Ethnopharmacological relevance: Clerodendrum cyrtophyllum Turcz, a plant belonging to the Verbenaceae family, has been used in traditional medicine for the treatment of various inflammatory diseases in many Asian countries.Aim of the study: The study aimed to evaluate anti-inflammatory properties of the ethanol extract from Clerodendrum cyrthophyllum Turcz leaves (EE-CC) through in vitro and in vivo models. Material and methods: Total phenolic and flavonoid contents in the extract were determined using colorimetric methods and HPTLC. In red blood cell membrane stabilization model, rat erythrocyte suspension was treated with crude ethanol extract at different concentrations, the hemoglobin content of the supernatant solution released by red blood hemolysis was estimated. We also evaluated the effects of the ethanol extract from this plant on the production of nitric oxide (NO), tumor necrosis factor alpha (TNF-α) in stimulated RAW 264.7 cells. In order to elucidate its anti-inflammatory molecular mechanisms, we further evaluated the effects of the EE-CC on the expression of the inflammatory genes in inflammation-induced zebrafish model by tail-cutting using qPCR analysis. Results: Colorimetric methods and HPTLC revealed high phenolic and flavonoid contents in the extract. In the red blood cell membrane stabilization model, the amount of hemoglobin released by red blood hemolysis significantly decreased in the presence of EE-CC, demonstrating a strong membrane stabilizing activity. EE-CC did not cause any toxic effect on cell viability but strongly inhibited NO and TNF-ɑ release due to LPS induction. The association with EE-CC significantly reduced the expression of cox-2, pla2, c3a, il-1(il1fma), il-8 (cxcl8b.1), tnf-α, and nf-ƙb, while increased the expression of the anti-inflammatory cytokine il-10 gene in cut-tail induced inflammation of zebrafish model. Conclusions: Taken together, the results suggest that the raw ethanol extract from C. cyrtophyllum Turcz leaves presents potent anti-inflammatory activities and may be useful for the treatment of various inflammatory diseases.

Clerodendrum cyrtophyllum Turcz has been used in traditional medicine for the treatment of various diseases. In spite of its therapeutic applications, research on its toxicity and teratogenicity is still limited. Aim of the study: The study aimed to investigate the developmental toxicity of the ethanol extract of C. cyrtophyllum (EE) in zebrafish embryo model. Material and methods: Major compounds from crude ethanol extract of Clerodendron cyrtophyllum Turcz leaves were determined using HPLC-DAD-Orbitrap-MS analysis. The developmental toxicity of EE were investigated using zebrafish embryo model. Zebrafish embryos at 6 h post-fertilization (hpf) were treated with EE at different concentrations. Egg coagulation, mortality, hatching, yolk sac edema, pericardial edema and teratogenicity were recorded each day for during a 5-day exposure. At time point 120 hpf, body length, pericardial area, heartbeat and yolk sac area were assessed. In order to elucidate molecular mechanisms for the developmental toxicity of EE, we further evaluated the effects of the EE on the expression of genes involved on signaling pathways affecting fish embryo's development such as heart development (gata5, myl7, myh6, has2, hand2, nkx 2.5), oxidative stress (cat, sod1, gpx4, gstp2), wnt pathway (β-catenin, wnt3a, wnt5, wnt8a, wnt11), or cell apoptosis (p53, bax, bcl2, casp3, casp8, casp9, apaf-1, gadd45bb) using qRT-PCR analysis. Results: Our results demonstrated that three major components including acteoside, cirsilineol and cirsilineol-4'-O-β-D-glucopyranoside were identified from EE. EE exposure during 6-96 h post-fertilization (hpf) at doses ranging from 80 to 200 μg/mL increased embryo mortality and reduced hatching rate. EE exposure at 20 and 40 μg/mL until 72-120 hpf induced a series of malformations, including yolk sac edema, pericardial edema, spine deformation, shorter body length. Based on two prediction models using a teratogenic index (TI), a 25% lethality concentration (LD25) and the no observed-adverse-effect level (NOAEL), EE is considered as teratogenic for zebrafish embryos with TI (LC50/EC50) and LD25/NOAEC values at 96 hpf reaching 3.87 and 15.73 respectively. The mRNA expression levels of p53, casp8, bax/bcl2, gstp2, nkx2.5, wnt3a, wnt11, gadd45bb and gata5 were significantly upregulated by EE exposure at 20 and 40 μg/mL while the expression of wnt5, hand2 and bcl2 were downregulated.

Danshen (Salvia miltiorrhiza Bunge) is one of the most used medicinal plants in the Oriental medicine and has been well studied for application in modern medicine. In our continuing study on chemical constituents of danshen cultivated in Vietnam, using chromatography separation resulted in the isolation of six phenolic compounds including a benzophenone, iriflophenone 2-O-α-L-rhamnopyranoside (1), and five phenolic acids including rosmarinic acid (2), rosmarinic acid methyl ester (3), rosmarinic acid ethyl ester (4), salvianolic acid A methyl ester (5) and salvianolic acid A ethyl ester (6) from the butanol portion of the danshen crude extract. Beside the typically main phenolic acid components, to our knowledge, iriflophenone 2-O-α-L-rhamnopyranoside (1) was first isolated from salvia sp. On biological testing, compound 1 showed strong antiproliferative activity on HL-60 leukemia cells with the IC 50 of 8.9 µM; compounds 1 and 3-6 inhibited markedly nitric oxide production in lipopolysaccharide-treated RAW 264.7 cells.

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The study was conducted to determine the effects of different levels of dietary SID methionine on growth performance, carcass yield, meat quality, myogenic gene expression of two coloured chicken groups. A total of 480 one-day-old chicks (240 Ri hybrid and 240 Luong Phuong) of uniform body weight was divided into a 2 × 3 factorial arrangement of treatments with four replicate cages of 20 chicks per treatment. Two factors include 2 chicken groups (Ri hybrid-slow-growing strain and Luong Phuong-fast-growing strain) and three dietary SID methionine concentration (low, adequate, high). The methionine levels were set up by the recommendation of Evonik (2010) for coloured chickens. At 12 weeks of age, 8 male and 8 female chicks per treatment were randomly chosen for the carcass yield, meat quality, and myogenic gene expression evaluation (myostatin, myf5, MEF2B). The results showed that the higher level of dietary SID methionine than that of Evonik recommendation (2010) improved growth performance in slow-growing colored chickens but had no effect on body weight gain in fast-growing strain. The increasing breast muscle yield and decreasing abdominal fat were observed in the fast-growing chicks fed high dietary SID methionine. The improvements in breast muscle of the fast-growing colored strain was reflected by increased mRNA expression of Myf5 and MEF2B but decreased expression of myostatin.