Jiangsu Key Laboratory of New Drug Research and Clinical Pharmacy, Xuzhou Medical University, China.
Jiangsu Key Laboratory of New Drug Research and Clinical Pharmacy, Xuzhou Medical University, China; State Key Laboratory of Bioactive Substance and Function of Natural Medicines, Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College, China.
Metabolism. 2023 Jul;144:155376. doi: 10.1016/j.metabol.2022.155376. Epub 2022 Dec 12.
Renal interstitial fibrosis (RIF) is one of the main features of diabetic nephropathy (DN), but the molecular mechanisms mediating RIF in DN has yet been fully understood. S100A8 and S100A9 are the proteins associated with immune and inflammation response. Here we reported the expression of S100A8 and S100A9 were significantly increased on tubular epithelial cells in diabetic kidneys through a proteomic analysis.
We detected the expression of S100A8/A9 in diabetic kidneys by using immunoblotting, real-time PCR and immunostaining. RNA silencing and overexpression were performed by using S100A8/A9 expression/knockdown lentivirus to investigate the connection between S100A8/A9 and epithelial to mesenchymal transition (EMT) process. We also identify the expression of TLR4/NFκB pathway-related molecules in the case mentioned above. Afterwards a CO-IP assay was used to verify that compound AB38b ameliorates the EMT by interfering S100A8/A9 expression.
The expression of S100A8 and S100A9 were significantly increased on tubular epithelial cells in diabetic kidneys. S100A8/A9 knocking-down alleviate and over-expression promote the renal interstitial fibrosis of diabetic mice. Mechanically, high levels of S100A8/A9 expression in tubular epithelial cells during diabetic condition activated the TLR4/NF-κB signal pathway which promoted the EMT process and finally led to RIF progression. S100A8/A9 knockdown ameliorated RIF of diabetic mice. Further experiments revealed that compound AB38b inhibited the EMT progression of tubular epithelial cells induced by S100A8/A9 through interfering the expressions of S100A8/A9.
Our study suggest that abnormal expression of S100A8/A9 in the disease condition promotes EMT process and RIF through TLR4/NF-κB signal pathway. Using small molecular inhibitor AB38b to inhibit the abnormal expressions of S100A8/A9 might be a novel therapeutic strategy in treating DN.
肾间质纤维化(RIF)是糖尿病肾病(DN)的主要特征之一,但介导 DN 中 RIF 的分子机制尚未完全阐明。S100A8 和 S100A9 是与免疫和炎症反应相关的蛋白质。在这里,我们通过蛋白质组学分析报道了在糖尿病肾脏中肾小管上皮细胞中 S100A8 和 S100A9 的表达显著增加。
我们通过免疫印迹、实时 PCR 和免疫染色检测糖尿病肾脏中 S100A8/A9 的表达。通过 S100A8/A9 表达/敲低慢病毒转染进行 RNA 沉默和过表达,以研究 S100A8/A9 与上皮间质转化(EMT)过程之间的联系。我们还鉴定了上述情况下 TLR4/NFκB 途径相关分子的表达。随后进行 CO-IP 测定以验证化合物 AB38b 通过干扰 S100A8/A9 表达来改善 EMT。
在糖尿病肾脏中,肾小管上皮细胞中 S100A8 和 S100A9 的表达显著增加。S100A8/A9 敲低减轻和过表达促进糖尿病小鼠的肾间质纤维化。从机制上讲,在糖尿病条件下肾小管上皮细胞中高表达的 S100A8/A9 激活了 TLR4/NF-κB 信号通路,促进了 EMT 过程,最终导致 RIF 进展。S100A8/A9 敲低减轻了糖尿病小鼠的 RIF。进一步的实验表明,化合物 AB38b 通过干扰 S100A8/A9 的表达,抑制 S100A8/A9 诱导的肾小管上皮细胞 EMT 进展。
我们的研究表明,疾病状态下 S100A8/A9 的异常表达通过 TLR4/NF-κB 信号通路促进 EMT 过程和 RIF。使用小分子抑制剂 AB38b 抑制 S100A8/A9 的异常表达可能是治疗 DN 的一种新的治疗策略。