血管紧张素Ⅱ对体外培养的牛眼小梁细胞^3H胸腺核苷酸掺入率及胶原合成的影响

目的观测血管紧张素Ⅱ(AngiotensinⅡ,AngⅡ)对体外培养的牛眼小梁细胞(bovine trabecularmeshwork cells,TM cell)3H胸腺核苷酸(3H-thymidine,3H-TdR)掺入率及胶原合成的影响,探讨原发性青光眼的发病机制.方法(1)牛眼小梁细胞的体外培养应用免疫组化方法(neuronal specific enolas,NSE,Ⅷ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)AngⅡ(1×10-7mol@L-1及1×10-8mol@L-1)以及其Ⅰ型受体(angiotensin receptor type I,AT1)拮抗剂孵育TM细胞,采用检测3H-TdR掺入率的方法了解细胞增殖状态,并用化学方法检测培养液中羟脯氨酸含量,间接推导胶原含量.结果牛眼小梁细胞培养成功,以上皮型为主.AngⅡ明显地增加了牛眼小梁细胞的3H-TdR摄入率,同时培养液中的羟脯氨酸含量亦相应增高.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.AngⅡ可以诱导体外培养的牛眼小梁细胞的细胞增殖率升高,同时促使胶原合成增强.AT1受体拮抗剂可以部分阻断此促增殖效应.眼科学报2001;17209-212.

Effects of angiotensin II on the 3H-TdR incorporation and synthesis of collagen in cultured bovine trabecular meshwork cells]

OBJECTIVE: To investigate the effects of angiotensin II (Ang II) on the cell proliferation and collagen synthesis of bovine trabecular meshwork (TM) cells in vitro. It may probe into the mechanism of primary open-angle glaucoma(POAG). METHODS: (1) The bovine trabecular meshwork cells were cultured and identified by immunohistochemistry method(neuronal specific enolas, NSE, factor related antigan). The growing characteristics and morphological feature of cultured primary and passaged cells were observed by invered microscope and eletron-microscope; (2) Ang II(1 x 10(-7) mol.L-1 and 1 x 10(-8) mol.L-1) and angiotensin receptor type I (AT1) antagonist (losartan) were incubated with cultured TM cells. The cellular proliferation was measured by 3H-thymidine(3H-TdR) incorporation assay and collagen synthesis was indirectly professed though detects the hydroxyproline of medium with chemistry methods. RESULTS: The cells of bovine trabecular meshwork were cultured successfully. Most of them were epithelia type. Ang II increased the TM cells uptake of 3H-TdR and inhibited by losartan partially. Meanwhile the hydroxyproline of medium was increased correspondingly. CONCLUSION: Establishing the method of culturing bovine trabecular meshwork cells is an important method for researching the characteristics of TM cells. Ang II can induce the cell proliferation of bovine TM cells and increase the synthesis of collagen in vitro. The AT1 antagonist may inhibit this cell proliferation effect.