HMGB1-TLR4和Müller细胞在视网膜血管生成中的作用

doi:10.3969/j.issn.2097-0005.2021.05.007

betway必威登陆网址（betway.com ）学报 ›› 2021, Vol. 42 ›› Issue (5): 362-368.DOI: 10.3969/j.issn.2097-0005.2021.05.007

HMGB1-TLR4和Müller细胞在视网膜血管生成中的作用

孙玉莹¹, 肖欧², 黄春雨³

1.中山大学肿瘤防治中心防癌体检中心,华南肿瘤学国家重点实验室,广东广州 510060;
2.中山大学中山眼科中心,广东广州 510060;
3.中山大学肿瘤防治中心内镜中心,华南肿瘤学国家重点实验室,广东广州 510060

收稿日期:2021-01-11 出版日期:2021-05-25 发布日期:2021-08-31
通讯作者: 黄春雨,E-mail:huangchy@sysucc.org.cn。
作者简介:孙玉莹,主治医师,硕士,主要从事眼底血管相关疾病研究工作,E-mail:sunyy@sysucc.org.cn。
基金资助:
广东省医学科学技术研究基金(A2021369)。

Mechanism effect of HMGB1-TLR4 and Müller cells in retinal angiogenesis

Sun Yuying¹, Xiao Ou², Huang Chunyu³

1. Department of Cancer Prevention, Sun Yat-Sen University Cancer Center, State Key Laboratory of Oncology in South China, Guangzhou 510060, China;
2. Zhongshan Ophthalmological Center, Sun Yat-sen University,Guangzhou 510060,China;
3. Department of Endoscopy, Sun Yat-Sen University Cancer Center, State Key Laboratory of Oncology in South China,Guangzhou 510060,China

Received:2021-01-11 Online:2021-05-25 Published:2021-08-31

摘要/Abstract

摘要： 目的探讨高迁移率族蛋白1-Toll样受体4(HMGB1-TLR4)信号通路及Müller细胞在视网膜新生血管中的作用。方法将小鼠分为3组,分别是常氧组、正常视网膜缺血模型组(OIR组)和TLR4基因敲除OIR(TLR4^-/-OIR)组。通过视网膜FITC-Dextran荧光灌注铺片染色和GS-isolectin B4染色评估新生血管情况。通过免疫荧光染色,观察HMGB1在小鼠视网膜中的表达情况,观察HMGB1和TLR4、TLR4和GFAP的共表达情况。通过PCR检测两组OIR小鼠出生后12天(P12)和出生后17天(P17)时视网膜组织HMGB1 mRNA表达水平。结果通过两种视网膜血管平铺片染色方法观察到P17正常OIR小鼠新生血管面积最大,TLR4^-/-OIR小鼠的新生血管面积相对较少,这提示TLR4基因缺失可以抑制血管的生成。通过免疫荧光染色结果发现在TLR4^-/-OIR小鼠中,HMGB1、TLR4和GFAP的表达均较OIR组减少。PCR检查结果发现P12和P17时,TLR4^-/-OIR小鼠中的HMGB1的表达水平均较正常OIR组低。结论 HMGB1-TLR4可以通过活化Müller细胞促进视网膜新生血管的生成,因此靶向抑制HMGB1-TLR4信号通路,可以降低Müller细胞的活性,抑制视网膜新生血管的生成。

关键词: 高迁移率族蛋白1, Toll样受体4, Müller细胞, 视网膜新生血管, OIR动物模型

Abstract: Objective: To explore the role of HMGB1-TLR4 signal pathway and Müller cells in retinal neovascularization.Methods: The mice were divided into three groups: normoxic group, normal OIR model group and TLR4 gene knockout (TLR4^-/-) OIR model group. Retinal FITC-Dextran fluorescence perfusion and GS-isolectin B4 staining were used to evaluate the neovascularization. Immunofluorescence staining was used to observe the expression of HMGB1 in mouse retina, the co-expressions of HMGB1 and TLR4, and the co-expressions of TLR4 and GFAP. The mRNA expressions of HMGB1 in the retina of OIR mice in P12 and P17 groups were detected by PCR. Results: It was observed that the neovascularization area of P17 normal OIR mice was the largest, while that of TLR4^-/-OIR mice was relatively small, which suggested that TLR4 gene deletion could inhibit angiogenesis. The results of immunofluorescence staining showed that the expressions of HMGB1, TLR4 and GFAP in TLR4^-/-OIR mice were lower than those in normal OIR group. The results of PCR examination showed that the expression levels of HMGB1 in TLR4^-/-OIR mice at P12 and P17 were lower than those in normal OIR group. These results suggested that inhibition of HMGB1-TLR4 signal pathway could weaken the activation of Müller cells and reduce neovascularization.Conclusion: This study explores the mechanism of HMGB1-TLR4 activating Müller cells participating in neovascularization, and reveals the effect of HMGB1-TLR4 on retinal neovascularization by activating Müller cells. It is suggested that blocking HMGB1-TLR4 pathway can weaken the activity of Müller cells and provide a new method for the treatment of retinal neovascularization.

Key words: HMGB1, Toll like receptor 4, Müller cells, retinal neovascularization, oxygen induced retinopathy model

中图分类号:

R774

孙玉莹, 肖欧, 黄春雨. HMGB1-TLR4和Müller细胞在视网膜血管生成中的作用[J]. betway必威登陆网址（betway.com ）学报, 2021, 42(5): 362-368.

Sun Yuying, Xiao Ou, Huang Chunyu. Mechanism effect of HMGB1-TLR4 and Müller cells in retinal angiogenesis[J]. Journal of Shandong First Medical Unversity & Shandong Academy of Medical Sciences, 2021, 42(5): 362-368.

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HMGB1-TLR4和Müller细胞在视网膜血管生成中的作用

Mechanism effect of HMGB1-TLR4 and Müller cells in retinal angiogenesis

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