The effect of ET1-CTGF mediated pathway on the accumulation of extracellular matrix in the trabecular meshwork and its contribution to the increase in IOP
The effect of ET1-CTGF mediated pathway on the accumulation of extracellular matrix in the trabecular meshwork and its contribution to the increase in IOP
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DOI：
10.1007/s10792-023-02733-y
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发表时间：
2023-05
影响因子：
1.6
通讯作者：
Junming Wang;Y. Rong;Y. Liu;Mengxia Zhu;W. Chen;Zhiqi Chen;Jingmin Guo;C. Deng;A. Manyande;Ping Wang;Hong Zhang;Y. Xiang
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文献类型：
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作者：
Junming Wang;Y. Rong;Y. Liu;Mengxia Zhu;W. Chen;Zhiqi Chen;Jingmin Guo;C. Deng;A. Manyande;Ping Wang;Hong Zhang;Y. Xiang
目的探讨内皮素1（ET 1）在小梁网细胞外基质（ECM）过度积聚中的作用及其在眼压调节中的作用。方法培养人小梁网细胞（HTMCs），分别用ET 1、ET 1 + ETA受体（ETAR）拮抗剂BQ 12 3、ET 1 + ETB受体（ETBR）拮抗剂BQ 788处理。免疫印迹法和免疫荧光法检测纤维连接蛋白（FN）和IV型胶原（Col IV）的表达。通过qRT-PCR研究ET-1对结缔组织生长因子（CTGF）转录水平的时程效应。然后，用反义寡核苷酸序列下调CTGF的转录水平。用ET-1处理HTMCs，Western blotting检测FN和Col IV的表达水平。结果在体外培养的HTMCs中，ET-1处理后FN和Col IV的表达明显增加，ETAR拮抗剂BQ 123可阻断ET-1对HTMCs FN和Col IV表达的影响，而ETBR拮抗剂BQ 788则不能阻断ET-1对HTMCs FN和Col IV表达的影响。ET-1作用48 h后，CTGF mRNA水平显著升高，并达高峰。而CTGF的下调可抑制ET-1对HTMCs FN和Col IV表达的促进作用。在离体模型中，ET-1给药后IOP显著升高，这可以被BQ 123阻断，但不能被BQ 788阻断。结论眼房水中过量的ET-1可通过ETA-CTGF途径导致TM中FN和Col IV的异常积聚，从而导致眼压升高。
PurposeTo investigate the effect of endothelin-1 (ET-1) in excessive accumulation of extracellular matrix (ECM) of the trabecular meshwork (TM) and its role in intraocular pressure (IOP) regulation.MethodsCultured human TM cells (HTMCs) were treated with ET-1, ET-1 + ETA receptor (ETAR) antagonist BQ123, ET-1 + ETB receptor (ETBR) antagonist BQ788. The expressions of fibronectin (FN) and collagen type IV (Col IV) were evaluated by western blotting and immunofluorescence. A time course effect of ET-1 on the transcription level of connective tissue growth factor (CTGF) was investigated by qRT-PCR. Next, the transcription level of CTGF was downregulated by using antisense oligodeoxynucleotide sequence. Then HTMCs were treated with ET-1, and the expression levels of FN and Col IV were evaluated by western blotting. In addition, by using an ex-vivo model of cultured anterior eye segment, we explored the effect of ET-1 on IOP changes and the expressions of FN and Col IV.ResultsIn cultured HTMCs, the expressions of FN and Col IV were significantly increased after ET-1 treatment, which were blocked by the pretreatment of ETAR antagonist BQ123, rather than ETBR antagonist BQ788. Besides, the CTGF mRNA level increased significantly and reached a peak after 48 h of ET-1 treatment. However, the effect of ET-1 on increasing the expressions of FN and Col IV in HTMCs could be inhibited by the downregulation of CTGF. In an ex-vivo model, IOP increased significantly after ET-1 administration, which could be blocked by BQ123 but not by BQ788. Furthermore, elevated expressions of FN and Col IV in TM were observed after ET-1 perfusion, and could be inhibited by BQ123 pretreatment.ConclusionExcessive ET-1 in aqueous humor could lead to the abnormal accumulation of FN and Col IV in TM via the ETA-CTGF pathway, thereby increasing IOP.