Activation of Sonic Hedgehog Signaling Pathway Regulates Human Trabecular Meshwork Cell Function
Activation of Sonic Hedgehog Signaling Pathway Regulates Human Trabecular Meshwork Cell Function
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DOI：
10.1089/jop.2023.0022
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发表时间：
2023-06-12
影响因子：
2.3
通讯作者：
Li，Hong
中科院分区：
文献类型：
--
作者：
Wang，Xiaochen;Tan，Sisi;Li，Hong
目的：研究Sonic hedgehog（Shh）信号对原代培养的人眼小梁细胞（HTM）的影响。重组Shh（rShh）蛋白和环巴胺分别用于激活和抑制Shh信号通路。进行细胞活力测定以评估rShh对原代HTM细胞活性的影响。还进行了细胞粘附和吞噬作用的功能评估。采用流式细胞术检测凋亡细胞的比例。通过检测纤维连接蛋白（FN）和转化生长因子β 2（TGF-β2）蛋白表达，观察rShh对细胞外基质（ECM）代谢的影响。采用实时荧光定量PCR（RT-PCR）和Western blot方法检测Shh信号通路相关因子GLI家族锌指1（GLI 1）和融合抑制因子（SUFU）的mRNA和蛋白表达。rShh可增强原代HTM细胞的粘附和吞噬能力，减少细胞凋亡。rShh处理的原代HTM细胞FN和TGF-β2蛋白表达增加。rShh上调GLI 1的转录活性和蛋白水平，下调SUFU的转录活性和蛋白水平。相应地，用10 μM浓度的Shh通路抑制剂cyclopamine预处理可部分阻断rShh诱导的GLI 1表达上调。结论：Shh信号通路的激活可通过GLI 1调节原代HTM细胞的功能。Shh信号的调节可能是减轻青光眼细胞损伤的潜在靶点。
Purpose:To investigate the effects of Sonic hedgehog (Shh) signaling on primary human trabecular meshwork (HTM) cells.Methods:Primary HTM cells were isolated from healthy donors and cultured. Recombinant Shh (rShh) protein and cyclopamine were used to activate and inhibit the Shh signaling pathway, respectively. A cell viability assay was performed to assess the effects of rShh on the activity of primary HTM cells. Functional assessment of cell adhesion and phagocytosis was also performed. The proportion of apoptotic cells was examined using flow cytometry. Fibronectin (FN) and transforming growth factor beta2 (TGF-β2) protein were detected to assess the influence of rShh on the metabolism of the extracellular matrix (ECM). Real-time polymerase chain reaction (RT-PCR) and western blot analyses were used to examine mRNA and protein expression of Shh signaling pathway-associated factors GLI Family Zinc Finger 1 (GLI1) and Suppressor of Fused (SUFU).Results:rShh significantly enhanced primary HTM cell viability at a concentration of 0.5 μg/mL. rShh increased the adhesion and phagocytic abilities of primary HTM cells, and decreased cell apoptosis. FN and TGF-β2 protein expression increased in primary HTM cells treated with rShh. rShh upregulated the transcriptional activity and protein levels of GLI1, and downregulated those of SUFU. Correspondingly, the rShh-induced GLI1 upexpression was partially blocked by pretreatment with the Shh pathway inhibitor cyclopamine at a concentration of 10 μM.Conclusions:Activation of Shh signaling can regulate the function of primary HTM cells through GLI1. Regulation of Shh signaling may be a potential target for attenuating cell damage in glaucoma.