Fig. 8

Antidepressant effect of LBRD standard decoction on the mRNA/miRNA regulatory network of GABA and BDNF expression. A The interactive networks of four overlapped miRNAs and six mRNAs in the LBRD standard decoction treated group from RNA sequencing data of CUMS-induced animal and CORT-induced cell depression model. Green symbols denote miRNAs that were downregluated with LBRD standard decoction treatment, and red symbols denote up-regulated mRNAs. Their negative regulation of mRNAs by miRNAs is represented by a grey line. B The specific binding regions between miR-144-3p and Gad1/VGAT, and miR-495 and Bdnf were obtained from the online software database. C, D The relationship between miR-144-3p and Gad1/VGAT, and miR-495 and Bdnf were confirmed by dual-luciferase reporter and RIP assays (n = 3). Normal immunoglobulin was used as a negative control. E, F qRT-PCR analysis of Gad1, VGAT, and Bdnf in PC12 cells respectively transfected with miR-144-3p and miR-495 mimics or inhibitor. G Association of miR-144-3p, miR-495 and its corresponding target genes were assessed by Pearson’s correlation coefficients with qRT-PCR results (n = 9 pre/group). Blue dots indicate congregation of the LBRD standard decoction-treated group and red dots indicate congregation of the depression group. H The expression level of Gad1, VGAT, and BDNF proteins were measured in mPFCs tissues obtained from LBRD standard decoction-treated group. ^**P < 0.01, ^***P < 0.001, compared to control group; ^##P < 0.01, ^###P < 0.001, compared to CUMS + Saline group. CUMS + Saline: CUMS + Saline group, CUMS + LBRD: CUMS + Lily Bulb and Rehmannia Decoction group