Department of Ophthalmology, Eye & ENT Hospital, Fudan University, Shanghai, China.
Eye Institute, Eye & ENT Hospital, Fudan University, Shanghai, China.
J Cell Mol Med. 2021 Jan;25(1):147-160. doi: 10.1111/jcmm.15888. Epub 2020 Nov 18.
We investigated how Src-homology 2-domain phosphatase-1 (SHP-1) regulates the inflammatory response in endotoxin-induced uveitis (EIU), and the signalling pathways involved. One week after intravitreal injection of short hairpin RNA targeting SHP-1 or SHP-1 overexpression lentivirus in rats, we induced ocular inflammation with an intravitreal injection of lipopolysaccharide (LPS). We then assessed the extent of inflammation and performed full-field electroretinography. The concentrations and retinal expression of various inflammatory mediators were examined with enzyme-linked immunosorbent assays and Western blotting, respectively. SHP-1 overexpression and knockdown were induced in Müller cells to study the role of SHP-1 in the LPS-induced inflammatory response in vitro. Retinal SHP-1 expression was up-regulated by LPS. SHP-1 knockdown exacerbated LPS-induced retinal dysfunction and increased the levels of proinflammatory mediators in the retina, which was abrogated by a c-Jun N-terminal kinase (JNK) inhibitor (SP600125). SHP-1 overexpression had the opposite effects. In Müller cells, the LPS-induced inflammatory response was enhanced by SHP-1 knockdown and suppressed by SHP-1 overexpression. SHP-1 negatively regulated the activation of the transforming growth factor-β-activated kinase-1 (TAK1)/JNK pathway, but not the nuclear factor-κB pathway. These results indicate that SHP-1 represses EIU, at least in part, by inhibiting the TAK1/JNK pathway and suggest that SHP-1 is a potential therapeutic target for uveitis.
我们研究了Src 同源 2 结构域磷酸酶 1(SHP-1)如何调节内毒素诱导的葡萄膜炎(EIU)中的炎症反应,以及涉及的信号通路。在大鼠玻璃体内注射靶向 SHP-1 的短发夹 RNA 或 SHP-1 过表达慢病毒一周后,我们用玻璃体内注射脂多糖(LPS)诱导眼内炎症。然后,我们评估了炎症的程度并进行了全视野视网膜电图检查。用酶联免疫吸附试验和 Western blot 分别检测各种炎症介质的浓度和视网膜表达。在体外研究 SHP-1 在 LPS 诱导的炎症反应中的作用时,诱导 Müller 细胞中 SHP-1 的过表达和敲低。LPS 上调视网膜 SHP-1 表达。SHP-1 敲低加重了 LPS 诱导的视网膜功能障碍,并增加了视网膜中促炎介质的水平,而 c-Jun N 端激酶(JNK)抑制剂(SP600125)则消除了这种作用。SHP-1 过表达则产生相反的效果。在 Müller 细胞中,SHP-1 敲低增强了 LPS 诱导的炎症反应,而 SHP-1 过表达则抑制了该反应。SHP-1 负调控转化生长因子-β激活激酶 1(TAK1)/JNK 通路的激活,但不调控核因子-κB 通路。这些结果表明,SHP-1 通过抑制 TAK1/JNK 通路来抑制 EIU,至少部分如此,并表明 SHP-1 是葡萄膜炎的潜在治疗靶点。