中华实验眼科杂志
中華實驗眼科雜誌
중화실험안과잡지
CHINESE JOURNAL OF EXPERIMENTAL OPHTHALMOLOGY
2013年
12期
1105-1109
,共5页
转化生长因子-β%骨形态发生蛋白%近视/形觉剥夺性%巩膜
轉化生長因子-β%骨形態髮生蛋白%近視/形覺剝奪性%鞏膜
전화생장인자-β%골형태발생단백%근시/형각박탈성%공막
Transforming growth factor-β/bone morphogenetic protein%Form-deprivation myopia%Sclera
背景 眼球伸长过程伴随着巩膜的广泛重塑,而这种重塑受多种生长因子的调控.以往的研究证实转化生长因子-β(TGF-β)在近视形成和发展过程中发挥作用.骨形态发生蛋白(BMPs)属于TGF-p超家族,其在近视的发生中是否发挥作用及如何发挥作用尚不清楚. 目的 观察豚鼠形觉剥夺性近视(FDM)眼后巩膜中BMPs的表达变化,探讨其在近视后巩膜重塑中的作用.方法 1～2周龄花色豚鼠30只,以随机数字表法随机分为实验组和正常对照组,每组15只.任意选择实验组豚鼠的一侧眼作为实验眼,应用半透明眼罩连续遮盖14 d以建立FDM动物模型,另一侧眼作为对照眼.形觉剥夺前后所有动物眼经检影验光获得屈光度,用A型超声法测量眼轴长度.造模后第15天取豚鼠后巩膜组织,分别用逆转录PCR(RT-PCR)和Western blot法检测各组豚鼠后巩膜中BMPs mRNA及其蛋白的表达. 结果 实验14 d后,豚鼠遮盖眼的屈光度为(-0.48±0.51)D,与对侧眼的(3.22±0.34)D比较,差异有统计学意义(t=-12.814,P=0.000),与正常对照眼的(2.97±0.70)D比较,差异有统计学意义(t=-11.878,P=0.000).豚鼠遮盖眼的眼轴长度为(8.30±0.05)mm,明显长于对侧眼的(8.11±0.06)mm和正常对照组(8.06±0.06) mm,差异均有统计学意义(t=7.230、9.084,均P=0.000).正常豚鼠后巩膜可表达BMP-2、BMP-4、BMP-5 mRNA,豚鼠遮盖眼后巩膜中BMP-2 mRNA和BMP-5 mRNA的相对表达值分别为0.41±0.11和0.65±0.06,较对侧眼的0.62±0.07和0.84±0.03分别下降了34.48％和23.67％,差异均有统计学意义(t=2.838,P=0.017;t2.524,P=0.028);豚鼠遮盖眼后巩膜中BMP-2和BMP-5蛋白表达的相对值分别为0.44±0.06和0.70±0.05,较对侧眼的0.61±0.05和0.82±0.03分别下降了23.42％和15.21％,差异均有统计学意义(t=2.465,P=0.030;t=2.445,P=0.031),而形觉剥夺眼与对侧眼后巩膜中BMP-4 mRNA及其蛋白相对表达量的差异均无统计学意义(mRNA:t=0.704,P=0.460;蛋白:t=0.987,P=0.365).结论 FDM眼后巩膜中BMP-2和BMP-5的表达下调,提示BMPs在实验性近视后巩膜重塑中可能发挥作用.
揹景 眼毬伸長過程伴隨著鞏膜的廣汎重塑,而這種重塑受多種生長因子的調控.以往的研究證實轉化生長因子-β(TGF-β)在近視形成和髮展過程中髮揮作用.骨形態髮生蛋白(BMPs)屬于TGF-p超傢族,其在近視的髮生中是否髮揮作用及如何髮揮作用尚不清楚. 目的 觀察豚鼠形覺剝奪性近視(FDM)眼後鞏膜中BMPs的錶達變化,探討其在近視後鞏膜重塑中的作用.方法 1～2週齡花色豚鼠30隻,以隨機數字錶法隨機分為實驗組和正常對照組,每組15隻.任意選擇實驗組豚鼠的一側眼作為實驗眼,應用半透明眼罩連續遮蓋14 d以建立FDM動物模型,另一側眼作為對照眼.形覺剝奪前後所有動物眼經檢影驗光穫得屈光度,用A型超聲法測量眼軸長度.造模後第15天取豚鼠後鞏膜組織,分彆用逆轉錄PCR(RT-PCR)和Western blot法檢測各組豚鼠後鞏膜中BMPs mRNA及其蛋白的錶達. 結果 實驗14 d後,豚鼠遮蓋眼的屈光度為(-0.48±0.51)D,與對側眼的(3.22±0.34)D比較,差異有統計學意義(t=-12.814,P=0.000),與正常對照眼的(2.97±0.70)D比較,差異有統計學意義(t=-11.878,P=0.000).豚鼠遮蓋眼的眼軸長度為(8.30±0.05)mm,明顯長于對側眼的(8.11±0.06)mm和正常對照組(8.06±0.06) mm,差異均有統計學意義(t=7.230、9.084,均P=0.000).正常豚鼠後鞏膜可錶達BMP-2、BMP-4、BMP-5 mRNA,豚鼠遮蓋眼後鞏膜中BMP-2 mRNA和BMP-5 mRNA的相對錶達值分彆為0.41±0.11和0.65±0.06,較對側眼的0.62±0.07和0.84±0.03分彆下降瞭34.48％和23.67％,差異均有統計學意義(t=2.838,P=0.017;t2.524,P=0.028);豚鼠遮蓋眼後鞏膜中BMP-2和BMP-5蛋白錶達的相對值分彆為0.44±0.06和0.70±0.05,較對側眼的0.61±0.05和0.82±0.03分彆下降瞭23.42％和15.21％,差異均有統計學意義(t=2.465,P=0.030;t=2.445,P=0.031),而形覺剝奪眼與對側眼後鞏膜中BMP-4 mRNA及其蛋白相對錶達量的差異均無統計學意義(mRNA:t=0.704,P=0.460;蛋白:t=0.987,P=0.365).結論 FDM眼後鞏膜中BMP-2和BMP-5的錶達下調,提示BMPs在實驗性近視後鞏膜重塑中可能髮揮作用.
배경 안구신장과정반수착공막적엄범중소,이저충중소수다충생장인자적조공.이왕적연구증실전화생장인자-β(TGF-β)재근시형성화발전과정중발휘작용.골형태발생단백(BMPs)속우TGF-p초가족,기재근시적발생중시부발휘작용급여하발휘작용상불청초. 목적 관찰돈서형각박탈성근시(FDM)안후공막중BMPs적표체변화,탐토기재근시후공막중소중적작용.방법 1～2주령화색돈서30지,이수궤수자표법수궤분위실험조화정상대조조,매조15지.임의선택실험조돈서적일측안작위실험안,응용반투명안조련속차개14 d이건립FDM동물모형,령일측안작위대조안.형각박탈전후소유동물안경검영험광획득굴광도,용A형초성법측량안축장도.조모후제15천취돈서후공막조직,분별용역전록PCR(RT-PCR)화Western blot법검측각조돈서후공막중BMPs mRNA급기단백적표체. 결과 실험14 d후,돈서차개안적굴광도위(-0.48±0.51)D,여대측안적(3.22±0.34)D비교,차이유통계학의의(t=-12.814,P=0.000),여정상대조안적(2.97±0.70)D비교,차이유통계학의의(t=-11.878,P=0.000).돈서차개안적안축장도위(8.30±0.05)mm,명현장우대측안적(8.11±0.06)mm화정상대조조(8.06±0.06) mm,차이균유통계학의의(t=7.230、9.084,균P=0.000).정상돈서후공막가표체BMP-2、BMP-4、BMP-5 mRNA,돈서차개안후공막중BMP-2 mRNA화BMP-5 mRNA적상대표체치분별위0.41±0.11화0.65±0.06,교대측안적0.62±0.07화0.84±0.03분별하강료34.48％화23.67％,차이균유통계학의의(t=2.838,P=0.017;t2.524,P=0.028);돈서차개안후공막중BMP-2화BMP-5단백표체적상대치분별위0.44±0.06화0.70±0.05,교대측안적0.61±0.05화0.82±0.03분별하강료23.42％화15.21％,차이균유통계학의의(t=2.465,P=0.030;t=2.445,P=0.031),이형각박탈안여대측안후공막중BMP-4 mRNA급기단백상대표체량적차이균무통계학의의(mRNA:t=0.704,P=0.460;단백:t=0.987,P=0.365).결론 FDM안후공막중BMP-2화BMP-5적표체하조,제시BMPs재실험성근시후공막중소중가능발휘작용.
Background It is well known that sclera remodeling occurs during axial elongation in myopia under the control of growth hormone or its downstream effectors.The role of transforming growth factor-β (TGF-β) in myopia has been determined in previous studies.Bone morphogenetic protein (BMP) is one of members of the TGF-β superfamily,but if it plays an important role in the genesis and development of myopia is not completely clear.Objective This study was to identify the presence of BMPs in normal guinea pigs sclera and investigate the change of BMPs in the sclera in form-deprivation myopia (FDM) of guinea pigs.Methods Thirty young guinea pigs were randomized into normal control group and experimental group using table of random number.FDM models were established by occluding unilateral eyes of guinea pigs with a translucent lens for 14 days in the experimental group,and the fellow eyes served as the controls.Diopter of all eyes was tested by retinoscopy optometry,and ocular axial length was measured by A-sonography before and after modeling.Posterior sclera tissue of the animals was obtained on 14 days,and the relative expression level of BMPs mRNA and protein were assayed by reverse transcription PCR (RT-PCR) and Western blot.The use and care of the animals complied with ARVO Statement.Results On 14 days after occluding of unilateral eyes,the refraction diopter of the experimental group was (-0.48±0.51) D,and that of the fellow eyes was (3.22 ±0.34) D,showing a significant difference between them (t =-12.814,P =0.000).Also,a significant difference in the diopter was seen between the experimental group and normal control group ([-0.48±0.51]D vs.[2.97±0.70]D,t =-11.878,P=0.000).Axial length was (8.30 ± 0.05) mm in the experimental group,(8.11 ±0.06) mm in the fellow eyes and (8.06±0.06) mm in the normal control group,showing a significant increase in the experimental group compared with the fellow eyes and normal control group (t =7.230,P =0.000 ; t =9.084,P=0.000).The expressions of BMP-2 mRNA,BMP-4 mRNA,BMP-5 mRNA in posterior sclera were detected in the normal guinea pigs.Fourteen days after the induction of myopia,the relative levels of BMP-2 mRNA and BMP-5 mRNA in sclera were 0.41 ± 0.11 and 0.65 ± 0.06 in the experimental eyes,which were significantly lower than 0.62 ± 0.07 and 0.84 ± 0.03 in the fellow eyes with the descent range of 34.48％ and 23.67％ respectively (t=2.838,P=0.017; t=2.524,P=0.028).The relative values of BMP-2 protein and BMP-5 protein were 0.44±0.06 and 0.70±0.05 in the experimental eyes,and those of the fellow eyes were 0.61±0.05 and 0.82±0.03,showing significant decline in the experimental eyes with the lowing range of 23.42％ and 15.21％,respectively (t =2.465,P =0.030;t =2.445,P=0.031).No significant differences were found in the expression of BMP-4 mRNA and protein in posterior sclera between the experimental eyes and the normal control eyes (mRNA:t =0.704,P=0.460;protein:t=0.987,P=0.365).Conclusions The expressions of the BMP-2 and BMP-5 in sclera down-regulate significantly in FDM eyes,which suggest that BMP-2 and BMP-5 participate in sclera remodeling during myopia induction.
