小鼠视网膜激光损伤早期组织功能学变化与单核细胞趋化蛋白动态表达关系的研究
소서시망막격광손상조기조직공능학변화여단핵세포추화단백동태표체관계적연구
Correlation between pathological and functional changes of the retina and the expression of monocyte chemoattractant protein after retinal laser injury in mice
  • 万方数据
    中华眼底病杂志 중화안저병잡지
    CHINESE JOURNAL OF OCULAR FUNDUS DISEASES
    2014年 5期 495-499 ,共5页

    于波%张晓敏%王梅艳%苏畅%蒋元丰%刘勋%李筱荣

    우파%장효민%왕매염%소창%장원봉%류훈%리소영

    视网膜疾病/病因学%视网膜电描记术%趋化因子CCL2%动物实验 視網膜疾病/病因學%視網膜電描記術%趨化因子CCL2%動物實驗
    시망막질병/병인학%시망막전묘기술%추화인자CCL2%동물실험
    Retina diseases/etiology%Electroretinography%Chemokine CCL2%Animal experimentation
    目的 观察小鼠视网膜激光损伤后早期组织功能变化及单核细胞趋化蛋白(MCP-1)表达水平,初步探讨两者之间的关系.方法 116只C57BL/6小鼠随机分为正常组、激光损伤组,均为58只.采用激光光凝方式建立视网膜激光损伤模型.分别于激光光凝后1、3、7d行视网膜电图(ERG)检查.激光光凝后1、3、7d分别处死正常组、激光损伤组小鼠并摘除眼球.行苏木精-伊红染色观察视网膜组织形态变化,实时定量聚合酶链反应(PCR)检测各组小鼠视网膜中MCP-1的基因相对表达情况,蛋白质免疫印迹法(Western blot)检测视网膜中MCP-1的蛋白表达情况.结果 光学显微镜观察结果显示,随激光损伤时间延长,视网膜组织结构损伤进行性加重.ERG检测结果显示,激光损伤后1、3、7d,正常组、激光损伤组小鼠暗适应a(t=6.998、9.594、13.778)、b波(t=12.089、13.310、21.989)振幅比较,差异有统计学意义(P=0.000).激光损伤后1d,正常组、激光损伤组明适应a波振幅比较,差异无统计学意义(t=2.659,P=0.200),b波振幅比较,差异有统计学意义(t=8.844,P=0.000);3、7d,明适应a(t=3.076、7.544)、b波(t=10.418、8.485)振幅比较,差异均有统计学意义(P=0.000).激光损伤后1、3 d(t=3.773)及1、7d(t=5.070)暗适应a波振幅比较,差异有统计学意义(P＜0.01),3、7d比较,差异无统计学意义(t=1.297,P=0.660);b波振幅,1、7d比较,差异有统计学意义(t=4.762,P=0.000);1、3 d(t=2.236)及3、7 d(t=2.526)比较,差异无统计学意义(P=0.120、0.060).明适应a波振幅,1、7d比较,差异有统计学意义(t=2.991,P=0.020);1、3 d(t=0.516)及3、7 d(t=2.475)比较,差异均无统计学意义(P=1.000、0.710);b波振幅,1、3 d(t=3.570)及1、7 d(t=4.989)比较,差异均有统计学意义(P＜0.01),3、7d比较,差异无统计学意义(t=1.419,P=0.070).实时定量PCR检测结果显示,激光损伤后1、3、7d,激光损伤组MCP-1基因相对表达量与正常组比较,差异均有统计学意义(t=14.329、16.861、5.743,P＜0.05).Western blot检测结果显示,激光损伤后1、3、7d,激光损伤组MCP-1蛋白相对表达量与正常组比较,差异均有统计学意义(t=75.068、54.145、14.653,P＜0.05).结论 小鼠视网膜激光损伤后7d内,视网膜组织结构和功能损害进行性加重,且程度与MCP-1表达相关.
    목적 관찰소서시망막격광손상후조기조직공능변화급단핵세포추화단백(MCP-1)표체수평,초보탐토량자지간적관계.방법 116지C57BL/6소서수궤분위정상조、격광손상조,균위58지.채용격광광응방식건립시망막격광손상모형.분별우격광광응후1、3、7d행시망막전도(ERG)검사.격광광응후1、3、7d분별처사정상조、격광손상조소서병적제안구.행소목정-이홍염색관찰시망막조직형태변화,실시정량취합매련반응(PCR)검측각조소서시망막중MCP-1적기인상대표체정황,단백질면역인적법(Western blot)검측시망막중MCP-1적단백표체정황.결과 광학현미경관찰결과현시,수격광손상시간연장,시망막조직결구손상진행성가중.ERG검측결과현시,격광손상후1、3、7d,정상조、격광손상조소서암괄응a(t=6.998、9.594、13.778)、b파(t=12.089、13.310、21.989)진폭비교,차이유통계학의의(P=0.000).격광손상후1d,정상조、격광손상조명괄응a파진폭비교,차이무통계학의의(t=2.659,P=0.200),b파진폭비교,차이유통계학의의(t=8.844,P=0.000);3、7d,명괄응a(t=3.076、7.544)、b파(t=10.418、8.485)진폭비교,차이균유통계학의의(P=0.000).격광손상후1、3 d(t=3.773)급1、7d(t=5.070)암괄응a파진폭비교,차이유통계학의의(P＜0.01),3、7d비교,차이무통계학의의(t=1.297,P=0.660);b파진폭,1、7d비교,차이유통계학의의(t=4.762,P=0.000);1、3 d(t=2.236)급3、7 d(t=2.526)비교,차이무통계학의의(P=0.120、0.060).명괄응a파진폭,1、7d비교,차이유통계학의의(t=2.991,P=0.020);1、3 d(t=0.516)급3、7 d(t=2.475)비교,차이균무통계학의의(P=1.000、0.710);b파진폭,1、3 d(t=3.570)급1、7 d(t=4.989)비교,차이균유통계학의의(P＜0.01),3、7d비교,차이무통계학의의(t=1.419,P=0.070).실시정량PCR검측결과현시,격광손상후1、3、7d,격광손상조MCP-1기인상대표체량여정상조비교,차이균유통계학의의(t=14.329、16.861、5.743,P＜0.05).Western blot검측결과현시,격광손상후1、3、7d,격광손상조MCP-1단백상대표체량여정상조비교,차이균유통계학의의(t=75.068、54.145、14.653,P＜0.05).결론 소서시망막격광손상후7d내,시망막조직결구화공능손해진행성가중,차정도여MCP-1표체상관.
    Objective To investigate the relationship between the pathological and functional changes of the retina and the expression of monocyte chemoattractant protein (MCP)-1 after retinal laser injury in mice.Methods A total of 116 C57BL/6 mice were randomly divided into the normal group (58 mice) and the injured group (58 mice).Retinal laser injuries were induced by Argon ion laser.At 1,3,7 days after laser injury,electroretinogram (ERG) responses were recorded to detect the function of the retina.Hematoxylin and eosin (HE) staining was performed to observe pathological changes.Quantitative real-time polymerase chain reaction (PCR) was performed to detect gene expression of MCP-1.Western blot was used to measure the protein expression of MCP-1.Results HE staining showed a progressive damage of the retinal structure.The results of ERG showed that the differences of dark-adaptive a wave (t =6.998,9.594,13.778) and b wave (t=12.089,13.310,21.989) amplitudes of 1,3 and 7 day post-injury between normal group and injured group were statistically significant (P =0.000).At 1 day post-injury,the differences of light adaptive b wave amplitudes between the two groups were statistically significant (t =8.844,P=0.000).While the differences of light-adaptive a wave amplitudes were not (t=2.659,P=0.200).At 3,7 days post-injury,the differences of a (t=3.076,7.544) and b wave amplitudes (t=10.418,8.485) between the two groups were statistically significant (P=0.000).In dark-adaptive ERG,the differences of a wave amplitudes between 1 day and 3 days (t=3.773),1 day and 7 days (t=5.070) and b wave amplitudes between 1 day and 7 days (t=4.762) were statistically significant (P＜0.01),while the differences of a wave amplitudes between the 3 days and 7 days (t=1.297) and b wave amplitudes between 1dayand 3 days (t=2.236),3 day and 7 days (t=2.526) were not significant (P=0.660,0.120,0.060).In light-adaptive ERG,the differences of a wave amplitudes between 1 day and 7 days (t=2.992)and b wave amplitudes between 1 day and 3 days (t=3.570),1day and 7 days (t=4.989) were statistically significant (P＜0.05),while the differences of a wave amplitudes between 1 day and 3 days (t=0.516),the 3 days and 7 days (t=2.475) and b wave amplitudes between 3 days and 7 days (t=1.419) were not significant (P=1.000,0.710,0.070).Quantitative real-time PCR showed that the differences of MCP-1 gene expression at 1,3 and 7 day post-injury between normal group and injured group were statistically significant (t=14.329,16.861,5.743; P＜0.05).Western blot showed that the differences of MCP-1 protein expression at 1,3 and 7 day post-injury between normal group and injured group were statistically significant (t=75.068,54.145,14.653; P＜0.05).Conclusion In the first 7 days after mice retinal laser injury,there are progressive pathological and functional damage of the retina,which might be correlated with MCP-1 expression.
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