The mechanism of repressive effects of transthyretitin on the growth of human retinal microvascular endothelial cells under high glucose and hypoxia environment_Chinese Journal of Ocular Fundus Diseases

Authors：

Yin Xiaowen , Shao Jun , Zou Jian , Yin Ying , Hu Yaling , Li Zheng , Zong Da , Chen Xuan , Zhuang Miao , Tan Chengye , Liu Yan ,  Yao Yong

Department of Ophthalmology, Wuxi People's Hospital Affiliated to Nanjing Medical University, Wuxi 214023, China;

Corresponding author：

Yao Yong, Email: pard1@126.com

Keywords：

Retinal Vessels/cytology; Endothelial cells/physiology; Prealbumin/drug effects

DOI：

10.3760/cma.j.issn.1005-1015.2017.05.019

Video：

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Objective To explore repressive effects of transthyretitin (TTR) on the growth of human retinal endothelial cells (hREC) under high glucose and hypoxia environment.Methods hRECs were divided into 8 groups, including normal glucose group (5.5 mmol/L glucose), hypoxia group, high glucose group (25.0 mmol/L glucose), high glucose and hypoxia group, normal glucose group+TTR, normal glucose and hypoxia group+TTR, high glucose group+TTR, high glucose and hypoxia group+TTR. Flow cytometry was used to analyze cellular apoptosis. The expression level of Akt, p-Akt, eNOS, Bcl-2 and Bax protein were measured by Western blot.Results Hypoxia could induce apoptosis as the apoptosis rate of normal and hypoxia group was higher than normal group (χ²=25.360, P＜0.05), high glucose and hypoxia group was higher that high glucose group (χ²=17.400, P＜0.05). The cell apoptosis rate of high glucose and hypoxia group+TTR were increased significantly as compared with high glucose and hypoxia group (χ²=9.900, P＜0.05). There was no statistically significant difference on the cell apoptosis rate between normal group and high glucose group, normal group+TTR and normal group, high glucose group+TTR and high glucose group, normal and hypoxia group+TTR and normal and hypoxia group (P＞0.05). Western blot showed that the expression of Akt did not change significantly in all eight groups（F=2.450, P＞0.05). Compared to normal group, the expression of p-Akt, eNOS, Bcl-2 in normal and hypoxia group were decreased (t=9.406, 5.306, 4.819), and the expression of Bax (t=−4.503) was increased (P＜0.05). Compared to high glucose group, same trend was found in high glucose and hypoxia group (t=8.877, 7.723, 6.500, −14.646; P＜0.05). The expression of p-Akt in normal and hypoxia group+TTR was higher than normal and hypoxia group (t=−5.024, P＜0.05) , but there was no difference on the expression of eNOS, Bcl-2, Bax between these two groups (t=−2.235, −2.656, −0.272; P＞0.05). Compared to high glucose and hypoxia group, the expression of p-Akt and Bcl-2 in high glucose and hypoxia group+TTR were decreased (t=4.355, 4.308; P＜0.05), the expression of Bax was increased (t=−4.311, P＜0.05), and there was no difference on the expression of eNOS between these two groups (t=−1.590, P＞0.05). There was no statistically significant difference in the expression of p-Akt, eNOS, Bcl-2, Bax between high glucose group and normal group (t=−3.407, −4.228, −4.302, −2.076; P＞0.05), normal group+TTR and normal group (t=−4.245, −4.298, −2.816, −1.326; P＞0.05), high glucose group+TTR and high glucose group (t=4.016, −0.784, 0.707, −0.328; P＞0.05).Conclusion Under high glucose and hypoxia, transthyretitin suppress the growth of hREC through Akt/Bcl-2/Bax, but not Akt/eNOS signaling pathway.

Citation： Yin Xiaowen, Shao Jun, Zou Jian, Yin Ying, Hu Yaling, Li Zheng, Zong Da, Chen Xuan, Zhuang Miao, Tan Chengye, Liu Yan, Yao Yong. The mechanism of repressive effects of transthyretitin on the growth of human retinal microvascular endothelial cells under high glucose and hypoxia environment. Chinese Journal of Ocular Fundus Diseases, 2017, 33(5): 523-526. doi: 10.3760/cma.j.issn.1005-1015.2017.05.019 Copy

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1. Buxbaum JN, Reixach NL. Transthyretin: the servant of many masters[J]. Cell Mol Life Sci, 2009, 66(19): 3095-3101. DOI: 10.1007/s00018-009-0109-0.
2. Hamilton JA, Benson MD. Transthyretin: a review from a structural perspective[J]. Cell Mol Life Sci, 2001, 58(10): 1491-1521. DOI: 10.1007/PL00000791.
3. Oshima Y, Deering T, Oshima S, et al. Angiopoietin-2 enhances retinal vessel sensitivity to vascular endothelial growth factor[J]. J Cell Physiol, 2004, 199(3): 412-417. DOI: 10.1002/jcp.10442.
4. 庄淼, 邵珺, 谭澄烨, 等. 转甲状腺素蛋白对视网膜微血管内皮细胞生物学行为的影响[J]. 中华眼底病杂志, 2015, 31(4): 368-370. DOI: 10.3760/cma.j.issn.1005-1015.2015.04.014.Zhuang M, Shao J, Tan CY, et al. Effects of transthyretin on biological behavior of retinal microvascular epithelial cell[J]. Chin J Ocul Fundus Dis, 2015, 31(4): 368-370. DOI: 10.3760/cma.j.issn.1005-1015.2015.04.014.
5. 邵珺, 姚勇. 高糖缺氧环境下转甲状腺素蛋白对视网膜血管内皮细胞的影响[J]. 中华眼底病杂志, 2016, 32(2): 159-162. DOI: 10.3760/cma.j.issn.1005-1015.2016.02.011.Shao J, Yao Y. Transthyretinin repress retinal microvascular endothelial cells under high glucose and hypoxia environment[J]. Chin J Ocul Fundus Dis, 2016, 32(2): 159-162. DOI: 10.3760/cma.j.issn.1005-1015.2016.02.011.
6. Shao J, Yao Y. Transthyretin represses neovascularization in diabetic retinopathy[J]. Mol Vis, 2016, 22: 1188-1197.
7. Li X, Zarbin M, Bhagat N, et al. Anti-vascular endothelial growth factor injections: the new standard of care in proliferative diabetic retinopathy[J]. Dev Ophthalmol, 2017, 60: 131-142. DOI: 10.1159/000459699.
8. Chen X, Chen Q, Wang L, et al. Ghrelin induces cell migration through GHSR1a-mediated PI3K/Akt/eNOS/NO signaling pathway in endothelial progenitor cells[J]. Metabolism, 2013, 62(5): 743-752. DOI: 10.1016/j.metabol.2012.09.014.
9. Li J, Xuan W, Yan R, et al. Remote preconditioning provides potent eardioproteetion via PBK/Akt activation and is associated with nuclear accumulation of 3-catenin[J]. Clin Sci(Lond), 2011, 120(10): 451-462. DOI: 10.1042/CS20100466.
10. Kim HS, Kim SY, Kwak YL, et al. Hyperglycemia attenuates myocardial preconditioning of remifentanil[J]. J Surg Res, 2012, 174(2): 231-237. DOI: 10.1016/j.jss.2011.01.018.
11. Uehiyama T, Engelman RM, Maulik N, et al. Role of Akt signaling inmitoehondrial survival pathway triggered by hypoxic preconditioning[J]. Circulation, 2004, 109(24): 3042-3049. DOI: 10.1161/01.CIR.0000130647.29030.90.
12. Kaneda K, Miyamae M, Sugioka S, et al. Sevoflurane enhances ethanol-induced cardiac preconditioning through modulation of protein kinase C, mitochondrial K-ATP channels, and nitric oxide synthase, in guinea pig hearts[J]. Anesth Analg, 2008, 106(1): 9-16. DOI: 10.1213/01.ane.0000297298.93627.36.
13. Wallgren M, Lidman M, Pedersen A, et al. Reconstitution of the anti-apoptosis Bcl-2 protein into lipid membranes and biophysical evidence for its detergent-driven association with the pro-apoptotic Bax protein[J/OL]. PLoS One, 2013, 8(4): 61452[2013-04-23]. 10.1371/journal.pone.0061452">http://dx.plos.org/10.1371/journal.pone.0061452. DOI: 10.1371/journal.pone.0061452.
14. Siddiqui WA, Ahad A, Ahsan H. The mystery of BCL2 family: Bcl-2 proteins and apoptosis: an update[J]. Arch Toxicol, 2015, 89(3): 289-317. DOI: 10.1007/s00204-014-1448-7.

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The mechanism of repressive effects of transthyretitin on the growth of human retinal microvascular endothelial cells under high glucose and hypoxia environment

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