Angiotensin II as a Morphogenic Cytokine Stimulating Fibrogenesis of Human Tenon's Capsule Fibroblasts
Angiotensin II as a Morphogenic Cytokine Stimulating Fibrogenesis of Human Tenon's Capsule Fibroblasts
复制标题
血管紧张素 II 作为形态发生细胞因子刺激人腱囊成纤维细胞的纤维形成。
DOI：
10.1167/iovs.14-15301
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发表时间：
2015-02-01
影响因子：
4.4
通讯作者：
Xiao, Yiqin
中科院分区：
文献类型：
--
作者：
Shi, Huimin;Zhang, Yuyan;Xiao, Yiqin
目的。检查血管紧张素II（ANG II）及其I型和II型受体（AT1R，AT2R）在小梁切除术后兔囊胶囊成纤维细胞中的表达，并研究Ang II对培养的人类田纳剂胶囊的影响增殖，迁移，表型过渡和细胞外基质（ECM）综合。在兔子中，通过免疫组织化学评估了小梁切除术后ang II，AT1R和AT2R眼睛囊胶囊成纤维细胞的表达。 ELISA评估了幽默和血浆中的ANG II水平。从接受白内障手术的患者获得的培养的HTF接受了ANG II，TGF-beta 1或媒介物控制的治疗。分别通过细胞计数和Transwell分析和伤口刮擦测定法评估细胞增殖和迁移。通过蛋白质印迹和免疫荧光测量α-光滑肌肉肌动蛋白（α-SMA）和纤连蛋白（FN）的蛋白质表达。通过实时pcr。测量α-SMA和FN的Messenger RNA表达式。在兔子中，ANG II和AT1R的表达与手术后的1天相比增加，而AT2R的表达从7天开始增加。在培养的HTF中，ANG II显着促进了细胞增殖和迁移（p <0.05）。有趣的是，10（-7）M Ang II的作用比较高浓度更为突出（10（-5）M; P <0.05）。 ANG II还显着诱导了α-SMA和FN的表达，这表明表型过渡到肌纤维细胞。我们的结果表明，小梁切除术会改变齿轮囊成纤维细胞中ANG II及其受体的水平，并且ANG II增加了HTFS增殖，迁移和表型过渡，这表明ANG II可能在小梁切除术后在伤口愈合中起作用。
PURPOSE
To examine the expression of Angiotensin II (Ang II) and its type I, and type II receptors (AT1R, AT2R) in rabbit Tenon's capsule fibroblasts after trabeculectomy, and to investigate the effects of Ang II on cultured human Tenon's capsule fibroblasts (HTFs) proliferation, migration, phenotype transition, and extracellular matrix (ECM) synthesis.
METHODS
In the rabbit, expression of Ang II, AT1R, and AT2R in Tenon's capsule fibroblasts of eyes after trabeculectomy was evaluated by immunohistochemistry. Ang II levels in aqueous humor and plasma were assessed by ELISA. Cultured HTFs, obtained from patients undergoing cataract surgery, were treated with Ang II, TGF-β1, or vehicle control. Cell proliferation and migration were evaluated by Cell Counting Kit-8 and Transwell assay, and wound scratch assay, respectively. Protein expressions of α-smooth muscle actin (α-SMA) and fibronectin (FN) were measured by Western blot and immunofluorescence. Messenger RNA expressions of α-SMA and FN were measured by real-time PCR.
RESULTS
In the rabbit, the expression of Ang II and AT1R increased from 1 day after surgery while AT2R increased from 7 days. In cultured HTFs, Ang II promoted cell proliferation and migration significantly (P < 0.05). Interestingly, the effect of 10(-7) M Ang II was more prominent than higher concentrations (10(-5) M; P < 0.05). Ang II also markedly induced the expression of α-SMA and FN, suggesting a phenotypic transition to myofibroblasts.
CONCLUSIONS
Our results show that trabeculectomy alter the levels of Ang II and its receptors in Tenon's capsule fibroblasts, and that Ang II increase HTFs proliferation, migration, and phenotype transition, suggesting that Ang II may play a role in wound healing after trabeculectomy.