Dong Fuxing, Zheng Luli, Hong Fuyuan
Department of Nephrology, Fujian Provincial Hospital, Shengli Clinical Medical College of Fujian Medical University, Fuzhou University Affiliated Provincial Hospital, Fuzhou, China.
Department of Blood Purification, Fujian Provincial Hospital, Shengli Clinical Medical College of Fujian Medical University, Fuzhou University Affiliated Provincial Hospital, Fuzhou, China.
In Vitro Cell Dev Biol Anim. 2025 Aug 28. doi: 10.1007/s11626-025-01107-1.
The mesothelial-mesenchymal transition (MMT) of peritoneal mesothelial cells is a critical factor contributing to the progression of peritoneal fibrosis. This study aimed to explore the effect of cGAS-STING signaling pathway on the MMT process in peritoneal mesothelial cells. The expressions of cGAS, STING, α-SMA, and Vimentin in HMrSV5 cells treated with high glucose were analyzed using WB. Subsequently, si-cGAS and the cGAS inhibitor RU.521 were employed to intervene in HMrSV5 cells. qPCR was utilized to evaluate the expression levels of genes involved in the cGAS-STING signaling pathway (cGAS, STING, IRF3, TBK1) and MMT-related genes (E-cadherin, Vimentin, α-SMA, TGF-β1). The protein expressions of the cGAS-STING signaling pathway and MMT-related proteins were detected by WB. The invasive capacity of cells in each cell was assessed using a Transwell assay, and the levels of pro-inflammatory cytokines (IL-6, TNF-α) in the supernatants of each cell were measured by ELISA. In the present study, we found that the expressions of cGAS, p-STING/STING, p-IRF3/IRF3, and p-TBK1/TBK1 proteins were significantly upregulated in HG-treated HMrSV5 cells. Furthermore, the activation of the cGAS-STING signaling pathway could be effectively suppressed in HMrSV5 cells transfected with si-cGAS or treated with RU.521. Additionally, treatment with si-cGAS or RU.521 not only attenuated the invasive capacity of HMrSV5 cells but also decreased the levels of pro-inflammatory cytokines and inhibited the expression of MMT-related markers. Suppression of the cGAS-STING signaling pathway mitigates HG-induced MMT in the human peritoneal mesothelial cell line HMrSV5.
腹膜间皮细胞的间皮-间充质转化（MMT）是促成腹膜纤维化进展的关键因素。本研究旨在探究cGAS-STING信号通路对腹膜间皮细胞MMT过程的影响。使用蛋白质免疫印迹法（WB）分析高糖处理的HMrSV5细胞中cGAS、STING、α-平滑肌肌动蛋白（α-SMA）和波形蛋白的表达。随后，采用小干扰RNA（si-cGAS）和cGAS抑制剂RU.521干预HMrSV5细胞。运用实时定量聚合酶链反应（qPCR）评估cGAS-STING信号通路相关基因（cGAS、STING、干扰素调节因子3（IRF3）、TANK结合激酶1（TBK1））和MMT相关基因（E-钙黏蛋白、波形蛋白、α-SMA、转化生长因子-β1（TGF-β1））的表达水平。通过WB检测cGAS-STING信号通路相关蛋白和MMT相关蛋白的表达。采用Transwell实验评估各细胞的侵袭能力，并通过酶联免疫吸附测定（ELISA）检测各细胞上清液中促炎细胞因子（白细胞介素-6（IL-6）、肿瘤坏死因子-α（TNF-α））的水平。在本研究中，我们发现高糖处理的HMrSV5细胞中cGAS、磷酸化STING/STING、磷酸化IRF3/IRF3和磷酸化TBK1/TBK1蛋白的表达显著上调。此外，在转染si-cGAS或用RU.521处理的HMrSV5细胞中，cGAS-STING信号通路的激活可被有效抑制。此外，用si-cGAS或RU.521处理不仅减弱了HMrSV5细胞的侵袭能力，还降低了促炎细胞因子水平并抑制了MMT相关标志物的表达。抑制cGAS-STING信号通路可减轻高糖诱导的人腹膜间皮细胞系HMrSV5中的MMT。
