中华实验眼科杂志
中華實驗眼科雜誌
중화실험안과잡지
CHINESE JOURNAL OF EXPERIMENTAL OPHTHALMOLOGY
2015年
2期
123-128
,共6页
沙芳%赵雯%徐超立%王玲%张小燕%吴建峰%毕爱玲%郭大东%毕宏生
沙芳%趙雯%徐超立%王玲%張小燕%吳建峰%畢愛玲%郭大東%畢宏生
사방%조문%서초립%왕령%장소연%오건봉%필애령%곽대동%필굉생
近视/透镜诱导性%γ-氨基丁酸%γ-氨基丁酸受体%视网膜
近視/透鏡誘導性%γ-氨基丁痠%γ-氨基丁痠受體%視網膜
근시/투경유도성%γ-안기정산%γ-안기정산수체%시망막
Myopia/lens-induced%Gamma aminobutyric acid%Gamma aminobutyric acid receptor%Retina
背景 研究表明,抑制性氨基酸神经递质γ-氨基丁酸(GABA)在近视的发生和发展过程中发挥关键作用,GABA及GABA受体拮抗剂对近视发生的抑制作用已被证实,但相关作用机制的研究鲜见报道.目的 探讨负透镜诱导性近视(LIM)豚鼠视网膜中GABA及其受体亚型表达的变化,探讨GABA及GABA受体拮抗剂抑制近视发生的机制.方法 采用随机数字表法将40只3周龄健康三色短毛豚鼠随机分为正常对照组和模型组.正常对照组取右眼作为正常对照眼,模型组右眼佩戴-10 D透镜4周以诱导近视模型作为实验眼,左眼不戴镜作为对照眼.分别于造模前及造模后4周用A型超声法和带状光检影法测量各组豚鼠的眼轴长度和屈光度;于造模后4周断颈法处死豚鼠并分离视网膜,采用高效液相色谱分析(HPLC)法检测豚鼠视网膜中GABA含量的变化,采用SYBR Green Ⅰ实时荧光定量PCR检测豚鼠视网膜中GABA受体mRNA的相对表达量. 结果 造模后4周,模型组实验眼、正常对照眼和模型组对照眼屈光度分别为(-3.39±0.70)、(1.03±0.68)和(0.77±0.64)D,模型组实验眼近视屈光度较正常对照眼和模型组对照眼均明显增大,差异均有统计学意义(t=-16.382、-17.113,均P＜0.05);模型组实验眼、正常对照眼和模型组对照眼眼轴长度分别为(8.62±0.08)、(8.31±0.05)和(8.32±0.05) mm,模型组实验眼眼轴较正常对照眼和模型组对照眼明显延长,差异均有统计学意义(t=10.981、15.838,均P＜0.05);豚鼠正常对照眼、模型组实验眼、模型组对照眼视网膜中GABA含量分别为(14.83±1.20)、(18.63±1.58)和(15.74±0.96) μmol/g,模型组实验眼视网膜中GABA含量明显高于正常对照眼和模型组对照眼,差异均有统计学意义(t=-6.045、6.876,均P＜0.05);模型组实验眼视网膜中GABAA受体、受体GABAC受体mRNA的相对表达量明显高于正常对照眼,差异均有统计学意义(t=-3.211、-2.532,均P＜0.05);模型组实验眼视网膜中GABAA受体、受体GABAC受体mRNA的相对表达量明显高于模型组对照眼,差异均有统计学意义(t=2.512、5.645,均P＜0.05),而3个组间GABAV受体mRNA的相对表达量差异无统计学意义(F=2.676,P＞0.05). 结论 LIM豚鼠视网膜中GABA及GABAA受体mRNA、GABAC受体mRNA表达上调,而GABAB受体mRNA表达无显著变化.
揹景 研究錶明,抑製性氨基痠神經遞質γ-氨基丁痠(GABA)在近視的髮生和髮展過程中髮揮關鍵作用,GABA及GABA受體拮抗劑對近視髮生的抑製作用已被證實,但相關作用機製的研究鮮見報道.目的 探討負透鏡誘導性近視(LIM)豚鼠視網膜中GABA及其受體亞型錶達的變化,探討GABA及GABA受體拮抗劑抑製近視髮生的機製.方法 採用隨機數字錶法將40隻3週齡健康三色短毛豚鼠隨機分為正常對照組和模型組.正常對照組取右眼作為正常對照眼,模型組右眼珮戴-10 D透鏡4週以誘導近視模型作為實驗眼,左眼不戴鏡作為對照眼.分彆于造模前及造模後4週用A型超聲法和帶狀光檢影法測量各組豚鼠的眼軸長度和屈光度;于造模後4週斷頸法處死豚鼠併分離視網膜,採用高效液相色譜分析(HPLC)法檢測豚鼠視網膜中GABA含量的變化,採用SYBR Green Ⅰ實時熒光定量PCR檢測豚鼠視網膜中GABA受體mRNA的相對錶達量. 結果 造模後4週,模型組實驗眼、正常對照眼和模型組對照眼屈光度分彆為(-3.39±0.70)、(1.03±0.68)和(0.77±0.64)D,模型組實驗眼近視屈光度較正常對照眼和模型組對照眼均明顯增大,差異均有統計學意義(t=-16.382、-17.113,均P＜0.05);模型組實驗眼、正常對照眼和模型組對照眼眼軸長度分彆為(8.62±0.08)、(8.31±0.05)和(8.32±0.05) mm,模型組實驗眼眼軸較正常對照眼和模型組對照眼明顯延長,差異均有統計學意義(t=10.981、15.838,均P＜0.05);豚鼠正常對照眼、模型組實驗眼、模型組對照眼視網膜中GABA含量分彆為(14.83±1.20)、(18.63±1.58)和(15.74±0.96) μmol/g,模型組實驗眼視網膜中GABA含量明顯高于正常對照眼和模型組對照眼,差異均有統計學意義(t=-6.045、6.876,均P＜0.05);模型組實驗眼視網膜中GABAA受體、受體GABAC受體mRNA的相對錶達量明顯高于正常對照眼,差異均有統計學意義(t=-3.211、-2.532,均P＜0.05);模型組實驗眼視網膜中GABAA受體、受體GABAC受體mRNA的相對錶達量明顯高于模型組對照眼,差異均有統計學意義(t=2.512、5.645,均P＜0.05),而3箇組間GABAV受體mRNA的相對錶達量差異無統計學意義(F=2.676,P＞0.05). 結論 LIM豚鼠視網膜中GABA及GABAA受體mRNA、GABAC受體mRNA錶達上調,而GABAB受體mRNA錶達無顯著變化.
배경 연구표명,억제성안기산신경체질γ-안기정산(GABA)재근시적발생화발전과정중발휘관건작용,GABA급GABA수체길항제대근시발생적억제작용이피증실,단상관작용궤제적연구선견보도.목적 탐토부투경유도성근시(LIM)돈서시망막중GABA급기수체아형표체적변화,탐토GABA급GABA수체길항제억제근시발생적궤제.방법 채용수궤수자표법장40지3주령건강삼색단모돈서수궤분위정상대조조화모형조.정상대조조취우안작위정상대조안,모형조우안패대-10 D투경4주이유도근시모형작위실험안,좌안불대경작위대조안.분별우조모전급조모후4주용A형초성법화대상광검영법측량각조돈서적안축장도화굴광도;우조모후4주단경법처사돈서병분리시망막,채용고효액상색보분석(HPLC)법검측돈서시망막중GABA함량적변화,채용SYBR Green Ⅰ실시형광정량PCR검측돈서시망막중GABA수체mRNA적상대표체량. 결과 조모후4주,모형조실험안、정상대조안화모형조대조안굴광도분별위(-3.39±0.70)、(1.03±0.68)화(0.77±0.64)D,모형조실험안근시굴광도교정상대조안화모형조대조안균명현증대,차이균유통계학의의(t=-16.382、-17.113,균P＜0.05);모형조실험안、정상대조안화모형조대조안안축장도분별위(8.62±0.08)、(8.31±0.05)화(8.32±0.05) mm,모형조실험안안축교정상대조안화모형조대조안명현연장,차이균유통계학의의(t=10.981、15.838,균P＜0.05);돈서정상대조안、모형조실험안、모형조대조안시망막중GABA함량분별위(14.83±1.20)、(18.63±1.58)화(15.74±0.96) μmol/g,모형조실험안시망막중GABA함량명현고우정상대조안화모형조대조안,차이균유통계학의의(t=-6.045、6.876,균P＜0.05);모형조실험안시망막중GABAA수체、수체GABAC수체mRNA적상대표체량명현고우정상대조안,차이균유통계학의의(t=-3.211、-2.532,균P＜0.05);모형조실험안시망막중GABAA수체、수체GABAC수체mRNA적상대표체량명현고우모형조대조안,차이균유통계학의의(t=2.512、5.645,균P＜0.05),이3개조간GABAV수체mRNA적상대표체량차이무통계학의의(F=2.676,P＞0.05). 결론 LIM돈서시망막중GABA급GABAA수체mRNA、GABAC수체mRNA표체상조,이GABAB수체mRNA표체무현저변화.
Background Researches showed that gamma aminobutyric acid (GABA),an inhibitory amino acid neurotransmitter,plays a critical role in myopia refractive development.The inhibitory roles of GABA and its receptors antagonists have been confirmed.However,the study on whether GABA participates in the mechanisms of lens-induced myopia (LIM) in guinea pigs is rare.Objective The aim of this study was to explore the alteration of expression of GABA and its three receptor subtypes in the retinas of guinea pigs with LIM.Methods Threeweek-old healthy guinea pigs were randomized into the normal control group and the model group.The right eyes of the guinea pigs in the normal control group served as the normal control eyes,and the right eyes of the guinea pigs in the model group monocularly fitted with-10 D lens as the experimental eyes,and the fellow eyes of the model group were used as the self-control eyes.The refraction and axial length of eyes were measured by A-sonography and streak retinoscopy before and 4 weeks after modeling.The guinea pigs were sacrificed at the fourth week after modeling,and the retinas were isolated.GABA content in the retinas was assayed by high performance liquid chromatography (HPLC) and the relative expression levels of subtypes of GABA receptors were detected by real-time PCR.All the results were compared among the groups.All experimental protocols and animal handling procedures were approved by the Animal Care and Use Committee of Shandong University of Traditional Chinese Medicine,and were in accordance with the statement of the Association for Research in Vision and Ophthalmology for the use of animals.Results At the fourth week after modeling,the mean diopter of the model eyes was (-3.39±0.70)D,which was significantly higher than (1.03±0.68) D of the normal control eyes and (0.77 ±0.64) D of the self-control eyes in the model group (t =-16.382,-17.113,both at P＜0.05).The mean axial length of the experimental eyes was (8.62±0.08)mm,which was increased in comparison with (8.31 ±0.05) mm of the normal control eyes and (8.32± 0.05) mm of the self-control eyes in the model group (t =10.981,15.838,both at P＜0.05).The GABA contents were (14.83 ± 1.20),(18.63±1.58) and (15.74±0.96)μmol/g in the normal control eyes,experimental eyes and self-control eyes in the model group,showing significant differences between the experimental eyes and normal control eyes or self-control eyes (t =-6.045,6.876,both at P＜0.05).The relative expression levels of GABAA and GABAc receptors mRNA in the retinas were significantly increased in the experimental eyes compared with the normal control eyes (t =-3.211,-2.532,both at P＜0.05) and the self-control eyes in the model group (t=2.512,5.645,both at P＜0.05).However,no significant difference was seen in the relative expression of GABAB receptors mRNA among the three groups (F =2.676,P ＞ 0.05).Conclusions The expressions of GABA as well as the mRNA of GABAA and GABAc receptor up-regulate,but there is no obvious alteration in the expression of mRNA of GABAB receptor in the LIM eyes.
