与增殖性糖尿病视网膜病变相关的血管生成素样蛋白8的临床与实验研究
Clinical and experimental study on angiopoietin-like protein 8 associated with proliferative diabetic retinopathy.
作者信息
Dong Chang-Xia, Song Cai-Ping, Zhang Chun-Ping, Dong Mei, Gong Xiu-Rong, Gao He-Ying, Wang Hong
机构信息
Department of Ophthalmology, Yantai Yuhuangding Hospital, Yantai 264000, Shandong Province, China.
Department of Ophthalmology, Weihai Municipal Hospital, Weihai 264200, Shandong Province, China.
出版信息
Int J Ophthalmol. 2017 Dec 18;10(12):1819-1823. doi: 10.18240/ijo.2017.12.05. eCollection 2017.
AIM
To confirm the role of angiopoietin-like protein 8 (Angptl 8) in proliferative diabetic retinopathy (PDR).
METHODS
The sera and aqueous humor of 10 PDR patients and 10 non-diabetic retinopathy (NDR) patients (idiopathic macular hole patients) were collected and the expression of Angptl 8 was detected by enzyme linked immune-sorbent assay (ELISA). Experimental diabetes mice model was induced with streptozotocin. The expression of glycosylated hemoglobin and Angptl 8 in sera was detected. Recombinant Angptl 8 was re-infused into wild type (WT) diabetic mice and spatial frequency threshold and contrast sensitivity were measured. retinal pigment epithelium (RPE) were stimulated by recombinant Angptl 8 for 24h. MMT assay were used to detect cell proliferation. At the same time, qRT-PCR and Western blot was used to measure the expression of proliferation-related factors in PRE cells.
RESULTS
The expression of Angptl 8 was markedly increased in the sera and aqueous humor of PDR patients (=99.02, <0.0001 in sera; =10.42, <0.0001 in aqueous). After successfully establishing the diabetic mice model, we found that glycosylated hemoglobin and Angptl 8 expression levels were increased. Re-infusion of recombinant Angptl 8 into WT diabetic mice could further decrease spatial frequency threshold and contrast sensitivity (<0.01). , RPE cells stimulated by recombinant Angptl 8 could increase the relative absorbance of MMT assay (1.486±0.042 1.000±0.104, <0.05) and proliferating cell nuclear antigen (PCNA) expression (0.55±0.01 0.29±0.03, <0.05). The proliferative effect of Angptl 8 is mainly mediated by increasing the expression of proliferation-activating factors cyclin A1 (4.973±0.205 2.720±0.197, <0.05), cyclin F (5.690±0.219 4.297±0.292, <0.05) and E2F2 (2.297±0.102 1.750±0.146, <0.05), and reducing the expression of proliferation-inhibiting factors cdkn1 (2.370±0.074 3.317±0.135, <0.05) and cdkn2 (4.793±0.065 5.387±0.149, <0.05).
CONCLUSION
The expression of Angptl 8 is increased in PDR, and the increased Angptl 8 can promote proliferation and increase proliferation-related factors.
目的
证实血管生成素样蛋白8（Angptl 8）在增殖性糖尿病视网膜病变（PDR）中的作用。
方法
收集10例PDR患者及10例非糖尿病视网膜病变（NDR）患者（特发性黄斑裂孔患者）的血清和房水，采用酶联免疫吸附测定（ELISA）检测Angptl 8的表达。用链脲佐菌素诱导建立实验性糖尿病小鼠模型，检测血清中糖化血红蛋白和Angptl 8的表达。将重组Angptl 8重新注入野生型（WT）糖尿病小鼠体内，测量空间频率阈值和对比敏感度。用重组Angptl 8刺激视网膜色素上皮（RPE）细胞24小时，采用MMT法检测细胞增殖。同时，用qRT-PCR和蛋白质免疫印迹法检测RPE细胞中增殖相关因子的表达。
结果
PDR患者血清和房水中Angptl 8的表达显著增加（血清中P=99.02，P<0.0001；房水中P=10.42，P<0.0001）。成功建立糖尿病小鼠模型后，发现糖化血红蛋白和Angptl 8表达水平升高。将重组Angptl 8重新注入WT糖尿病小鼠体内可进一步降低空间频率阈值和对比敏感度（P<0.01）。此外，重组Angptl 8刺激的RPE细胞可增加MMT法的相对吸光度（1.486±0.042比1.000±0.104，P<0.05）和增殖细胞核抗原（PCNA）的表达（0.55±0.01比0.29±0.03，P<0.05）。Angptl 8的增殖作用主要通过增加增殖激活因子细胞周期蛋白A1（4.973±0.205比2.720±0.197，P<0.05）、细胞周期蛋白F（5.690±0.219比4.297±0.292，P<0.05）和E2F2（2.297±0.102比1.750±0.146，P<0.05）的表达，以及降低增殖抑制因子cdkn1（2.370±0.074比3.317±0.135，P<0.05）和cdkn2（4.793±0.065比5.387±0.149，P<0.05）的表达来介导。
结论
PDR中Angptl 8表达增加，增加的Angptl 8可促进增殖并增加增殖相关因子。
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