背景近年来研究发现,JAK/信号转导及转录激活蛋白3(STAT3)信号转导通路在近视形成和发展中发挥重要作用,AG490是JAK的一种特异抑制剂,因此可抑制JAK2/STAT信号转导,但AG490是否能抑制或延缓近视的进展尚不清楚. 目的探讨玻璃体腔注射酪氨酸激酶选择性抑制剂AG490后STAT3信号通路的活性及其对豚鼠形觉剥夺性近视(FDM)过程中巩膜重塑的影响.方法采用随机数字表法将40只豚鼠随机分为正常对照组、模型对照组、PBS对照组和AG490治疗组,其中模型对照组、PBS对照组和AG490治疗组均用半透明眼罩遮盖右眼4周以制备FDM模型,自遮盖当天向PBS对照组和AG490治疗组豚鼠实验眼玻璃体腔分别注射PBS或AG490各5μl,每2天注射1次,至遮盖结束.分别于实验前及实验后4周检测各组豚鼠双眼的屈光度和眼轴长度,实验4周时摘取实验眼眼球制备巩膜组织切片,采用常规组织病理学检查观察实验眼巩膜的形态学改变,采用免疫组织化学及逆转录PCR(RT-PCR)技术检测各组实验眼巩膜组织中STAT3、p-STAT3、基质金属蛋白酶2(MMP-2)蛋白及其mRNA的表达.结果与正常对照组比较,模型对照组、PBS对照组和AG490治疗组豚鼠实验眼的近视屈光度增加,眼轴明显增长,AG490治疗组眼轴长度分别短于模型对照组和PBS对照组,4个组间差异均有统计学意义(屈光度:F=89.063,P=0.000;眼轴长度:F=96.145,P=0.000).正常对照组豚鼠巩膜组织中STAT3、MMP-2、p-STAT3表达极微弱,AG490治疗组巩膜组织中STAT3、p-STAT3、MMP-2相对表达量(A值)分别为0.064±0.016、0.019±0.002和0.155±0.052,分别低于模型对照组的0.129±0.008、0.071±0.021、0.425±0.004和PBS对照组的0.130±0.004、0.069±0.002、0.421±0.042,差异均有统计学意义(STAT3:t=4.641,9.364,均P＜0.01;p-STAT3:t=4.638、4.488,均P＜0.05;MMP-2:t =9.123、9.029,均P＜0.05),AG490治疗组的表达仍高于正常对照组(t=2.674、2.251、2.682,均P＜0.05).AG490治疗组巩膜组织STAT3 mRNA和MMP-2 mRNA的相对表达量分别为0.295±0.032和0.569±0.019,明显低于模型对照组的0.547±0.015和0.782±0.051以及PBS对照组的0.544±0.015和0.779±0.048,差异均有统计学意义(STAT3 mRNA:t=10.115、11.703,均P＜0.01;MMP-2 mRNA:t=9.218、9.494,均P＜0.01),但与正常对照组STAT3 mRNA水平比较仍有明显上调,差异均有统计学意义(t=2.576、3.565,均P＜0.05). 结论 AG490可一定程度上阻断FDM眼巩膜组织中STAT3信号转导通路的激活,进而调控其下游靶基因MMP-2的转录和表达,减缓FDM眼巩膜的重塑过程,从而一定程度上抑制轴性近视的进展.
揹景近年來研究髮現,JAK/信號轉導及轉錄激活蛋白3(STAT3)信號轉導通路在近視形成和髮展中髮揮重要作用,AG490是JAK的一種特異抑製劑,因此可抑製JAK2/STAT信號轉導,但AG490是否能抑製或延緩近視的進展尚不清楚. 目的探討玻璃體腔註射酪氨痠激酶選擇性抑製劑AG490後STAT3信號通路的活性及其對豚鼠形覺剝奪性近視(FDM)過程中鞏膜重塑的影響.方法採用隨機數字錶法將40隻豚鼠隨機分為正常對照組、模型對照組、PBS對照組和AG490治療組,其中模型對照組、PBS對照組和AG490治療組均用半透明眼罩遮蓋右眼4週以製備FDM模型,自遮蓋噹天嚮PBS對照組和AG490治療組豚鼠實驗眼玻璃體腔分彆註射PBS或AG490各5μl,每2天註射1次,至遮蓋結束.分彆于實驗前及實驗後4週檢測各組豚鼠雙眼的屈光度和眼軸長度,實驗4週時摘取實驗眼眼毬製備鞏膜組織切片,採用常規組織病理學檢查觀察實驗眼鞏膜的形態學改變,採用免疫組織化學及逆轉錄PCR(RT-PCR)技術檢測各組實驗眼鞏膜組織中STAT3、p-STAT3、基質金屬蛋白酶2(MMP-2)蛋白及其mRNA的錶達.結果與正常對照組比較,模型對照組、PBS對照組和AG490治療組豚鼠實驗眼的近視屈光度增加,眼軸明顯增長,AG490治療組眼軸長度分彆短于模型對照組和PBS對照組,4箇組間差異均有統計學意義(屈光度:F=89.063,P=0.000;眼軸長度:F=96.145,P=0.000).正常對照組豚鼠鞏膜組織中STAT3、MMP-2、p-STAT3錶達極微弱,AG490治療組鞏膜組織中STAT3、p-STAT3、MMP-2相對錶達量(A值)分彆為0.064±0.016、0.019±0.002和0.155±0.052,分彆低于模型對照組的0.129±0.008、0.071±0.021、0.425±0.004和PBS對照組的0.130±0.004、0.069±0.002、0.421±0.042,差異均有統計學意義(STAT3:t=4.641,9.364,均P＜0.01;p-STAT3:t=4.638、4.488,均P＜0.05;MMP-2:t =9.123、9.029,均P＜0.05),AG490治療組的錶達仍高于正常對照組(t=2.674、2.251、2.682,均P＜0.05).AG490治療組鞏膜組織STAT3 mRNA和MMP-2 mRNA的相對錶達量分彆為0.295±0.032和0.569±0.019,明顯低于模型對照組的0.547±0.015和0.782±0.051以及PBS對照組的0.544±0.015和0.779±0.048,差異均有統計學意義(STAT3 mRNA:t=10.115、11.703,均P＜0.01;MMP-2 mRNA:t=9.218、9.494,均P＜0.01),但與正常對照組STAT3 mRNA水平比較仍有明顯上調,差異均有統計學意義(t=2.576、3.565,均P＜0.05). 結論 AG490可一定程度上阻斷FDM眼鞏膜組織中STAT3信號轉導通路的激活,進而調控其下遊靶基因MMP-2的轉錄和錶達,減緩FDM眼鞏膜的重塑過程,從而一定程度上抑製軸性近視的進展.
배경근년래연구발현,JAK/신호전도급전록격활단백3(STAT3)신호전도통로재근시형성화발전중발휘중요작용,AG490시JAK적일충특이억제제,인차가억제JAK2/STAT신호전도,단AG490시부능억제혹연완근시적진전상불청초. 목적탐토파리체강주사락안산격매선택성억제제AG490후STAT3신호통로적활성급기대돈서형각박탈성근시(FDM)과정중공막중소적영향.방법채용수궤수자표법장40지돈서수궤분위정상대조조、모형대조조、PBS대조조화AG490치료조,기중모형대조조、PBS대조조화AG490치료조균용반투명안조차개우안4주이제비FDM모형,자차개당천향PBS대조조화AG490치료조돈서실험안파리체강분별주사PBS혹AG490각5μl,매2천주사1차,지차개결속.분별우실험전급실험후4주검측각조돈서쌍안적굴광도화안축장도,실험4주시적취실험안안구제비공막조직절편,채용상규조직병이학검사관찰실험안공막적형태학개변,채용면역조직화학급역전록PCR(RT-PCR)기술검측각조실험안공막조직중STAT3、p-STAT3、기질금속단백매2(MMP-2)단백급기mRNA적표체.결과여정상대조조비교,모형대조조、PBS대조조화AG490치료조돈서실험안적근시굴광도증가,안축명현증장,AG490치료조안축장도분별단우모형대조조화PBS대조조,4개조간차이균유통계학의의(굴광도:F=89.063,P=0.000;안축장도:F=96.145,P=0.000).정상대조조돈서공막조직중STAT3、MMP-2、p-STAT3표체겁미약,AG490치료조공막조직중STAT3、p-STAT3、MMP-2상대표체량(A치)분별위0.064±0.016、0.019±0.002화0.155±0.052,분별저우모형대조조적0.129±0.008、0.071±0.021、0.425±0.004화PBS대조조적0.130±0.004、0.069±0.002、0.421±0.042,차이균유통계학의의(STAT3:t=4.641,9.364,균P＜0.01;p-STAT3:t=4.638、4.488,균P＜0.05;MMP-2:t =9.123、9.029,균P＜0.05),AG490치료조적표체잉고우정상대조조(t=2.674、2.251、2.682,균P＜0.05).AG490치료조공막조직STAT3 mRNA화MMP-2 mRNA적상대표체량분별위0.295±0.032화0.569±0.019,명현저우모형대조조적0.547±0.015화0.782±0.051이급PBS대조조적0.544±0.015화0.779±0.048,차이균유통계학의의(STAT3 mRNA:t=10.115、11.703,균P＜0.01;MMP-2 mRNA:t=9.218、9.494,균P＜0.01),단여정상대조조STAT3 mRNA수평비교잉유명현상조,차이균유통계학의의(t=2.576、3.565,균P＜0.05). 결론 AG490가일정정도상조단FDM안공막조직중STAT3신호전도통로적격활,진이조공기하유파기인MMP-2적전록화표체,감완FDM안공막적중소과정,종이일정정도상억제축성근시적진전.
Background JAK/ signal transducer and activator of transcription 3 (STAT3) signal pathway plays a critical role during the sclera remodeling of experimental myopia.As a tyrosine kinase inhibitor,AG490 can inhibit the activation of this pathway.But whether AG490 plays a role in delaying the development of myopia is not completely clear.Objective This study was to investigate the inhibition of AG490 to activation of STAT3 signaling pathway and the sequential arresting effect on the sclera remodeling in form-deprived myopia (FDM) models.Methods Forty guinea pigs were randomly divided into the normal control group,model control group,PBS control group and AG490 treatment group.FDM models were established by the occlusion of the right eyes of guinea pigs for consecutive 4 weeks using translucent goggles in the model control group,PBS control group and AG490 treated group,and 25 μl PBS or AG490 were respectively injected into vitreous since the first day of modeling in two-day interval till the fourth week in the PBS control group and AG490 treated group.Refractive state and axial length were examined with retinoscopy and A-scan ultrasonography before and 4 weeks after experiment.The experimental eyes were extracted in the fourth week,and the expressions of scleral STAT3,p-STAT3,metal matrix proteinase-2 (MMP-2) proteins and STAT3 mRNA,MMP-2 mRNA were detected by immunocytochemstry and semi-quantitative reverse transcription PCR (RT-PCR) respectively.The use and care of experimental animals followed ARVO.Results Compared to the normal control group,the negative refraction power and axial length were significantly increased in the model control group,PBS control group and AG490 treated group,and the axial length in the AG490 treated group was smaller than those in the model control group and PBS control group,showing significant differences among the 4 groups (refraction:F =89.063,P =0.000;axial length:F =96.145,P =0.000).The expressions of STAT3,MMP-2 and p-STAT3 in scleral tissue were weaker in the normal control group.The expressional values (A values) of STAT3,p-STAT3 and M MP-2 were 0.064 ± 0.016,0.019 ± 0.002 and 0.155 ± 0.052 in the AG490 treated group,which were lower than 0.129±0.008,0.071 ±0.021,0.425 ±0.004 of the model control group and 0.130±0.004,0.069±0.002,0.421 ±0.042 of the PBS control group (STAT3:t =4.641,9.364,both at P＜0.01;p-STAT3:t =4.638,4.488,both at P＜ 0.05;MMP-2:t =9.123,9.029,both at P ＜ 0.05),however,these expressions were still higher than those of the normal control group (t =2.674,2.251,2.682,all at P ＜0.05).The expressional levels (A values) of STAT3 mRNA and MMP-2 mRNA in the AG490 treated group were 0.295±0.032 and 0.569±0.019,which were significantly lower than 0.547±0.015 and 0.782±0.051 in the model group as well as 0.544±0.015 and 0.779±0.048 in the PBS control group (STAT3 mRNA:t =10.115,11.703,both at P＜0.01;MMP-2 mRNA:t =9.218,9.494,both at P＜0.01).The expressional levels (A values) of STAT3 mRNA and MMP-2 mRNA in the AG490 treated group were still higher than those in the normal control group (t=2.576,3.565,both at P＜0.05).Conclusions AG490 can ultimately inhibit the development of axial myopia by arresting the activation of STAT3 signaling pathway in the FDM eyes and further regulating the expression of MMP-2 in sclera and delaying the remodeling of sclera.
