Luo X, Zhang L, Han G-D, Lu P, Zhang Y
Department of Pharmacology, Cangzhou Central Hospital, Cangzhou, China.
Department of Clinical Laboratory, Cangzhou Central Hospital, Cangzhou, China.
Hum Exp Toxicol. 2021 Feb;40(2):369-379. doi: 10.1177/0960327120952158. Epub 2020 Aug 28.
To explore the potential function of MDM2-mediated Notch/hes1 signaling pathway in cisplatin-induced renal injury.
The acute renal injury models of mice after intraperitoneal injection of cisplatin , and the apoptotic models of human renal tubular epithelial (HK-2) cells induced by cisplatin in , were conducted respectively. The renal function-related parameters were measured. The renal tissue pathological changes and apoptosis were observed by PAS staining and TUNEL staining, respectively. Cell viability and apoptosis were detected by MTT and flow cytometry. Notch/hes1 pathway-related proteins were tested by Western blotting.
After mice injected by cisplatin, the levels of Cr, BUN, urine cystatin C, urine NGAL and urine ACR were increased and GFR was decreased with the elevation of renal tubular injury scores, the upregulation of the expressions of MDM2, N1ICD, Hes1 and Cleaved caspase-3, as well as the enhancement of cell apoptosis accompanying decreased ratio of Bcl-2/Bax. However, these cisplatin-induced renal injuries of mice have been improved by MDM2 inhibition. Besides, the declined viability, increased cytotoxicity, and enhanced apoptosis were observed in cisplatin-induced HK-2 cells, with the activated Notch/hes1 pathway. Notably, the phenomenon was alleviated in cisplatin-induced HK-2 cells transfected with MDM2 shRNA, but was severer in those co-treated with AdMDM2. Moreover, Notch1 siRNA can reverse the injury of AdMDM2 on HK-2 cells.
Inhibiting MDM2 could reduce cell apoptosis through blocking Notch/hes1 signaling pathway, thus alleviating the acute renal injury caused by cisplatin.
探讨MDM2介导的Notch/hes1信号通路在顺铂诱导的肾损伤中的潜在作用。
分别构建腹腔注射顺铂后的小鼠急性肾损伤模型以及顺铂诱导的人肾小管上皮（HK-2）细胞凋亡模型。检测肾功能相关参数。分别通过PAS染色和TUNEL染色观察肾组织病理变化和细胞凋亡情况。采用MTT法和流式细胞术检测细胞活力和凋亡情况。通过蛋白质免疫印迹法检测Notch/hes1通路相关蛋白。
顺铂注射后的小鼠，随着肾小管损伤评分升高、MDM2、N1ICD、Hes1和Cleaved caspase-3表达上调、细胞凋亡增强以及Bcl-2/Bax比值降低，Cr、BUN、尿胱抑素C、尿中性粒细胞明胶酶相关脂质运载蛋白和尿ACR水平升高，肾小球滤过率降低。然而，抑制MDM2可改善顺铂诱导的小鼠肾损伤。此外，在顺铂诱导的HK-2细胞中观察到活力下降、细胞毒性增加和凋亡增强，同时Notch/hes1通路激活。值得注意的是，在转染MDM2 shRNA的顺铂诱导的HK-2细胞中该现象得到缓解，但在与AdMDM2共同处理的细胞中更严重。此外，Notch1 siRNA可逆转AdMDM2对HK-2细胞的损伤。
抑制MDM2可通过阻断Notch/hes1信号通路减少细胞凋亡，从而减轻顺铂所致的急性肾损伤。
