摘要
目的 研究血管紧张素 (angiotensin ,Ang )对体外培养的牛眼小梁细胞骨架的动力学影响 ,探讨 Ang 对房水的调节作用及在青光眼发病机制中的意义。方法 (1)牛眼小梁细胞的体外培养 ,应用免疫组化方法 (NSE, 因子相关抗原染色 )细胞鉴定 ,光学及电子透射显微镜对细胞进行形态学及生长特性的观察 ;(2 )以含不同浓度 Ang -的培养液孵育小梁细胞 12～ 2 4h,(终浓度分别是 10 - 5、10 - 6、10 - 7m ol· L- 1 )。免疫组化法 (SABC)对α- sm ooth- actin、tubulin-α染色。结果以计算机图像分析系统分析 ,并统计学检验。结果 牛眼小梁细胞培养成功 ,成活率 80 %～ 10 0 % ,上皮型为主 ,Ang 产生明显的细胞收缩效应 ,actin是收缩的主要效应器。结论 体外培养牛眼小梁细胞技术是研究小梁细胞特性的重要实验技术。 Ang 可以引起小梁细胞的浓度依赖性的收缩 ,提示 Ang可能是房水排出途径的调节因子之一 。
Objective To demonstrate the contractive effect of in vitro bovine trabecular meshwork cell's (TM cells)actin and tubulin which was induced by angiotensin Ⅱ. To probe into the regulative effect of angiotensin Ⅱ to aqueous outflow and the significance of angiotensin Ⅱ in primary open angle glaucomatous(POAG) pathogenesis and mechanism.Methods (1) The bovine trabecular meshwork cultured and cells were identified by immunohistochemistry method(NSE. VIII factor related antigen) . The growing characteritics and morphological feature of cultured primary and passaged cells were observed by inverted microscope and electron microscope;(2) The third passage cells were incubated with different dosages of angiotensin Ⅱ (10 -5 、10 -6 、10 -7 mol·L -1 , final concentration ,diluted by DMEM) for 12h to 24h. Then, α smooth actin and tubulin α were stained using immunohistochemistry method. The results were analyzed by the computer image analytical system and statistical test.Results The cells of bovine trabecular meshwork were cultured successfully .The live rate of trabecular meshwork cells was 80%~100%. Most of them were epithelia type, Angiotensin II administration resulted in dose dependent cell contractions in vitro . The actin playsa critical role in this cell contractile properties.Conclusion Establishing the method of culturing bovine trabecular meshwork cells is an important method for researching the characteristics of TM cells. Angiotensin II can result in TM cells contraction and may be a regulator of POAG.
出处
《眼科新进展》
CAS
2001年第1期24-27,共4页
Recent Advances in Ophthalmology
