Evaluation of the Effectiveness of a Chronic Ocular Hypertension Mouse Model Induced by Intracameral Injection of Cross-Linking Hydrogel.
Evaluation of the Effectiveness of a Chronic Ocular Hypertension Mouse Model Induced by Intracameral Injection of Cross-Linking Hydrogel.
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前房注射交联水凝胶诱导慢性高眼压小鼠模型的有效性评价
DOI：
10.3389/fmed.2021.643402
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发表时间：
2021
影响因子：
3.9
通讯作者：
Zhong Y
中科院分区：
文献类型：
--
作者：
Chen J;Sun J;Yu H;Huang P;Zhong Y
背景：青光眼是一种不可逆的致盲性神经退行性疾病，以视网膜神经节细胞的进行性丧失为特征。目前青光眼动物模型不能提供慢性眼压升高，不能长时间维持视介质清晰度，给青光眼的研究带来一定困难。本研究建立了交联水凝胶眼内注射诱导小鼠慢性高眼压模型。方法：6 ~ 8周龄C57BL/6J小鼠随机分为对照组和手术组。手术组小鼠注射交联水凝胶诱导高眼压。术前、术后3天、每周测量眼压，直至研究结束。采用闪烁视觉诱发电位（F-VEP）观察慢性高眼压（COH）术后不同时间（术前及2、4、6周）视神经功能。在COH后2、4、6周，采用western blotting检测对照组和小鼠视网膜TNF-α、IL-1β和IL-17A蛋白的表达。免疫荧光染色和western blotting检测小胶质细胞活化情况。通过TUNEL实验和Brn3a蛋白标记，观察交联水凝胶在眼内注射2、4、6周后视网膜神经节细胞凋亡和丢失情况。采用神经丝重多肽蛋白标记法评价COH小鼠视神经轴突的损失情况。结果：眼内注射交联水凝胶可使眼内压（IOP）平均升高19.3±4.1 mmHg，且持续至少8周。COH小鼠与假手术小鼠的IOP差异有统计学意义（p < 0.0001）。成功率为75%。COH小鼠的F-VEP平均振幅较对照组降低（分别在COH后2、4、6周p = 0.0149、0.0012、0.0009），平均潜伏期较对照组延长（分别在COH后2、4、6周p = 0.0290、<0.0001、<0.0001）。COH小鼠TNF-α、IL-1β、IL-17A、Iba-1和CD68蛋白表达升高。在COH加工过程中，与对照组相比，小胶质细胞数量增加，细胞形态改变，体更圆，突起更粗。COH后2、4、6周小鼠视网膜神经节细胞（RGCs）凋亡明显（p = 0.0061、0.0012、<0.0001、0.0371，分别与对照组比较）。与对照组相比，COH小鼠的RGC密度在COH后2、4和6周分别显著降低（p = 0.0042、0.0036和<0.0001）。COH后2周、4周和6周，交联水凝胶注射后小鼠视神经轴突明显丧失（p = 0.0095、0.0002和<0.0001）。结论：单次眼内注射交联水凝胶可有效诱导小鼠慢性高眼压，导致视网膜神经节细胞进行性丧失，炎性细胞因子表达水平和小胶质细胞活化水平升高，视神经功能恶化。
Background: Glaucoma is an irreversible and blinding neurodegenerative disease that is characterized by progressive loss of retinal ganglion cells. The current animal models of glaucoma fail to provide a chronic elevated intraocular pressure and cannot maintain the optical media clarity for a long time, which brings some difficulties to the study of glaucoma. Here, we developed a new chronic ocular hypertension model of mice induced by cross-linking hydrogel intracameral injection. Methods: C57BL/6J mice aged 6–8 weeks were randomly divided into the control group and the operation group. The mice of the operation group were injected with cross-linking hydrogel to induce ocular hypertension. Intraocular pressure was measured preoperatively, 3 days after surgery, and weekly until the end of the study. Flash visual evoked potential (F-VEP) was used to observe optic nerve function at different times (preoperatively and 2, 4, and 6 weeks) after chronic ocular hypertension (COH). Retinal TNF-α, IL-1β, and IL-17A protein expression were measured by western blotting in the control group and in mice at 2, 4, and 6 weeks after COH. Microglial cell activation was evaluated by immunofluorescence staining and western blotting. Apoptosis and loss of retinal ganglion cells after 2, 4, and 6 weeks of intracameral injection of cross-linking hydrogel were observed by the TUNEL assay and Brn3a protein labeling. The loss of optic nerve axons in COH mice was evaluated by neurofilament heavy polypeptide protein labeling. Results: Intracameral injection of the cross-linking hydrogel induces increased intraocular pressure (IOP) to a mean value of 19.3 ± 4.1 mmHg, which was sustained for at least 8 weeks. A significant difference in IOP was noted between COH mice and sham-operation mice (p < 0.0001). The success rate was 75%. The average amplitude of F-VEP in mice with COH was reduced (p = 0.0149, 0.0012, and 0.0009 at 2, 4, and 6 weeks after COH vs. the control group, respectively), and the average latent period in mice with COH was longer (p = 0.0290, <0.0001, and <0.0001 at 2, 4, and 6 weeks after COH vs. the control group, respectively) compared with that in the control group. TNF-α, IL-1β, IL-17A, Iba-1, and CD68 protein expression increased in COH mice. During the processing of COH, the number of microglial cells increased along with cellular morphological changes of rounder bodies and thicker processes compared with the control group. Apoptosis of retinal ganglion cells (RGCs) was clearly observed in mice at 2, 4, and 6 weeks after COH (p = 0.0061, 0.0012, <0.0001, and 0.0371 at 2, 4, and 6 weeks after COH vs. the control group, respectively). The RGC density decreased significantly in the COH mice compared with the control group (p = 0.0042, 0.0036, and <0.0001 at 2, 4, and 6 weeks after COH vs. the control group, respectively). There was a significant loss of optic nerve axons in mice after intracameral injection of cross-linking hydrogel (p = 0.0095, 0.0002, and <0.0001 at 2, 4, and 6 weeks after COH vs. the control group, respectively). Conclusions: A single intracameral injection of cross-linking hydrogel can effectively induce chronic ocular hypertension in mice, which causes progressive loss of retinal ganglion cells, increased expression levels of inflammatory cytokines and microglial cell activation, and deterioration of optic nerve function.
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DOI：
10.1111/1440-1681.13141
发表时间：
2019-10-01
影响因子：
2.9
作者：
Ren, Yongbo;Qi, Yanxiu;Su, Xingjie
通讯作者：
Su, Xingjie
影响因子：
4.4
作者：
Liu, Hsin-Hua;Flanagan, John G.
通讯作者：
Flanagan, John G.
影响因子：
9.3
作者：
Cherry JD;Olschowka JA;O'Banion MK
通讯作者：
O'Banion MK
影响因子：
1.2
作者：
Feng, Liang;Chen, Hui;Liu, Xiaorong
通讯作者：
Liu, Xiaorong
影响因子：
3.4
作者：
Moreno, MC;Marcos, HJA;Rosenstein, RE
通讯作者：
Rosenstein, RE