Epidermal growth factor regulation by autophagy-mediated lncRNA H19 in murine intestinal tract after severe burn
Epidermal growth factor regulation by autophagy-mediated lncRNA H19 in murine intestinal tract after severe burn
复制标题
严重烧伤后自噬介导的lncRNA H19对小鼠肠道表皮生长因子的调节
DOI：
10.1111/jcmm.15262
复制
发表时间：
2020-05-01
影响因子：
5.3
通讯作者：
Sun, Yong
中科院分区：
文献类型：
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作者：
Li, Cuijie;Zhuang, Mengmeng;Sun, Yong
探讨自噬介导的长链非编码RNA（lncRNA）H19对严重烧伤小鼠肠道表皮生长因子（EGF）表达的调节作用。C57 BL/6 J小鼠接受三度烧伤至总体表面积的30%。使用雷帕霉素和3-甲基腺嘌呤（3-MA）激活和抑制自噬，并通过Western印迹法评估LC 3和Beclin 1水平的变化。通过qRT-PCR检测自噬对lncRNA H19的影响。采用腺病毒介导的lncRNA H19在IEC-6细胞中的过表达，通过生物信息学分析、Western印迹和qRT-PCR评估lncRNA H19对EGF和let-7 g的影响。使用let-7 g模拟物/抑制剂过表达/抑制let-7 g，并使用qRT-PCR和Western印迹检测let-7 g对EGF的影响。雷帕霉素处理后肠组织和IEC-6细胞中LC 3-II、Beclin 1和lncRNA H19的表达水平增加，而3-MA处理后则逆转。烧伤后肠组织中LC 3-II、Beclin 1和lncRNA H19水平升高，雷帕霉素处理后升高更明显，而3-MA处理后降低。在IEC-6细胞中lncRNA H19过表达分别导致EGF和let-7 g表达水平的增加和降低。此外，let-7 g的过表达和抑制分别导致EGF表达的减少和增加。综上所述，严重烧伤后肠道自噬被激活，这可以增加lncRNA H19的转录水平。lncRNA H19可能通过let-7 g调节烧伤后肠粘膜损伤后EGF的修复。
To investigate the regulation of epidermal growth factor (EGF) by autophagy-mediated long non-coding RNA (lncRNA) H19 in the intestinal tracts of severely burned mice. C57BL/6J mice received third-degree burns to 30% of the total body surface area. Rapamycin and 3-methyladenine (3-MA) were used to activate and inhibit autophagy, and the changes in LC3 and Beclin1 levels were assessed by Western blotting. The effect of autophagy on lncRNA H19 was detected by qRT-PCR. Adenovirus-mediated overexpression of lncRNA H19 in IEC-6 cells was used to assess the effects of lncRNA H19 on EGF and let-7g via bioinformatics analysis, Western blotting and qRT-PCR. let-7g mimic/inhibitor was used to overexpress/inhibit let-7g, and qRT-PCR and Western blotting were used to detect the effects of let-7g on EGF. The expression levels of LC3-II, Beclin1 and lncRNA H19 were increased in intestinal tissues and IEC-6 cells after rapamycin treatment but were reversed after 3-MA treatment. LC3-II, Beclin1 and lncRNA H19 levels increased in intestinal tissues after the burn, and these increases were more significant after rapamycin treatment but decreased after 3-MA treatment. The lncRNA H19 overexpression in IEC-6 cells resulted in increased and decreased expression levels of EGF and let-7g, respectively. Furthermore, overexpression and inhibition of let-7g resulted in decreased and increased expression of EGF, respectively. Taken together, intestinal autophagy is activated after a serious burn, which can increase the transcription level of lncRNA H19. lncRNA H19 may regulate the repair of EGF via let-7g following intestinal mucosa injury after a burn.