[1]巩一博,郭雅图,张伟.菊花提取液对小鼠视网膜光损伤的保护作用[J].眼科新进展,2021,41(2):110-115.[doi:10.13389/j.cnki.rao.2021.0023]
　GONG Yibo,GUO Yatu,ZHANG Wei.Protective effect of chrysanthemum extract against retinal light damage in mice[J].Recent Advances in Ophthalmology,2021,41(2):110-115.[doi:10.13389/j.cnki.rao.2021.0023]
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《眼科新进展》[ISSN:1003-5141/CN:41-1105/R]

卷:
41卷
期数:
2021年2期
页码:
110-115
栏目:
实验研究
出版日期:
2021-02-05

文章信息/Info

Title:
Protective effect of chrysanthemum extract against retinal light damage in mice
作者:
巩一博; 郭雅图; 张伟
300020　天津市，天津医科大学眼科临床学院（巩一博，张伟）；300020　天津市，天津市眼科医院（郭雅图，张伟）
Author(s):
GONG Yibo1; GUO Yatu2; ZHANG Wei1; 2
1.Clinical College of Ophthalmology,Tianjin Medical University，Tianjin 300020，China
2.Tianjin Eye Hospital，Tianjin 300020，China
关键词:
光损伤; 老年性黄斑变性; 菊花; 抗氧化作用; 凋亡
Keywords:
light damage;  age-related macular degeneration;  chrysanthemum;  antioxidation;  apoptosis
分类号:
R774
DOI:
10.13389/j.cnki.rao.2021.0023
文献标志码:
A
摘要:
目的　研究菊花提取液对小鼠视网膜光损伤是否具有保护作用。方法　取28只C57BL/6J小鼠随机分为对照组（8只）、模型组（8只）、菊花低剂量组（6只）及菊花高剂量组（6只）。对照组及模型组普通饲料喂养，菊花低剂量及高剂量小鼠分别给予0.23 g·kg-1及0.38 g·kg-1菊花提取液连续灌胃8周，12 h明暗交替饲养，对照组不做处理，其余三组制作光损伤模型。评价视网膜电生理（ERG）功能；光学相干断层扫描（OCT）观察小鼠活体状态下视网膜形态，并通过HE染色与离体状态下视网膜形态作对比；视网膜荧光素眼底血管造影（FFA）获取活体状态下小血管血流图像，并通过Angiotool分析动静脉期小血管面积、小血管面积百分比和血管分支点个数；Tunel免疫荧光染色检测视网膜细胞凋亡状态，Image J分析并比较各组视网膜神经节细胞（RGC）层、内核层(INL)、外核层(ONL)及总体细胞凋亡率；取眼动脉血行超氧化物歧化酶和过氧化氢酸活性检测。结果　OCT检测结果显示，模型组视网膜色素上皮（RPE）层出现弓背向上的高反射信号，而对照组、菊花低剂量及菊花高剂量组未出现明显视网膜异常改变。HE 染色结果显示，模型组RPE层、光感受器细胞层结构破坏，而对照组、菊花低剂量组及菊花高剂量组小鼠视网膜相对完整，未出现明显结构异常。ERG检查结果显示，0.01 cds·m-2暗适应下b波振幅：模型组较对照组降低41.63%，菊花低剂量组较模型组提高83.50%，菊花高剂量组较模型组提高120.60%；3.00 cds·m-2暗适应下a波振幅：模型组较对照组降低22.28%，菊花低剂量组较模型组提高76.45%，菊花高剂量组较模型组提高118.50%；3.00 cds· m-2明适应条件下a波振幅：模型组较对照组降低37.04%，而菊花高剂量组较模型组提高76.45%;以上差异均有统计学意义（均为P<0.05）。FFA检查结果显示，模型组的小血管面积、小血管面积百分比、血管分支点个数均较对照组提高，差异均有统计学意义（均为P<0.05）。RGC 细胞凋亡结果显示，菊花低剂量组较模型组凋亡率降低73.18%，菊花高剂量组较模型组降低57.28%；INL细胞凋亡结果显示，菊花低剂量组较模型组凋亡率降低78.31%，菊花高剂量组较模型组降低52.75%；以上差异均有统计学意义（均为P<0.01）。超氧化物歧化酶在各组中无明显差异，而菊花低剂量组及菊花高剂量组过氧化氢酶与模型组相比差异均有统计学意义（均为P<0.05）。结论　光损伤造成了小鼠视网膜形态及ERG功能的异常，出现了类似于老年性黄斑变性样视网膜改变，而预先给予一定浓度的菊花提取液对光损伤造成的视网膜病变有一定的保护性作用。
Abstract:
Objective　To study the protective effect of chrysanthemum extract on retinal light injury in mice.Methods　28 C57BL/6J mice were randomly divided into control group,light damaged group,low-dose chrysanthemum group and high-dose chrysanthemum group.The mice in the control and the light damaged model group were fed with normal diet,and low-dose and high-dose chrysanthemum mice were given continuously intragastric administration of chrysanthemum extract 0.23 g·kg-1 and 0.38 g·kg-1 for 8 weeks,12 hours daylight and dark alternately. except for the control group,the other three groups were exposed to light to establish light damaged model.The electrophysiological function of retina was evaluated by electroretinogram (ERG); the morphology of retina in vivo was observed by optical coherence tomography (OCT),and compared with that in vitro by HE staining.Results　The results of OCT detection showed that the retinal pigment epithelial (RPE) cells in the light damaged group showed a signal of high reflection and arch formed,while there was no obvious retinal abnormality in the control group,low-dose chrysanthemum and high-dose chrysanthemum group.Damaged structure of the photoreceptor cell layer and RPE were showed in the HE staining outcomes.While in the control group,low-dose chrysanthemum and high-dose chrysanthemum group,the retina was relatively intact and there was no obvious structural abnormality.The results of retinal ERG showed that in the comparison of b waves under 0.01 cds·m-2 dark adaptation,the amplitude of light damaged group was 41.63% lower than that of control group,low-dose chrysanthemum group was 83.50% higher than that of light damaged group and high-dose chrysanthemum group was 120.60% higher.In the comparison of a wave under 3.00 cds ·m-2 dark adaptation,the light damaged group decreased by 22.28%,the low dose chrysanthemum group increased by 76.45% compared with the light damaged group,and the high-dose chrysanthemum group increased by 118.50%.The above differences were statistically significant (all P<0.05).In the a-wave comparison under the condition of 3.00 cds·m-2 adaptation,the light damaged group decreased by 37.04% compared with the control group,while the high-dose chrysanthemum group increased by 76.45% compared with the light damaged group,all the differences were statistically significant (all P<0.05).The FFA results showed that the area of small vessels the percentage of small vessels and the number of vascular branches was higher than that in the control group,all the differences were statistically significant (all P<0.05).The apoptosis rate of RGC cells in the low-dose chrysanthemum group was 73.18% lower than that in damaged group,and in the high-dose chrysanthemum group was 57.28% lower than that in the light damaged group.The results of INL cell apoptosis showed that the apoptosis rate in the low-dose chrysanthemum group decreased by 78.31%,and that in the high dose chrysanthemum group decreased by 52.75% compared with the light damaged group.The above differences were statistically significant (all P<0.01).The serum biochemical tests of superoxide dismutase (SOD) showed that there was no significant difference among each group,but the catalase in the low-dose and high-dose of chrysanthemum groups were higher than that in the light damaged group with difference of statistically significant (all P<0.05).Conclusion　Light damage causes abnormal retinal morphology and ERG in mice,which is similar to age-related macular degeneration changes.However pre-administration of a certain concentration of chrysanthemum extract has a certain protective effect on retinopathy caused by light damage.

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备注/Memo

备注/Memo:
国家自然科学基金青年基金 (编号：81300791)；天津市自然科学基金（编号：18JCYBJC26500）；天津市重点研发计划科技支撑重点项目（编号：S19ZC63957）
更新日期/Last Update: 2021-02-05