Department of Burn Surgery, The Affiliated Huaihai Hospital of Xuzhou Medical University, Xuzhou, China.
Department of Burn Surgery, The 71st Group Army Hospital of PLA, Xuzhou, China.
J Cell Mol Med. 2020 May;24(10):5878-5887. doi: 10.1111/jcmm.15262. Epub 2020 Apr 16.
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.
研究自噬通过长链非编码 RNA（lncRNA）H19 对严重烧伤小鼠肠道中表皮生长因子（EGF）的调节作用。将 C57BL/6J 小鼠的 30%的体表面积造成三度烧伤。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 blot 评估 LC3 和 Beclin1 水平的变化。通过 qRT-PCR 检测自噬对 lncRNA H19 的影响。使用腺病毒介导的 lncRNA H19 在 IEC-6 细胞中的过表达来评估通过生物信息学分析、Western blot 和 qRT-PCR 检测 lncRNA H19 对 EGF 和 let-7g 的影响。使用 let-7g 模拟物/抑制剂过表达/抑制 let-7g，并通过 qRT-PCR 和 Western blot 检测 let-7g 对 EGF 的影响。雷帕霉素处理后，肠组织和 IEC-6 细胞中 LC3-II、Beclin1 和 lncRNA H19 的表达水平增加，但 3-MA 处理后逆转。烧伤后肠组织中 LC3-II、Beclin1 和 lncRNA H19 的水平增加，雷帕霉素处理后增加更为明显，但 3-MA 处理后减少。IEC-6 细胞中 lncRNA H19 的过表达导致 EGF 和 let-7g 的表达水平分别增加和减少。此外，let-7g 的过表达和抑制分别导致 EGF 的表达减少和增加。总之，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 可能通过烧伤后肠黏膜损伤时的 let-7g 调节 EGF 的修复。
