of the head models under three different protection conditions
图 4 3种不同防护条件下的自由场和头模眼部超压-时间曲线
Figure 4. Overpressure-time curves of the free field and the eyes of the head models under three different protection conditions
图 5 不同激波强度下防护组与损伤组视网膜HE染色结果
Figure 5. Retinal HE staining results of the protection group and the injury group under different shock wave intensities
图 6 不同激波强度下样本平均INL和ONL厚度
Figure 6. Mean INL and ONL thickness of the samples under different shock wave intensities
图 7 4.5 MPa激波下，不同取样时间防护组与损伤组视网膜HE染色结果
Figure 7. Retinal HE staining results of the protection group and the injury group under 4.5-MPa shock wave at different sampling times
图 8 4.5 MPa激波下，不同取样时间样本平均INL和ONL厚度以及两者的比
Figure 8. Mean INL and ONL thicknesses of the samples and their ratio under 4.5-MPa shock wave at different sampling times
图 9 不同激波强度下防护组与损伤组视网膜Tunel染色结果
Figure 9. Retinal Tunel staining results of the protection group and the injury group under different shock wave intensities
图 10 4.5 MPa激波下，不同取样时间防护组与损伤组视网膜Tunel染色结果
Figure 10. Retinal Tunel staining results of the protection group and the injury group under 4.5-MPa shock wave at different sampling times
图 11 不同激波强度下样本IS/OS厚度和
Figure 11. Sum of IS and OS thickness of the samples under different shock wave intensities
图 12 4.5 MPa激波下，不同取样时间样本IS/OS厚度和
Figure 12. Sum of IS and OS thickness of the samples under 4.5-MPa shock wave at different sampling times
图 13 不同激波强度下防护组与损伤组RGC细胞数
Figure 13. RGC count of the protection group and the injury group under different shock wave intensities
图 14 4.5 MPa激波下，不同取样时间防护组与损伤组RGC细胞数
Figure 14. RGC count of the protection group and the injury group under 4.5-MPa shock wave at different sampling times
图 15 不同激波强度下，防护组与损伤组单位视野下RGC凋亡数量
Figure 15. Apoptotic number of RGCs in unit field of the protection group and the injury group under different shock wave intensities
图 16 4.5 MPa激波下，不同取样时间防护组与损伤组单位视野下RGC凋亡数量
Figure 16. Apoptotic number of RGCs in unit field of the protection group and the injury group under 4.5-MPa shock wave at different sampling times
图 17 不同激波强度下，防护组与损伤组单位视野下凋亡细胞数量
Figure 17. Apoptotic number of cells in unit field of the protection group and the injury group under different shock wave intensities
图 18 不同激波强度下防护组与损伤组视网膜Nissl染色结果
Figure 18. Retinal Nissl staining results of the protection group and the injury group under different shock wave intensities
图 19 4.5 MPa激波下，不同取样时间防护组与损伤组视网膜Nissl染色结果
Figure 19. Retinal Nissl staining results of the protection group and the injury group under 4.5-MPa shock wave at different sampling times
图 20 不同激波强度下，防护组与损伤组单位视野下阳性细胞数
Figure 20. Number of positive cells in unit field of the protection group and the injury group under different shock wave intensities
图 21 4.5 MPa激波下，不同取样时间防护组与损伤组单位视野下阳性细胞数
Figure 21. Number of positive cells in unit field of the protection group and the injury group under 4.5-MPa shock wave at different sampling times
图 22 4.5 MPa激波下，防护组与损伤组视网膜HE和Nissl染色结果
Figure 22. Retinal HE staining and Nissl staining results of the protection group and the injury group under 4.5-MPa shock wave intensities
图 23 4.5 MPa激波下，样本平均INL和ONL厚度
Figure 23. Mean INL and ONL thicknesses of the samples under 4.5-MPa shock wave intensities
图 24 4.5 MPa激波下，防护组与损伤组单位视野下RGC细胞数
Figure 24. RGC and positive cell count in unit field of the protection group and the injury group under 4.5-MPa shock wave intensity
图 25 4.5 MPa激波下，防护组与损伤组视网膜Tunel检测结果
Figure 25. Retinal Tunel test results of the protection group and the injury group under 4.5-MPa shock wave intensity
图 26 防护组和损伤组视网膜的HE染色和Nissl染色结果以及各组视网膜厚度量化结果
Figure 26. Retinal HE and Nissl staining results of the protection group and the injury group as well as quantitative results of retinal thickness in each group
图 27 防护组和损伤组视网膜的凋亡检测以及视网膜GCL凋亡细胞量化结果
Figure 27. Apoptosis detection in the retina of the protection group and the injury group as well as quantitative results of apoptotic cells in the retinal RGC layer
图 28 视觉电生理检查各组Ops数据以及Ops指标中O2幅值量化
Figure 28. Ops data of each group measured by visual electrophysiology as well as quantification of O2 amplitude in Ops indices
图 29 视觉电生理检查各组VEP数据以及VEP指标中潜伏期量化和P2幅值量化
Figure 29. VEP data of each group measured by visual electrophysiology as well as quantifications of latency and P2 amplitude in VEP indices
图 30 透射电镜检测各组RGC中自噬情况
Figure 30. Transmission electron microscopy detection of autophagy in the retina of each group
图 31 WB检测各组中自噬相关蛋白SQSTM1/p62、LC3-Ⅰ和LC3-Ⅱ表达水平
Figure 31. WB detection of autophagy-related proteins SQSTM1/p62, LC3-Ⅰ and LC3-Ⅱ expression levels in each group
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