Department of Nephrology, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200127, China.
Department of Nephrology, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200127, China.
Life Sci. 2020 Aug 1;254:117791. doi: 10.1016/j.lfs.2020.117791. Epub 2020 May 19.
Sepsis-induced acute kidney injury (SI-AKI) is the fifth most common cause of hospital-acquired acute kidney injury. Pannexin1 (Panx1) triggers inflammation and apoptosis which act as crucial factors in the pathogenesis of SI-AKI. We aimed to investigate the expression of Panx1 and its role on the inflammation and apoptosis in SI-AKI.
SI-AKI model was established by lipopolysaccharide (LPS) injection in mice and LPS-treated HK-2 cells in vitro. Panx1 was inhibited by pretreating with carbenoxolone (CBX) or small interfering RNA in vivo and vitro, respectively. The expression of Panx1 was determined by qPCR, western blot and immunohistochemistry (IHC). Kidney damage was evaluated by kidney function, histopathological examination and AKI biomarkers. Inflammatory cytokines were detected by qPCR and ELISA. Apoptosis was detected by TUNEL staining and the expression of apoptosis-related proteins. The activation of nucleotide-binding domain-like receptor protein 3 (NLRP3) inflammasome was measured by western blot.
Panx1 increased in LPS-induced SI-AKI mice and HK-2 cells, as well as in SI-AKI patients. CBX alleviated the renal function and pathological damage, as well as decreased the mRNA of kidney injury molecule (KIM-1) and neutrophil gelatinase-associated lipocalin (NGAL). Inhibiting Panx1 decreased the production of IL-1β, IL-6 and TNF-α, as well as tubular cell apoptosis in SI-AKI. Inhibiting Panx1 suppressed inflammatory cytokines and apoptosis via inhibiting NLRP3 inflammasome activation and regulating apoptotic protein Bax and Bcl2 expression, respectively.
These observations suggest that pharmacological inhibition of Panx1 might be a potential approach in the clinical therapy of SI-AKI.
脓毒症诱导的急性肾损伤（SI-AKI）是医院获得性急性肾损伤的第五大常见原因。连接蛋白 1（Panx1）触发炎症和细胞凋亡，这些炎症和细胞凋亡是 SI-AKI 发病机制中的关键因素。我们旨在研究 Panx1 的表达及其在 SI-AKI 中的炎症和细胞凋亡中的作用。
通过 LPS 注射在小鼠中建立 SI-AKI 模型，并在体外通过 LPS 处理 HK-2 细胞。通过体内和体外分别用 carbenoxolone（CBX）或小干扰 RNA 预处理来抑制 Panx1。通过 qPCR、western blot 和免疫组织化学（IHC）测定 Panx1 的表达。通过肾功能、组织病理学检查和 AKI 生物标志物评估肾脏损伤。通过 qPCR 和 ELISA 检测炎性细胞因子。通过 TUNEL 染色和凋亡相关蛋白的表达检测细胞凋亡。通过 western blot 测量核苷酸结合域样受体蛋白 3（NLRP3）炎性小体的激活。
在 LPS 诱导的 SI-AKI 小鼠和 HK-2 细胞以及 SI-AKI 患者中，Panx1 增加。CBX 缓解了肾功能和病理损伤，并降低了肾脏损伤分子（KIM-1）和中性粒细胞明胶酶相关脂质运载蛋白（NGAL）的 mRNA。抑制 Panx1 减少了 SI-AKI 中的 IL-1β、IL-6 和 TNF-α产生以及管状细胞凋亡。抑制 Panx1 通过抑制 NLRP3 炎性小体的激活和调节凋亡蛋白 Bax 和 Bcl2 的表达来抑制炎症细胞因子和凋亡。
这些观察结果表明，药理学抑制 Panx1 可能是 SI-AKI 临床治疗的一种潜在方法。
