Research Paper Volume 13, Issue 10 pp 13807—13821

Acetyl oxygen benzoate engeletin ester promotes KLF4 degradation leading to the attenuation of pulmonary fibrosis via inhibiting TGFβ1–smad/p38MAPK–lnc865/lnc556–miR-29b-2-5p–STAT3 signal pathway

Inhibition of AOBEE on lnc865 and lnc556. (A) RNA sequencing demonstrated the differently expressed RNA transcriptions in BLM group compared with those in sham group (left), and in AOBEE + BLM group compared with those in BLM group (right). Lnc865 and lnc556 upregulated in BLM group compared with those in sham group, and downregulated in AOBEE + BLM group compared with those in BLM group. (B) qRT-PCR confirmed that BLM caused a marked increase of lnc865 and lnc556 expression, whereas AOBEE caused a marked decrease of their expression in BLM-treated mice and TGFβ1-induced L929 cells. (C) The efficacy of knockdown with si-lnc865 and si-lnc556 was examined through qRT-PCR. (D) The real-time cell proliferation and migration experiments indicated that si-lnc865 and si-lnc556 inhibited the proliferation and migration of cells activated by TGFβ1. (E) Images automatically monitored by an IncuCyte S3 instrument showed that si-lnc865 and si-lnc556 repressed the migration of L929 cells activated by TGFβ1 at different time points. (F) Expression levels of collagen, vimentin, and α-SMA were downregulated in the si-lnc865 and si-lnc556 groups compared with the TGFβ1-treated group. (G) The results of rescue experiment showed that AOBEE treatment reduced the expression of α-SMA, vimentin. Overexpression of lnc865 and lnc556 reversed this treatment effect. Each bar represents the mean ± SD, n = 6; *p

Figure 4. Inhibition of AOBEE on lnc865 and lnc556. (A) RNA sequencing demonstrated the differently expressed RNA transcriptions in BLM group compared with those in sham group (left), and in AOBEE + BLM group compared with those in BLM group (right). Lnc865 and lnc556 upregulated in BLM group compared with those in sham group, and downregulated in AOBEE + BLM group compared with those in BLM group. (B) qRT-PCR confirmed that BLM caused a marked increase of lnc865 and lnc556 expression, whereas AOBEE caused a marked decrease of their expression in BLM-treated mice and TGFβ1-induced L929 cells. (C) The efficacy of knockdown with si-lnc865 and si-lnc556 was examined through qRT-PCR. (D) The real-time cell proliferation and migration experiments indicated that si-lnc865 and si-lnc556 inhibited the proliferation and migration of cells activated by TGFβ1. (E) Images automatically monitored by an IncuCyte S3 instrument showed that si-lnc865 and si-lnc556 repressed the migration of L929 cells activated by TGFβ1 at different time points. (F) Expression levels of collagen, vimentin, and α-SMA were downregulated in the si-lnc865 and si-lnc556 groups compared with the TGFβ1-treated group. (G) The results of rescue experiment showed that AOBEE treatment reduced the expression of α-SMA, vimentin. Overexpression of lnc865 and lnc556 reversed this treatment effect. Each bar represents the mean ± SD, n = 6; *p < 0.05, **p < 0.01.