Research Paper Volume 15, Issue 12 pp 5514—5534

Figure 6. Rescue experiments proved that hucMSCs treatment alleviated pulmonary fibrosis by regulating the upstream circANKRD42-miR-136-5p of YAP1 in mice. (A) Schematic of the administration of overexpressed circANKRD42, circANKRD42 mutation, BLM, or hucMSCs into mice. (B) qRT-PCR revealed that circANKRD42 was substantially upregulated in overexpressed circANKRD42 group compared with other groups and reversed the inhibitory effect of hucMSCs treatment. The effect of circANKRD42 mutation was weaker than that of overexpressed circANKRD42. (C) circANKRD42 overexpression reversed the upward trend of miR-136-5p caused by hucMSCs treatment, and the effect of circANKRD42 mutation on miR-136-5p was weaker than that of overexpressed circANKRD42. (D) MicroCT imaging system for small animal showed that circANKRD42 overexpression reversed the therapeutic effect of hucMSCs and aggravated the degree of fibrosis. The effect of circANKRD42 mutation on fibrosis was weaker than that of overexpressed circANKRD42. (E) H&E and Masson staining exhibited that BLM caused excessive collagen deposition, bulk fibroblast paraplasm displacing alveolar space in lung tissues, and near disappearance of alveoli pulmonis. Fibrosis was improved in the BLM+hucMSCs group. Overexpressed circANKRD42 reversed the therapeutic effect of hucMSCs, but the effect of circANKRD42 mutation was weaker than that of overexpressed circANKRD42. (F) BLM deteriorated the lung function and body weight. Overexpressed circANKRD42 reversed the ameliorating effect of hucMSCs on the lung function and body weight. The effect of circANKRD42 mutation on the lung function and body weight was weaker than that of overexpressed circANKRD42. (G) Western blot identified that BLM promoted the expression levels of fibrotic and differentiation proteins. Overexpressed circANKRD42 reversed the effect of hucMSCs on these proteins. The effect of circANKRD42 mutation was weaker than that of overexpressed circANKRD42. Each bar represents the mean ± SD; n = 6; ^*p < 0.05.