Department of Critical Care Medicine, Beijing Friendship Hospital Affiliated to Capital Medical University, Beijing, China.
J Int Med Res. 2021 May;49(5):3000605211012665. doi: 10.1177/03000605211012665.
To explore the effects of wedelolactone (WEL) on sepsis-induced renal injury in the human renal proximal tubular epithelial cell line HK-2.
HK-2 cells were stimulated by 1 µg/ml lipopolysaccharide (LPS) to trigger renal injury . HK-2 cells were pretreated with or without WEL (0.1, 1 and 10 µM) before LPS stimulation. Protein and mRNA analyses were performed using enzyme-linked immunosorbent assays, Western blot analysis and quantitative reverse transcription-polymerase chain reaction. The MTT assay and flow cytometry were used to measure cell viability and the rate of cell apoptosis. Protein tyrosine phosphatase non-receptor type 2 (PTPN2) knockdown was induced by the transection of HK-2 cells with short hairpin RNA.
Cell viability was significantly increased in a dose-dependent manner by WEL in LPS-induced HK-2 cells. WEL also decreased the levels of four inflammatory cytokines and cell apoptosis in LPS-induced HK-2 cells. The level of PTPN2 was increased after WEL treatment. PTPN2 knockdown partly abolished the inhibitory effects of WEL on cell apoptosis, the levels of inflammatory cytokines and on p38 mitogen-activated protein kinase/nuclear factor-kappaB signalling in LPS-induced HK-2 cells.
WEL improved renal injury by suppressing inflammation and cell apoptosis through upregulating PTPN2 in HK-2 cells. PTPN2 might be used as a potential therapeutic target for LPS-induced sepsis.
探讨冬凌草甲素（WEL）对脂多糖（LPS）诱导的人近端肾小管上皮细胞系 HK-2 细胞肾损伤的影响。
用 1μg/ml LPS 刺激 HK-2 细胞诱发肾损伤，用 WEL（0.1、1 和 10μM）预处理 LPS 刺激前的 HK-2 细胞。用酶联免疫吸附试验、Western blot 分析和定量逆转录聚合酶链反应进行蛋白和 mRNA 分析。MTT 测定和流式细胞术分别用于测定细胞活力和细胞凋亡率。通过转染短发夹 RNA 使 HK-2 细胞中的蛋白酪氨酸磷酸酶非受体型 2（PTPN2）敲低。
WEL 以剂量依赖性方式显著增加 LPS 诱导的 HK-2 细胞中的细胞活力。WEL 还降低了 LPS 诱导的 HK-2 细胞中四种炎症细胞因子和细胞凋亡的水平。WEL 处理后 PTPN2 水平增加。PTPN2 敲低部分消除了 WEL 对 LPS 诱导的 HK-2 细胞中细胞凋亡、炎症细胞因子水平和 p38 丝裂原活化蛋白激酶/核因子-κB 信号的抑制作用。
WEL 通过在上调 LPS 诱导的 HK-2 细胞中的 PTPN2 来抑制炎症和细胞凋亡，从而改善肾损伤。PTPN2 可能成为 LPS 诱导的脓毒症的潜在治疗靶点。
