Enhanced rosmarinic acid biosynthesis in <i>Solenostemon scutellarioides</i> culture: a precursor-feeding strategy

<div><p>The aim of this study was to employ precursor-feeding strategy for the improved production of rosmarinic acid (RA) in <i>S</i><i>olenostemon</i><i> scutellarioides in vitro</i>. The cultures were fed with precursors, namely l-phenylalanine (Phe), l-tyrosine (Tyr) and cucumber juice (CJ), at different concentrations. Phe (100 mg L^− 1) and Tyr (400 mg L^− 1) caused ∼1.5- and 2.1-fold increase in RA accumulation within 48 h. CJ (50 mg L^− 1) feeding displayed highest RA content (∼1.6-fold) in 72 h. In this study, we focused on the function of individual precursor on key enzymes involved in RA biosynthesis. The phenylalanine ammonia lyase activity was significantly upregulated after Phe (100 mg L^− 1) feeding, while tyrosine aminotransferase and hydroxyphenylpyruvate reductase activities were improved with Tyr (400 mg L^− 1) treatment. However, rosmarinic acid synthase activity was significantly enhanced by all three precursors. In synergy study, Phe (100 mg L^− 1) + Tyr (400 mg L^− 1) could enhance (∼3.1-fold) RA biosynthesis within 48 h.</p></div

Additional file 1: Figure S1. of Pharmacological basis of the use of the root bark of Zizyphus nummularia Aubrev. (Rhamnaceae) as anti-inflammatory agent

The effect of octadecahydro-picene-2,3,14,15-tetranone on the viability of RAW 264.7 cells. (PPTX 47 kb

A Schematic overview of <i>in vivo</i> experimental protocol.

<p>A Schematic overview of <i>in vivo</i> experimental protocol.</p

A schematic overview of the hypothesis developed in this study regarding probable protective mechanism of SR against diabetic cardiomyopathy.

<p>Green dotted lines represented the restricted pathological events by SR.</p

Effect of SR on blood glucose^a, serum insulin, HOMA-IR and HOMA-β.

<p>Data were expressed as mean ± SD (n = 6). ^#p < 0.01 compared with Group I; *p < 0.05 compared with Group II; **p < 0.01 compared with Group II. Group I: Normal control; Group II: T2D control, Group III: T2D rats treated with SR (50 mg/kg, orally); Group IV: T2D rats treated with SR (100 mg/kg, orally); Group V: T2D rats treated with glibenclamide (1 mg/kg, orally). ¹HOMA-IR = [(Fasting serum insulin in U/l x Fasting blood glucose in mmol/l)/22.5] ²HOMA-β = (Fasting serum insulin in U/l x 20/Fasting blood glucose in mmol/l– 3.5) ^a The blood glucose levels used in these assessments were estimated 24 h before sacrificing the animals. Considering the overall duration of the experiment, it has been postulated that the glucose concentration will not vary significantly within 24 h after 28 days of post-treatment.</p

Effect of SR on ATP level, NAD level, DNA fragmentation and DNA oxidation in the myocardial tissues of T2D rats.

<p>Data were expressed as mean ± SD (n = 6). ^#p < 0.01 compared with Group I; *p < 0.05 compared with Group II; **p < 0.01 compared with Group II. Group I: Normal control; Group II: T2D control, Group III: T2D rats treated with SR (50 mg/kg, orally); Group IV: T2D rats treated with SR (100 mg/kg, orally); Group V: T2D rats treated with glibenclamide (1 mg/kg, orally).</p

Effect on protein carbonylation, ROS production, lipid peroxidation and co-enzyme Q9 levels in the absence (Pb-acetate) and existence of AEEF (Pb-acetate +AEEF) in experimental mice.

<p>Results were represented as mean ± SE (n = 6). ^$ Results differed significantly (p < 0.05) from Pb-acetate control. ^#Results differed (p < 0.01) significantly from normal control. * Results significantly (p < 0.05) differed from Pb-acetate control. ** Results differed (p < 0.01) significantly from Pb-acetate control.</p

Histological assessments of testes along with histo-quantification data of experimental mice in the absence (Pb-acetate) and existence of AEEF (Pb-acetate + AEEF).

<p>Histological sections 100 x (Panel A) and 400 x (Panel B) of testes. The section of testes of normal control mice exhibited all stages of spermatogenesis, while, testes of Pb-acetate treated mice showed disruption of normal arrangement of seminiferous tubules also in the process of spermagenesis. Yellow arrows represented spermatogonia close to the basement membrane; red arrow represented primary spermatocytes; green arrows represented round spermatids; blue arrows denote elongated spermatids; black arrows represented complete spermatozoa. However, AEEF treatment could attenuate the Pb-acetate mediated toxic manifestations in testes of mice. Panel C. The Johnsen score was measured (400 X, comprising one seminiferous tubule). Seminiferous tubule at Johnsen score 10 presenting all stages of spermatogenesis. The Jhonsen score is descending with the toxic occurrence within testicular tissues. Results were expressed as mean ± SE, (n = 60). ^# Results differed significantly (p < 0.01) from normal control. ^**Results significantly (p < 0.01) differed from Pb-acetate control.</p

Effect on Pb accumulation (Panel A), DNA fragmentation (Panel B) DNA oxidation (Panel C) and ATP levels (Panel D) in the absence (Pb-acetate) and existence of AEEF (Pb-acetate +AEEF) in heart, kidney, liver, brain and testes in mice.

<p>Results were denoted as mean ± SE (n = 6). ^#Results differed significantly (p < 0.01) from normal control. * Results significantly (p < 0.05) differed from Pb-acetate control. ** Results significantly (p < 0.01) differed from Pb-acetate control.</p

Schematic presentation of the hypothesis developed in this study regarding the overall protective mechanism of AEEF against Pb toxicity.

<p>The dark blue arrows indicate the cellular events involved in Pb-induced pathophysiologies. The green lines denoted the activity promoted (+) by AEEF, while, red lines denoted the activity restricted (-) by AEEF.</p