RIPK1抑制糖皮质激素体外诱导的间充质干细胞凋亡

RIPK1 axis inhibits glucocorticoid induced-apoptosis of mesenchymal stem cells in vitro

目的研究受体相互作用蛋白激酶1(receptor-interacting protein kinase 1,RIPK1)抑制糖皮质激素(glucocorticoid,GC)体外诱导的间充质干细胞(mesenchymal stem cell,MSC)的凋亡效果及其调控机制。方法将SD大鼠来源的MSC,分为3组:GC+MSC组,GC+RIPK1-MSC组和生理盐水+MSC组(对照组)。通过形态学观察、MTT分析、AO/EB染色、Hoechest 333342染色、Live/Dead染色、TUNEL凋亡检测及Western blot分析等方法确定抑制效果,并分析其中涉及的关键蛋白和信号通路。结果 GC诱导24h后,MSC发生凋亡,并证实RIPK1可对抗这种凋亡。RIPK1上调伴随TRAF2/cIAP1/cFLIPL水平升高,其为对抗凋亡的关键蛋白通路。结论 RIPK1可抑制GC诱导的MSC凋亡,为早期阻遏股骨头坏死(osteonecrosis of femoral head,ONFH)提供了新思路。

Objective To determining whether receptor-interacting protein kinase 1 （RIPK1） could inhibit glucocorticoid （GC） induced-apoptosis of mesenchymal stem cells （MSC） in vitro and its underlying mechanisms.Methods Experimental MSCs obtained from SD rats were divided into 3 groups：GCs ＋ MSCs group,GCs ＋ RIPK1-MSCs group and vehicle control group.TUNEL,AO/EB,Hoechst 333342 and Live/Dead staining were used to confirm the effects of RIPK1 axis inhibiting apoptosis of MSCs caused by GCs;and Western blot was used to determine the key proteins involved in this signal pathway.Results All the testing methods demonstrated the effectiveness of RIPK1 on inhibiting GC induced-apoptosis of MSCs.RIPK1 clearly has a protective role in apoptosis signaling.It up-regulates anti-apoptotic proteins,principally TRAF2/cIAP1/cFLIPL,to prevent the activation of caspase-8 and-3.Conclusions RIPK1 inhibits GC induced-apoptosis of MSCs in vitro,which provides a new possible solution for the treatment of osteonecrosis of femoral head （ONFH） at the early stage.