The effects of TDBP-TAZTO (5 and 50 mg/kg) on apoptosis-related proteins in the hippocampus.

<p>The results are presented as mean ± SE (n = 5/group). *<i>p</i><0.05, vs. control group. The O.D. values were displayed in Bax (B), Bcl-2 (C) and cleaved caspase-3 (D) normalized to β-actin.</p

DataSheet_1_Transcriptome analysis reveals that jasmonic acid biosynthesis and signaling is associated with the biosynthesis of asperosaponin VI in Dipsacus asperoides.docx

Dipsacus asperoides is a perennial herb, the roots of which are abundant in asperosaponin VI, which has important medicinal value. However, the molecular mechanism underlying the biosynthesis of asperosaponin VI in D. asperoides remains unclear. In present study, a comprehensive investigation of asperosaponin VI biosynthesis was conducted at the levels of metabolite and transcript during root development. The content of asperosaponin VI was significantly accumulated in two-leaf stage roots, and the spatial distribution of asperosaponin VI was localized in the xylem. The concentration of asperosaponin VI gradually increased in the root with the development process. Transcriptome analysis revealed 3916 unique differentially expressed genes (DEGs) including 146 transcription factors (TFs) during root development in D. asperoides. In addition, α-linolenic acid metabolism, jasmonic acid (JA) biosynthesis, JA signal transduction, sesquiterpenoid and triterpenoid biosynthesis, and terpenoid backbone biosynthesis were prominently enriched. Furthermore, the concentration of JA gradually increased, and genes involved in α-linolenic acid metabolism, JA biosynthesis, and triterpenoid biosynthesis were up-regulated during root development. Moreover, the concentration of asperosaponin VI was increased following methyl jasmonate (MeJA) treatment by activating the expression of genes in the triterpenoid biosynthesis pathway, including acetyl-CoA acetyltransferase (DaAACT), 3-hydroxy-3-methylglutaryl coenzyme A synthase (DaHMGCS), 3-hydroxy-3-methylglutaryl coenzyme-A reductase (DaHMGCR). We speculate that JA biosynthesis and signaling regulates the expression of triterpenoid biosynthetic genes and facilitate the biosynthesis of asperosaponin VI. The results suggest a regulatory network wherein triterpenoids, JA, and TFs co-modulate the biosynthesis of asperosaponin VI in D. asperoides.</p

The effects of TDBP-TAZTO (5 and 50 mg/kg) on spatial learning and memory in rats in the Morris water test.

<p>(A) In the acquisition trial, TDBP-TAZTO rats showed an increased latency to the platform. (B) In the probe trial, TDBP-TAZTO rats had a decreased time spent in the target quadrant. (C) From day 1 to day 6, TDBP-TAZTO did not affect the swimming speed compared with that in control group. Data ate expressed as mean ± SE (n = 10/group). ^a<i>p</i> < 0.05, TDBP-TAZTO 5 mg/kg group vs. control group; ^b<i>p</i> < 0.05, TDBP-TAZTO 50 mg/kg group vs. control group.</p

Table_1_Transcriptome analysis reveals that jasmonic acid biosynthesis and signaling is associated with the biosynthesis of asperosaponin VI in Dipsacus asperoides.xlsx

Dipsacus asperoides is a perennial herb, the roots of which are abundant in asperosaponin VI, which has important medicinal value. However, the molecular mechanism underlying the biosynthesis of asperosaponin VI in D. asperoides remains unclear. In present study, a comprehensive investigation of asperosaponin VI biosynthesis was conducted at the levels of metabolite and transcript during root development. The content of asperosaponin VI was significantly accumulated in two-leaf stage roots, and the spatial distribution of asperosaponin VI was localized in the xylem. The concentration of asperosaponin VI gradually increased in the root with the development process. Transcriptome analysis revealed 3916 unique differentially expressed genes (DEGs) including 146 transcription factors (TFs) during root development in D. asperoides. In addition, α-linolenic acid metabolism, jasmonic acid (JA) biosynthesis, JA signal transduction, sesquiterpenoid and triterpenoid biosynthesis, and terpenoid backbone biosynthesis were prominently enriched. Furthermore, the concentration of JA gradually increased, and genes involved in α-linolenic acid metabolism, JA biosynthesis, and triterpenoid biosynthesis were up-regulated during root development. Moreover, the concentration of asperosaponin VI was increased following methyl jasmonate (MeJA) treatment by activating the expression of genes in the triterpenoid biosynthesis pathway, including acetyl-CoA acetyltransferase (DaAACT), 3-hydroxy-3-methylglutaryl coenzyme A synthase (DaHMGCS), 3-hydroxy-3-methylglutaryl coenzyme-A reductase (DaHMGCR). We speculate that JA biosynthesis and signaling regulates the expression of triterpenoid biosynthetic genes and facilitate the biosynthesis of asperosaponin VI. The results suggest a regulatory network wherein triterpenoids, JA, and TFs co-modulate the biosynthesis of asperosaponin VI in D. asperoides.</p

The effects of TDBP-TAZTO (5 and 50 mg/kg) on the locomotor activity (A) and the durations of immobility in FST (B) in rats.

<p>The results are presented as mean ± SE (n = 10/group). *<i>p</i> < 0.05, vs. control group.</p

Representative immunohistochemical location of GFAP in DG (A-C), CA3 (D-F) and CA1 regions (G-I) of the hippocampus (200×).

<p>The sections from control group (A, D and G), TDBP-TAZTO 5 mg/kg group (B, E and H) and 50 mg/kg group (C, F and I). The arrows indicates astrocytes. DG, CA3 and CA1 areas had some small dispersed non-branched astrocytes in the control rats (A, D and G, respectively), while small or large branched astrocytes in 5 mg/kg (B, E and H, respectively) and 50 mg/kg rats (C, F and I, respectively).</p

Representative HE staining of adrenal gland (200 ×).

<p>Sections from control group (A), TDBP-TAZTO 5 mg/kg group (B) and TDBP-TAZTO 50 mg/kg group (C). The arrows indicates zona fasciculata. The regularly arranged long columnar cells without cell hyperplasia or hypertrophy were observed in the zona fasciculata in the control group (A). Main diffuse adrenal hyperplasia and hypertrophy of the zona fasciculate were noted in 5 and 50 mg/kg groups (B and C).</p

The effects of TDBP-TAZTO (5 and 50 mg/kg) on BDNF and SYP expression in hippocampus.

<p>The results are presented as mean ± SE (n = 5/group). *<i>p</i><0.05, vs. control group. BDNF (A) was expressed as pg/mg protein. The O.D. values were displayed in SYP (B) normalized to β-actin.</p

The effects of TDBP-TAZTO (5 and 50 mg/kg) on plasma ACTH (A) and serum CORT (B).

<p>The results are presented as mean ± SE (n = 10/group). *<i>p</i> < 0.05, vs. control group.</p

Morris Water Maze test.

<p>Note: south (S), west (W), east (E), northwest (N), northeast (NE), southeast (SE), northwest (NW) and southwest (SW).</p><p>Morris Water Maze test.</p