Department of Human Anatomy, Chengde Medical University, Chengde, China.
Department of Physiology, Chengde Medical University, Chengde, China.
Pak J Pharm Sci. 2023 Sep;36(5):1457-1466.
The effect of sericin in high glucose (HG)-induced podocyte injury and the mechanisms involving Snai1 and miR-30a-5p were investigated. Bioinformatics and dual-luciferase reporter assay evaluated the relationship of Snai1 with miR-31a-5p. Podocyte injury mouse induced by HG were randomly divided into control (5.5mmol/L D-glucose), HG (30mmol/L D-glucose), HG + Sericin (30mmol/L D-glucose+600μg/ml sericin), miR-30a-5p inhibitor NC (sericin+30mmol/L D-glucose+miR-30a-5p inhibitor negative control) and miR-30a-5p inhibitor groups (sericin+30mmol/L D-glucose+miR-30a-5p inhibitor). The migration ability of podocytes was detected by Transwell assay. The expressions of Snai1, podocin, E-cadherin, FSP-1, ZO-1, α-SMA, Desmin, and miR-30a-5p were assessed with RT-qPCR and Western blot. Snai1 was one direct target of miR-30a-5p. HG group had significantly larger number of migrated podocytes and higher levels of Snai1, FSP-1, α-SMA and Desmin, but significantly lower levels of podocin, ZO-1 and E-cadherin than control and HG + Sericin group. These effects of sericin were reversed by miR-30a-5p inhibitor, as evidenced by increased podocyte migration and increased expressions of Snai1, α-SMA, FSP-1 and Desmin, whereas decreased expressions of podocin, ZO-1 and E-cadherin. Sericin may protect podocytes from damage caused by HG via up-regulating epithelial phenotype markers, down-regulating mesenchymal phenotype markers, and reducing migration of podocytes. The mechanism may be through targeting miR-30a-5p and its target Snai1.
研究了丝胶在高葡萄糖（HG）诱导的足细胞损伤中的作用及其涉及 Snai1 和 miR-30a-5p 的机制。生物信息学和双荧光素酶报告基因检测评估了 Snai1 与 miR-31a-5p 的关系。随机将 HG 诱导的足细胞损伤小鼠分为对照组（5.5mmol/L D-葡萄糖）、HG 组（30mmol/L D-葡萄糖）、HG+丝胶组（30mmol/L D-葡萄糖+600μg/ml 丝胶）、miR-30a-5p 抑制剂 NC 组（丝胶+30mmol/L D-葡萄糖+miR-30a-5p 抑制剂阴性对照）和 miR-30a-5p 抑制剂组（丝胶+30mmol/L D-葡萄糖+miR-30a-5p 抑制剂）。通过 Transwell 检测足细胞的迁移能力。采用 RT-qPCR 和 Western blot 检测 Snai1、足细胞蛋白（podocin）、E-钙黏蛋白（E-cadherin）、纤维母细胞特异性蛋白-1（FSP-1）、紧密连接蛋白-1（ZO-1）、α-平滑肌肌动蛋白（α-SMA）、结蛋白（Desmin）和 miR-30a-5p 的表达。Snai1 是 miR-30a-5p 的一个直接靶标。与对照组和 HG+丝胶组相比，HG 组迁移的足细胞数量明显更多，Snai1、FSP-1、α-SMA 和 Desmin 水平明显更高，而 podocin、ZO-1 和 E-cadherin 水平明显更低。miR-30a-5p 抑制剂逆转了丝胶的这些作用，表现为足细胞迁移增加，Snai1、α-SMA、FSP-1 和 Desmin 的表达增加，而 podocin、ZO-1 和 E-cadherin 的表达减少。丝胶可能通过上调上皮表型标志物、下调间充质表型标志物和减少足细胞迁移来保护足细胞免受 HG 引起的损伤。其机制可能是通过靶向 miR-30a-5p 及其靶标 Snai1。
