[1]杜文文,张凤妍,杜珊珊,等.Rho/Rock信号转导通路中的关键因子RhoA、Rock I、p-LMC在后发性白内障动物模型中的表达[J].眼科新进展,2016,36(2):105-109.[doi:10.13389/j.cnki.rao.2016.0029]
 DU Wen-Wen,ZHANG Feng-Yan,DU Shan-Shan,et al.Expressions of key factors ( RhoA, Rock I, p-LMC) of Rho/Rock signaling pathway in animal model of posterior capsule opacification[J].Recent Advances in Ophthalmology,2016,36(2):105-109.[doi:10.13389/j.cnki.rao.2016.0029]
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Rho/Rock信号转导通路中的关键因子RhoA、Rock I、p-LMC在后发性白内障动物模型中的表达
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《眼科新进展》[ISSN:1003-5141/CN:41-1105/R]

卷:
36卷
期数:
2016年2期
页码:
105-109
栏目:
实验研究
出版日期:
2016-02-05

文章信息/Info

Title:
Expressions of key factors ( RhoA, Rock I, p-LMC) of Rho/Rock signaling pathway in animal model of posterior capsule opacification
作者:
杜文文张凤妍杜珊珊邵敬芝祁颖
450052 河南省郑州市,郑州大学第一附属医院眼科
Author(s):
DU Wen-Wen ZHANG Feng-Yan DU Shan-ShanSHAO Jing-Zhi QI Ying
Department of Ophthalmology, the First Affiliated Hospital of Zhengzhou University , Zhengzhou 450052 , Henan Province , China
关键词:
后发性白内障转化生长因子-βRho/Rock信号转导通路
Keywords:
posterior capsule opacification transfer growth factor-β Rho/Rock signaling pathway
DOI:
10.13389/j.cnki.rao.2016.0029
文献标志码:
A
摘要:
目的 建立SD大鼠后囊膜混浊(posteriorcapsularopacification,PCO)动物模型,检测Rho/Rock通路中的关键因子RhoA、Rho激酶Ⅰ(Rhoassociatedcoiledcoilformingproteinkinase-Ⅰ,RockⅠ)和肌球蛋白轻链(phosphorylatedmyosinlightchain,p-MLC)在PCO中的表达。探讨Rho/Rock通路在PCO形成过程中的作用。方法 选用健康SD大鼠30只,随机分为对照组(0d)和实验组(3d、7d、14d、28d),每个时间点各6只。所有大鼠(右眼)均行晶状体囊外摘出术,显微镜下观察不同时间点术眼前节的炎症反应及PCO发生情况。ELISA法检测不同时间点房水中TGF-β的表达。处死大鼠后摘除眼球行组织病理学检查,免疫组织化学和WesternBlot法检测RhoA、RockⅠ、p-MLC在晶状体后囊上的表达。结果 随着术后观察时间的推移,PCO混浊程度逐渐增加,HE染色显示晶状体上皮细胞不同程度的增生、移行。对照组术后0d、3d、7d、14d、28d房水中TGF-β的表达量分别为(37.28±1.31)μg·L-1、(40.29±3.17)μg·L-1、(42.12±3.45)μg·L-1、(60.42±4.75)μg·L-1、(68.69±8.06)μg·L-1,实验组各时间点与对照组相比,差异有统计学意义(F=137.158,P<0.05);不同时间点两两相比,差异均有统计学意义(均为P<0.05)。术后0d、3d、7d、14d和28d晶状体后囊膜上RhoA蛋白相对表达量(RhoA/GAPDH)分别为1.00±0.00、1.49±0.01、1.57±0.03、1.77±0.02和1.97±0.02,实验组各时间点与对照组相比,差异有统计学意义(F=94.546,P<0.05);不同时间点两两相比,差异均有统计学意义(均为P<0.05)。RockⅠ蛋白的表达量(RockⅠ/GAPDH)分别为1.00±0.00、0.35±0.12、1.42±0.14、1.78±0.09和1.95±0.14,实验组各时间点与对照组相比,差异有统计学意义(F=30.942,P<0.05);不同时间点两两相比,差异均有统计学意义(均为P<0.05)。p-MLC蛋白的相对表达量(p-MLC/GAPDH)分别为1.00±0.00、1.44±0.08、1.47±0.13、1.63±0.10和2.12±0.20,实验组各时间点与对照组相比,差异有统计学意义(F=34.313,P<0.05);不同时间点两两相比,差异均有统计学意义(均为P<0.05)。结论 RhoA、RockⅠ、p-MLC在PCO形成过程中表达逐渐增多,Rho/Rock信号转导通路可能与PCO的发生、发展有关。
Abstract:
Objective To establish animal model of posterior capsular opacification ( PCO) in SD rats and detect the expressions of key factors of Rho/Rock pathway in PCO tissue. including RhoA. Rho associated coiledcoil forming protein kinase- I ( Rock I ) and phosphorylated myosin light chain ( p-MLC) . Methods Thirty healthy SD rats were randomly divided into control group (0 day) and experimental group ( 3 days ,7 days , 14 days,28 days ) . Extracapsular extraction was performed in all the rats ( right eyes) . Anterior segment inflammation and development of PCO were observed by slit lamp microscope after surgery. The expression of transfer growth factor-¦Â ( TGF-¦Â ) in aqueous humor was detected by ELISA. Then the rats were killed and the right eyes were removed for histopathological exanunation. Immunohistochemistry and Western Blot were performed to detect the expression of RhoA,Rock I ,p-MLC in posterior capsules. Results The posterior capsular opacification developed gradually after surgery. HE staining showed different degrees of proliferation and migration of lens epithelial cells. After operation, the content of TGF-[3 in the aqueous humor at o day, 3 days,7 days, 14 days,28 days were ( 37. 28 + 1. 31) Vg . L-l , ( 40. 29 +3. 17 ) yg . L-l , ( 42. 12 + 3. 45 ) yg . L ’1 , ( 60. 42 +4. 75 ) Vg . L ’I , (68. 69 +8. 06) Vg . L ’l , respectively , the difference of TGF-¦Â content was statistically sigruficant between the experimental group and control group (F = 137. 158 .P = 0. 000 ) . and the TGF-¦Â content was statistically significant among different experiment groups ( all P < 0. 05 ) . The expression of RhoA( RhoA/GAPDH) at o day , 3 days ,7 days , 14 days ,28 days were l. 00 + 0. 00 , 1. 49 + 0. 01 , 1. 57 + 0. 03 . 1. 77 + 0. 02 , 1. 97 + 0. 02 . respectively, the difference of RhoA content was statistically significant between the experimental group and control group ( F = 94. 546 .P = 0. 000 ) . and the RhoA content was statistically significant among different experiment groups ( all P < 0. 05 ) . The expression of Rock I ( Rock I /GAPDH) at o day ,3 days ,7 days , 14 days ,28 days were l. 00 + 0. 00 ,0. 35 + 0. 12 , 1. 42 + 0. 14 , 1. 78 + 0. 09 . 1. 95 + 0. 14 , respectively , the difference of Rock I content was statistically sigruficant between the experimental group and control group ( F = 30. 942 ,P = 0. 000) , and the Rock I content was statistically significant among different experiment groups ( all P < 0. 05 ) . The expression of p-MLC ( p-MLC/GAPDH ) at o day ,3 days ,7 days , 14 days ,28 days were 1. 00 + 0. 00 , 1. 44 + 0. 08 . 1. 47 + 0. 13 . 1. 63 + 0. 10 .2. 12 + 0. 20 , respectively , the difference of p-MLC content was statistically significant between the experimental group and control group ( F = 34. 313 ,P = 0. 000 ) . and the p-MLC content was statistically significant among different experiment groups ( all P < 0. 05 ) . Conclusion The expressions of RhoA.Rock I and p-MLC gradually increase during the development of PCO,indicating that the Rho/Rock signal pathway may be involved in PCO formation.

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备注/Memo

备注/Memo:
河南省科学技术重点攻关项目(编号:142102310053)
更新日期/Last Update: 2016-01-29