PERK is essential for proliferation of intestinal stem cells in mice.
PERK is essential for proliferation of intestinal stem cells in mice.
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DOI：
10.1016/j.yexcr.2018.12.009
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发表时间：
2019-02
影响因子：
3.7
通讯作者：
Zhihao Liu;Jinli Liao;Hongyan Wei;Zhen Yang;Jingjing Liu;Jia Xu;Xinlin Wu;H. Zhan
中科院分区：
文献类型：
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作者：
Zhihao Liu;Jinli Liao;Hongyan Wei;Zhen Yang;Jingjing Liu;Jia Xu;Xinlin Wu;H. Zhan
蛋白激酶RNA样内质网激酶（PERK）是一种内质网应激传感器，具有促生存能力，有助于细胞稳态和存活。富含亮氨酸重复序列的G蛋白偶联受体5（Lgr 5）已被认为是肠上皮细胞中的干细胞标志物。为了确定PERK是否调节肠道干细胞的增殖，我们研究了PERK敲低对小鼠肠道Lgr 5阳性干细胞的影响。Lgr 5-EGFP敲入小鼠每天两次喂食慢病毒-PERK shRNA，持续3天。用慢病毒-PERK shRNA处理分离的肠Lgr 5阳性干细胞。在体内、体外以及两种肠上皮损伤模型中检测肠干细胞中Lgr 5阳性细胞的数量、增殖和凋亡指数、增殖和分化的几种生物标志物以及Akt的表达。PERK基因敲低可显著减少Lgr 5阳性细胞的数量和增殖，诱导Lgr 5阳性细胞中多种增殖标志物的低表达和多种分化标志物的高表达，促进Lgr 5阳性细胞的凋亡，并降低肠Lgr 5阳性干细胞中Akt的表达。在放疗和脓毒症引起的肠损伤中也观察到类似的结果。此外，PERK抑制显著降低了小鼠对辐射和脓毒症的存活率。这些结果表明PERK在小鼠肠干细胞的增殖和存活中起关键作用。
Protein kinase RNA-like Endoplasmic Reticulum Kinase (PERK) is an endoplasmic reticulum stress sensor that possesses pro-survival capability and contributes to cell homeostasis and survival. Leucine-rich repeat-containing G-protein coupled receptor 5 (Lgr5) has been recognized as a stem cell marker in intestinal epithelial cells. To determine whether PERK modulates the proliferation of intestinal stem cells, we investigated the effects of PERK knock-down on intestinal Lgr5-positive stem cells in mice.Lgr5-EGFPknock-in mice were fed with lentivirus-PERK shRNA twice a day for three days. Isolated intestinal Lgr5-positive stem cells were treated with lentivirus-PERK shRNA. The number of Lgr5-positive cells, the proliferation and apoptotic indices, several biomarkers for proliferation and differentiation, and Akt expression in intestinal stem cells were detected in vivo, in vitro and in two intestinal epithelial injury models caused by radiotherapy and sepsis. PERK knock-down could significantly diminish the number and proliferation of Lgr5-positive cells, induce the low expression of several proliferation markers and the high expression of several differentiation markers in Lgr5-positive cells, enhance the apoptotic Lgr5-positive cells, and reduce the Akt expression in intestinal Lgr5-positive stem cells. Similar results were observed in radiotherapy- and sepsis-induced intestinal injuries. Moreover, PERK inhibition markedly decreased the survival of mice in response to radiation and sepsis. These results suggest a critical role for PERK in the proliferation and survival of intestinal stem cells in mice.