湿性年龄相关性黄斑变性脾虚证大鼠模型的建立与评价
Establishment and Evaluation of a Rat Model Integrating Spleen Deficiency Syn-drome and wet Age-related Macular Degeneration Pathology
李晓宇 1梁丽娜 1李佳豪 2高云 1陈强2
作者信息
- 1. 中国中医科学院眼科医院,北京 100040;中国中医药循证医学中心眼科疾病项目组,北京 100040
- 2. 中国中医科学院眼科医院,北京 100040
- 折叠
摘要
目的 使用大黄水煎液灌胃联合视网膜光凝建立湿性年龄相关性黄斑变性(wAMD)脾虚证大鼠模型,并对模型进行评价.方法 将24只10周龄BN大鼠随机分为对照组(CG)、光凝组(PG)、大黄联合光凝组(RPG),每组8只.RPG组大鼠予10 mL/kg灌胃大黄水煎液,行右眼视网膜氪激光光凝诱导脉络膜新生血管(CNV);PG组仅予氪激光光凝和等剂量蒸馏水灌胃;CG组仅灌胃蒸馏水,均连续干预10 d.记录3组大鼠10 d内的一般情况及表观行为情况;光凝后7 d、14 d活体进行眼底照相、荧光素眼底血管造影(FFA)、光学相干断层扫描(OCT)观察CNV病灶.14 d后取材,计算大鼠脾脏及胸腺指数,间苯三酚法检测血清D-木糖表达,测定血清α-淀粉酶活力;分离大鼠眼球视网膜色素上皮(RPE)层—脉络膜—巩膜的复合体,AF488偶联同工凝集素B4法观察CNV病灶;苏木精—伊红(HE)染色观察视网膜形态.结果 (1)一般情况及表观行为:RPG组大鼠毛发色泽、四肢肌肉、行动、精神状态等得分持续下降,体质量、体温低于PG组,差异均有统计学意义(t体质量=6.014,P= 0.000;t体温=3.125,P=0.005).(2)脾脏及胸腺指数:3组间大鼠胸腺及脾脏指数比较,差异均无统计学意义(P＞0.05).(3)血清D-木糖与α-淀粉酶:PG、RPG组大鼠血清D-木糖水平均低于CG组(tPG=11.270,tRPG=17.990,均P=0.000),且RPG组低于PG组(t=6.720,P=0.000),差异均有统计学意义;而 3 组间血清α-淀粉酶差异无统计学意义(P＞0.05).(4)眼底CNV:RPG组大鼠第7天、第14天CNV渗出面积高于PG组(t7 d=2.217,t14 d=2.210,均P=0.032),且灰度值高于PG组(t7 d=3.448,P=0.001;t14 d=2.215,P=0.032),差异均有统计学意义.(5)视网膜形态结构:CG组大鼠视网膜结构完整清晰,PG、RPG组视网膜结构破坏,伴有出血、水肿以及新生血管生成,且RPG组较PG组更为显著.(6)视网膜病变面积:RPG组平均病灶面积高于PG组,差异有统计学意义(t=4.645,P=0.000).结论 大黄水煎液灌胃联合视网膜激光光凝法可以成功建立脾虚证wAMD病证结合大鼠模型.
Abstract
OBJECTIVE To establish and evaluate a rat model of spleen deficiency syndrome combined with wet age-related macular degeneration(wAMD)using intragastric administration of rhubarb decoction combined with retinal photocoagulation.METHODS A total of 24 10-week-old BN rats were randomly divided into control group(CG),photocoagulation group(PG),and rhubarb combined with photocoagulation group(RPG),with eight rats in each group.Rats in RPG group were intragastrically administered rhubarb decoction twice daily at a dose of 10 mL/kg and underwent retinal krypton laser photocoagulation to induce choroidal neovascularization(CNV)in the right eye.The PG group received krypton laser photocoagulation and intragastric administration of an equivalent dose of distilled water.The CG group received intragastric administration of distilled water only.The interventions were administered continuously for 10 days.The general condition and behavioral changes of the rats were recorded.Fundus photography,fundus fluorescein angiography(FFA),and optical coherence tomography(OCT)were performed on live rats at seven and 14 days after photocoagulation to observe CNV lesions.After 14 days,the rats were sacrificed,and the spleen and thymus indices were calculated.Serum D-xylose expression was detected using the resorcinol method,and serum α-amylase activity was measured.The complex of the retinal pigment epithelium(RPE)layer,choroid,and sclera was isolated from the rat eyeballs,and CNV lesions were observed using AF488-conjugated isolectin B4.Hematoxylin and eosin(HE)staining was performed to observe retinal morphology.RESULTS(1)General condition and behavior:The RPG group showed a continuous decline in scores for fur color,limb muscles,movement,and mental status,with lower body weight and body temperature than PG group(tweight=6.014,P=0.000;ttemperature=3.125,P=0.005),and the differences were statistically significant.(2)Spleen and thymus indices:There was no statistically significant difference in thymus and spleen indices among the three groups(P>0.05).(3)Serum D-xylose and α-amylase:Serum D-xylose levels in the PG and RPG groups were lower than those in the CG group(tPG=11.270,tRPG=17.990,both P=0.000),and the RPG group was lower than the PG group(t= 6.720,P=0.000),with statistically significant differences.There was no statistically significant difference in serum α-amylase among the three groups(P>0.05).(4)Fundus CNV:The CNV exudation area of rats in the RPG group at seven and 14 days was higher than that in the PG group(t7 d=2.217,t14 d=2.210,both P=0.032),and the grayscale value was higher than that in the PG group(t7 d=3.448,P=0.001;t14 d=2.215,P=0.032),with statistically significant differences.(5)Retinal structural morphology:Rats in CG group showed a complete and clear retinal structure,while those in PG and RPG groups showed retinal structure damage with bleeding,edema,and neovascularization,with more significant changes in RPG group than in PG group.(6)Retinal lesion area:The average lesion area in RPG group was higher than that in PG group,with statistically significant differences(t=4.645,P=0.000).CONCLUSIONS Intragastric administration of rhubarb decoction combined with retinal laser photocoagulation can successfully establish a rat model of spleen deficiency syndrome combined with wAMD.
关键词
脾虚证/湿性年龄相关性黄斑变性/脉络膜新生血管/病证结合/动物模型Key words
spleen deficiency syndrome/wet age-related macular degeneration/choroidal neovascularization/combination of disease and syndrome/animal model引用本文复制引用
基金项目
国家自然科学基金(81973912)
国家自然科学基金(82274589)
首都卫生发展科研专项(2020-2-4183)
中国中医科学院委托项目(ZZ13-036-1)
中国中医药循证医学中心业务研究室主任专项(2020YJSZX-2)
出版年
2024
国家科技期刊平台
NSTL
维普
万方数据