(1)背景:眼压(IOP)升高和视网膜神经节细胞减少是小鼠慢性高眼压有效建模的常见指标。在这项研究中,评估了小鼠模型对药物治疗降低眼压的敏感性,使用细胞毒性试验证实了模型的安全性,通过评估眼压和神经节细胞层神经元的改变来评估高眼压小鼠模型的成功率。(2)方法:本研究采用光交联丝胶凝胶注射和LED灯照射制备慢性高眼压小鼠模型。25只C57BL/6雄性小鼠的眼睛在左眼前房中注射5μL丝胶凝胶后,从正面接受405nm的紫外光照射2分钟。每天测量小鼠的眼压,眼压升高超过5mmHg被认为是高眼压。当眼压降低时,重复了一次干预,但治疗之间的间隔至少为2周。右眼不作为正常对照组进行任何治疗。小鼠眼球被HE染色,Ni型,和免疫荧光来评估模型的功效。眼压稳定四周后,提供两种常见药物(他氟前列素滴眼液和噻吗洛尔滴眼液)一周。和IOP变化进行评估以确定慢性高眼压小鼠模型的药物敏感性。此外,利用CellTiter96®AQuequity单溶液细胞增殖测定(MTS)通过评估可光交联丝胶凝胶对细胞的有害作用来研究高眼压模型的安全性。(3)结果:注射前,实验组基础眼压为(9.42±1.28)mmHg(1kPa=7.5mmHg),对照组为(9.08±1.21)。注射后,白内障发生在一只眼睛,一只眼睛角膜水肿,一只眼睛的眼内炎,一只眼睛的虹膜监禁,一只眼睛和眼球萎缩。五只有并发症的小鼠被排除在实验之外,剩下二十只老鼠。注射后四周,实验组眼压维持在(19.7±4.52)mmHg,对照组维持在(9.92±1.55)mmHg,两组间差异有统计学意义(p<0.05)。在干预之前,高眼压对照组的眼压为(21.7±3.31)mmHg,他氟前列素滴眼液组(20.33±2.00)mmHg,马来酸噻吗洛尔滴眼液组(20.67±3.12)mmHg。干预后眼压为(23.2±1.03)mmHg,(12.7±2.11)mmHg,和(10.4±1.43)mmHg,分别。干预前后,高眼压对照组差异无统计学意义(p>0.05),噻吗洛尔滴眼液组差异有统计学意义(p<0.05),与他氟前列素滴眼液组比较,差异有统计学意义(p<0.05)。停药一周后,三组间眼压差异无统计学意义(p>0.05)。在高IOP组,蛋白质(丝胶凝胶)在前房中显示出短条或碎片结构,伴有大量的巨噬细胞和少量的浆细胞。空白对照组室角形状正常。前房注射丝胶凝胶8周后,视网膜神经节细胞数量明显减少,与空白对照组比较差异有统计学意义(p<0.05)。用可光交联的丝胶水凝胶处理细胞后,与空白对照组相比,MTS的CellTiter96®测定试剂盒的数据没有显着差异(p>0.05)。(4)结论:前房注射丝胶并紫外光照射可成功建立小鼠慢性高眼压模型。该模型可以模拟青光眼的结构和功能变化,在大多数降压药物作用后可以有效降低IOP,它对药物高度敏感。丝胶对细胞无明显毒性作用,安全性高。
(1) Background: A rise in intraocular pressure (IOP) and decreased
retinal ganglion cells are frequent indicators of effective modeling of chronic ocular hypertension in mice. In this study, the sensitivity of the mouse model to pharmaceutical therapy to reduce intraocular tension was assessed, the model\'s safety was confirmed using a cytotoxicity test, and the success rate of the mouse model of ocular hypertension was assessed by assessing alterations in IOP and neurons in the ganglion cell layer. (2) Methods: A mouse model of chronic ocular hypertension was produced in this study by employing photocrosslinkable sericin hydrogel injection and LED lamp irradiation. The eyes of 25 C57BL/6 male mice were subjected to 405 nm UV light from the front for 2 min after being injected with 5 μL of sericin hydrogel in the anterior chamber of the left eye. IOP in the mice was measured daily, and IOP rises greater than 5 mmHg were considered intraocular hypertension. When the IOP was lowered, the intervention was repeated once, but the interval between treatments was at least 2 weeks. The right eyes were not treated with anything as a normal control group. Mice eyeballs were stained with HE, Ni-type, and immunofluorescence to assess the model\'s efficacy. Two common drugs (tafluprost eye drops and timolol eye drops) were provided for one week after four weeks of stable IOP, and IOP changes were assessed to determine the drug sensitivity of the mouse model of chronic ocular hypertension. Furthermore, CellTiter 96® AQueous One Solution Cell Proliferation Assay (MTS) was utilized to investigate the safety of the ocular hypertension model by evaluating the deleterious effects of photocrosslinkable sericin hydrogel on cells. (3) Results: Before injection, the basal IOP was (9.42 ± 1.28) mmHg (1 kPa = 7.5 mmHg) in the experimental group and (9.08 ± 1.21) in the control group. After injection,
cataract occurred in one
eye, corneal edema in one
eye, endophthalmitis in one
eye, iris incarceration in one
eye, and eyeball atrophy in one
eye. Five mice with complications were excluded from the experiment, and twenty mice were left. Four weeks after injection, the IOP of the experimental group was maintained at (19.7 ± 4.52) mmHg, and that of the control group was maintained at (9.92 ± 1.55) mmHg, and the difference between the two groups was statistically significant (p < 0.05). Before the intervention, the IOP in the experimental group was (21.7 ± 3.31) mmHg in the high IOP control group, (20.33 ± 2.00) mmHg in the tafluprost eye drops group, and (20.67 ± 3.12) mmHg in the timolol maleate eye drops group. The IOP after the intervention was (23.2 ± 1.03) mmHg, (12.7 ± 2.11) mmHg, and (10.4 ± 1.43) mmHg, respectively. Before and after the intervention, there were no significant differences in the high-IOP control group (p > 0.05), there were statistically significant differences in the timolol eye drops group (p < 0.05), and there were statistically significant differences in the tafluprost eye drops group (p < 0.05). One week after drug withdrawal, there was no significant difference in IOP among the three groups (p > 0.05). In the high-IOP group, the protein (sericin hydrogel) showed a short strips or fragmented structure in the anterior chamber, accompanied by a large number of macrophages and a small number of plasma cells. The shape of the chamber angle was normal in the blank control group. The number of
retinal ganglion cells decreased significantly 8 weeks after injection of sericin hydrogel into the anterior chamber, and the difference was statistically significant compared with the blank control group (p < 0.05). After the cells were treated with photocrosslinkable sericin hydrogel, there was no significant difference in the data of the CellTiter 96® assay kit of MTS compared with the blank control group (p > 0.05). (4) Conclusions: A mouse model of chronic intraocular hypertension can be established successfully by injecting sericin in the anterior chamber and irradiating with ultraviolet light. The model can simulate the structural and functional changes of
glaucoma and can effectively reduce IOP after the action of most antihypertensive drugs, and it is highly sensitive to drugs. Sericin has no obvious toxic effect on cells and has high safety.