Effects of DNMT3A regulating Drp1 on proliferation and migration of activated hepatic stellate cells / 中国药理学通报

ABSTRACT

Aim To investigate the effects of DNMT3A regulating Drp1 mediated mitochondrial fission on the proliferation and migration of active hepatic stellate cells. Methods HSC-T6 cells were activated by 5 μg·L-1 TGF-β1 for 24 h, and DNMT3A lentivirus infection model was established to silence DNMT3A. The experiment was divided into control group, TGF-β1 group, TGF-β1+LV5-NC group and TGF-β1+ LV5-DNMT3A group. The effects of DNMT3A on related mRNA and protein expression were detected by RT-qPCR and Western blot. The cell proliferation was detected by CCK-8. The effect of DNMT3A on the migration ability of HSCs cells was observed by Wound healing assay and Transwell migration assay. Results Lentivirus infection successfully constructed a DNMT3A silencing model. Compared with the control group, the level of DNMT3A significantly increased, the mRNA and protein levels of the fibrosis markers collagen and α-SMA in the TGF-β1 group significantly increased, and the mRNA and protein levels of the mitochondrial fission marker Drp1 significantly increased; At the same time, the proliferation and migration ability of HSCs cells was significantly improved. Compared with the NC group, the DNMT3A level of the DNMT3A silenced group was significantly reduced, the expressions of collagen I, α-SMA and Drp1 were significantly inhibited, and the proliferation and migration capabilities of HSCs were also significantly inhibited. Conclusions Silencing DNMT3A inhibits the level of Drp1 and inhibits the proliferation and migration of HSCs at the same time. It is suggested suggest that DNMT3A-mediated low level DNA methylation modification may inhibit the occurrence of mitochondrial fission by inhibiting the level of Drp1, thereby inhibiting the activation of HSCs and affecting the occurrence and development of liver fibrosis. ,,,,.