肠上皮Depdc5/mTORC1信号轴调控小鼠小肠上皮稳态

doi:10.16352/j.issn.1001-6325.2022.08.1213

基础医学与临床 ›› 2022, Vol. 42 ›› Issue (8): 1213-1219.doi: 10.16352/j.issn.1001-6325.2022.08.1213

肠上皮Depdc5/mTORC1信号轴调控小鼠小肠上皮稳态

张新格¹, 马洁², 王庆志¹, 辛悦¹, 杨晨妍¹, 熊熙文^1*

新乡医学院 1.法医学院; 2.基础医学院人体解剖与组织胚胎学系, 河南新乡 453000

收稿日期:2021-03-04 修回日期:2021-10-15 出版日期:2022-08-05 发布日期:2022-08-01
通讯作者: ^*xwxiong@xxmu.edu.cn
基金资助:
NSFC-河南联合基金(U1904132);河南省高校科技创新人才支持计划(20HASTIT046);河南省高等学校青年骨干教师培养计划(2017GGJS110);新乡市科技攻关项目(GG2019008)

Gut epithelial Depdc5/mTORC1 signaling axis regulates mouse intestinal epithelial homeostasis

ZHANG Xin-ge¹, MA Jie², WANG Qing-zhi¹, XIN Yue¹, YANG Chen-yan¹, XIONG Xi-wen^1*

1. School of Forensic Medicine; 2. Department of Human Anatomy & Histoembryology, School of Basic Medical Sciences, Xinxiang Medical University, Xinxiang 453000, China

Received:2021-03-04 Revised:2021-10-15 Online:2022-08-05 Published:2022-08-01
Contact: ^*xwxiong@xxmu.edu.cn

摘要/Abstract

摘要： 目的探讨Depdc5敲除介导的mTORC1信号通路持续激活对小鼠肠道上皮稳态的调控作用。方法将小鼠分为4组,Depdc5^flox/flox组(对照组)、Depdc5^flox/flox:Villin-Cre组(IKO组)、rapamycin-Depdc5^flox/flox组(rap-Ctrl组)、rapamycin-Depdc5^flox/flox:Villin-Cre组(rap-IKO组),每组各5只。Western blot检测肠道上皮DEPDC5蛋白表达及mTORC1下游分子S6的磷酸化(PS6)水平。RT-qPCR检测小肠组织中不同类型细胞标记基因Dclk1、Trpm5、Muc2 mRNA的表达。苏木精-伊红染色(HE染色)对组织形态学进行检查。免疫组化染色(IHC)检测簇细胞、潘氏细胞、增殖细胞的数目。阿尔新蓝染色法检测杯状细胞的数目。结果 IKO组小鼠小肠和结肠组织中PS6蛋白表达为0.957±0.028高于对照组的0.598±0.041(P＜0.05),并且IKO组小鼠小肠上皮不同类型细胞标记基因Dclk1、Trpm5、Muc2的mRNA均低于对照组(P＜0.05)。此外,IKO组小鼠小肠中簇细胞、潘氏细胞、杯状细胞的细胞数目显著低于对照组,隐窝深度显著高于对照组(P＜0.05),增殖细胞显著高于对照组(P＜0.05)。结论 mTORC1过度激活抑制肠道上皮细胞(IECs)的分化,破坏小鼠肠上皮细胞稳态。

关键词: Depdc5, mTORC1, 肠上皮细胞, 雷帕霉素

Abstract: Objective To investigate the effect of over-activation of mTORC1 on mouse intestinal epithelial homeostasis by conditional knockout of Depdc5. Methods Mice were divided into Depdc5^flox/flox group (control group), Depdc5^flox/flox:Villin-Cre group (IKO group), rapamycin-Depdc5^flox/flox group (rap-Ctrl group) rapamycin-Depdc5^flox/flox:Villin-Cre group (rap-IKO group). Western blot was used to detect the protein expression of DEPDC5 and the phosphorylation S6 (PS6) which is the downstream molecule of mTORC1 in the intestinal epithelium. qPCR was applied to detect the mRNA levels of Dclk1, Trpm5 and Muc2 in intestinal epithelial cells (IECs). Hematoxylin-eosin (HE) staining was used to observe the intestinal morphology changes. Immunohistochemical (IHC) staining and alcian blue staining were used to detect the number of tuft cells, Paneth cells, proliferative cells and goblet cells. Results The protein expression of PS6 in the small intestine and colon from the mice of IKO group was obviously up-regulated than that of the control group (0.957±0.028 vs. 0.598±0.041) (P＜0.05). The mRNA levels of Dclk1, Trpm5, and Muc2 were significantly decreased in the IKO group than those in the control group (P＜0.05). The numbers of tuft cells, Paneth cells and goblet cells in the IKO group were all significantly reduced than that in the control group. Nevertheless, the crypt depth in the IKO group was significantly higher than that of the control group (P＜0.05), and the number of proliferative cells was also increased in the IKO group(P＜0.05). Conclusions The over-activation of mTORC1 inhibits the differentiation of IECs and impairs the homeostasis of intestinal epithelial.

Key words: Depdc5, mTORC1, IECs, rapamycin

中图分类号:

R574.62

张新格, 马洁, 王庆志, 辛悦, 杨晨妍, 熊熙文. 肠上皮Depdc5/mTORC1信号轴调控小鼠小肠上皮稳态[J]. 基础医学与临床, 2022, 42(8): 1213-1219.

ZHANG Xin-ge, MA Jie, WANG Qing-zhi, XIN Yue, YANG Chen-yan, XIONG Xi-wen. Gut epithelial Depdc5/mTORC1 signaling axis regulates mouse intestinal epithelial homeostasis[J]. Basic & Clinical Medicine, 2022, 42(8): 1213-1219.

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肠上皮Depdc5/mTORC1信号轴调控小鼠小肠上皮稳态

Gut epithelial Depdc5/mTORC1 signaling axis regulates mouse intestinal epithelial homeostasis

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