METHODS: C57BL/6 male mice, alveolar epithelial cell-specific FUT8 conditional knockout (CKO) mice, and MLE12 cells were administered bleomycin (BLM), FUT8 siRNA, and mouse BMSCs, respectively. Experimental techniques including tissue staining, western blotting, immunofluorescence, autophagic flux detection, and flow cytometry were utilized in this study.
RESULTS: First, we found that autophagy was inhibited while FUT8 expression was elevated in PF mice and BLM-induced AEC injury models. Subsequently, CKO mice and MLE12 cells transfected with FUT8 siRNA were employed to demonstrate that inhibition of CF modification induces autophagy in AECs and mitigates PF. Finally, mouse BMSCs were utilized to demonstrate that they alleviate the detrimental autophagy of AECs by inhibiting CF modification and decreasing PF.
CONCLUSIONS: Suppression of CF modification enhanced the suppression of AEC autophagy and reduced PF in mice. Additionally, through the prevention of CF modification, BMSCs can assist AECs deficient in autophagy and partially alleviate PF.
方法:C57BL/6雄性小鼠,肺泡上皮细胞特异性FUT8条件敲除(CKO)小鼠,和MLE12细胞给予博来霉素(BLM),FUT8siRNA,和小鼠BMSCs,分别。实验技术包括组织染色,西方印迹,免疫荧光,自噬通量检测,和流式细胞术在这项研究中使用。
结果:首先,我们发现在PF小鼠和BLM诱导的AEC损伤模型中,自噬受到抑制,而FUT8表达升高。随后,使用CKO小鼠和用FUT8siRNA转染的MLE12细胞来证明CF修饰的抑制诱导AEC中的自噬并减轻PF。最后,小鼠BMSCs被用来证明它们通过抑制CF修饰和降低PF来减轻AECs的有害自噬。
结论:抑制CF修饰可增强小鼠AEC自噬的抑制和降低PF。此外,通过防止CF修改,BMSCs可以辅助自噬缺陷的AECs,部分缓解PF。