Department of Nephrology, Sir Run Run Shaw Hospital, College of Medicine, Zhejiang University, Hangzhou, China.
Department of Thoracic Surgery, Affiliated Hangzhou First People's Hospital, School of Medicine, Westlake University, Hangzhou, China.
Immun Inflamm Dis. 2024 Nov;12(11):e70077. doi: 10.1002/iid3.70077.
Periostin mediates inflammation and fibrosis by regulating extracellular matrix adhesion, migration, and differentiation in multiple organ diseases. Studies have shown periostin mainly located in the dilated mesangium, tubulointerstitial and fibrotic regions of the diabetic kidney disease, which was negatively correlated with renal function. However, the underlying mechanism remains poorly explored.
The expression of periostin in HK-2 cells was investigated under high glucose and high concentration of TGF-β1. The signaling pathway of periostin involved in epithelial-mesenchymal transdifferentiation of HK-2 cells was also validated. The expression of periostin were investigated by RT-PCR, western blot analysis and immunofluorescence assays with different concentrations of glucose and TGF-β1. The expression of E-Cad, α-SMA and p38 proteins were also detected. The effects of periostin, E-Cad, and α-SMA in high glucose were investigated by p38 inhibitors. To demonstrate the interaction among periostin, p38 and EMT markers, periostin under high glucose and high TGF-β1 was knocked down, resulting p38 and phosphorylated p38 was evaluated.
The combined of high glucose (HG, 22 mmol/L) and high TGF-β1 (10 ng/mL) upregulated the expression of periostin obviously, stimulating the expression of α-SMA and p38 while inhibiting the expression of E-Cad. p38 inhibitors reduced the expression of periostin and α-SMA while promoted E-Cad protein expression in HK-2 cells under HG conditions. Additionally, p38-MAPK signal pathway was involved in epithelial-mesenchymal transition of human renal tubules in high glucose environment. Significant, knockdown periostin expression effectively inhibited the expression of p38 and phosphorylated p38 under the combination of HG and high TGF-β1, verifying the interaction of periostin with the p38-MAPK signaling pathway.
Periostin, a downstream factor of TGF-β1, is positively regulated by TGF-β1 under HG condition, affecting the epithelial-interstitial differentiation of HK-2 cells via p38-MAPK signaling pathway. Therefore, periostin may serve as a biomarker of renal fibrosis in diabetic kidney disease.
在多种器官疾病中，骨膜蛋白通过调节细胞外基质黏附、迁移和分化来介导炎症和纤维化。研究表明，骨膜蛋白主要位于糖尿病肾病的扩张系膜、小管间质和纤维化区域，与肾功能呈负相关。然而，其潜在机制仍未得到充分探索。
在高糖和高浓度 TGF-β1 条件下，研究了骨膜蛋白在 HK-2 细胞中的表达。验证了骨膜蛋白参与 HK-2 细胞上皮间质转化的信号通路。用不同浓度的葡萄糖和 TGF-β1 进行 RT-PCR、western blot 分析和免疫荧光检测，检测骨膜蛋白的表达。还检测了 E-Cad、α-SMA 和 p38 蛋白的表达。用 p38 抑制剂研究了骨膜蛋白、E-Cad 和 α-SMA 在高糖中的作用。为了证明骨膜蛋白、p38 和 EMT 标志物之间的相互作用，在高糖和高 TGF-β1 条件下敲低骨膜蛋白，评估 p38 和磷酸化 p38。
高糖（HG，22mmol/L）和高 TGF-β1（10ng/mL）联合明显上调骨膜蛋白的表达，刺激 α-SMA 和 p38 的表达，同时抑制 E-Cad 的表达。p38 抑制剂可降低 HG 条件下 HK-2 细胞中骨膜蛋白和 α-SMA 的表达，同时促进 E-Cad 蛋白的表达。此外，p38-MAPK 信号通路参与高糖环境下人肾小管上皮间质转化。重要的是，在 HG 和高 TGF-β1 联合作用下，敲低骨膜蛋白表达可有效抑制 p38 和磷酸化 p38 的表达，验证了骨膜蛋白与 p38-MAPK 信号通路的相互作用。
骨膜蛋白是 TGF-β1 的下游因子，在 HG 条件下受 TGF-β1 正向调节，通过 p38-MAPK 信号通路影响 HK-2 细胞的上皮间质分化。因此，骨膜蛋白可能成为糖尿病肾病中肾纤维化的生物标志物。
