Anti-inflammatory properties of the ethanol extract from Clerodendrum cyrtophyllum Turcz based on in vitro and in vivo studies

Abstract

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.

Developmental toxicity of Clerodendrum cyrtophyllum turcz ethanol extract in zebrafish embryo

“…After the extraction, the EE was analysed with various analytical techniques including colorometric methods, HPLC-DAD-Orbitrap-MS analysis. The concentration of total phenolic compounds and flavonoid in the extract were 23.3 ± 1.5 GAE mg/g and 2.97 ± 0.01 QE mg/g expressed in dry weight of leaves material (Nguyen et al, 2020b).…”

“…Preparation of total extract: The ethanol extract from leaves of Clerodendrum cyrtophyllum Turcz was prepared as described in our previous study (Nguyen et al, 2020b). After the extraction, the EE was analysed with various analytical techniques including colorometric methods, HPLC-DAD-Orbitrap-MS analysis.…”

“…Clerodendrum cyrtophyllum Turcz (Lamiaceae), a medicinal plant, is widely used in traditional medicine of many Asian countries such as China, India, Japan, Korea, Thailand, and Vietnam for the treatment of colds, high fever, migraines, hypertension, enteritis, dyspepsia, inflammation of the throat, rheumatic arthritis, fever, jaundice, leukorrhea, syphilis, and typhoid (Kar et al, 2014;Zhou et al, 2013). Several therapeutic and pharmacological properties of Clerodendrum cyrtophyllum Turcz have recently been reported such as antioxidant, anti-inflammatory (Nguyen et al, 2020a(Nguyen et al, , 2020b, and anticancer (Cheng et al, 2001) activities. Despite their frequent use, studies on its toxicity and its effects on reproductive tissues or embryos are currently unknown.…”

Evaluation of Aster bakerianus Burtt Davy ex C.A. Sm. Crude Root Extract for Acute Antiinflammatory Activity in Rats

“…Acute inflammation is a rapid, short-lived initial response of tissue to harmful stimuli (such as injury) initiated by cells constituting the relevant tissues (Ambriz-Pérez et al, 2016) and is characterised by accumulation of fluid, plasma proteins and the leukocytes which release a large number of soluble inflammatory mediators which are responsible for the initiation, progression, persistence, modulation (regulation) and eventual resolution of the acute state of inflammation (Oguntibeju, 2018;Nguyen et al, 2020). The classical signs of acute inflammation include oedema, erythema, pain, heat, and primarily loss of function of the affected part of the body (Nathan, 2002;Husein et al, 2012).…”

“…Numerous proinflammatory mediators are released during an inflammatory response, including the vasoactive amines; histamine, serotonin, bradykinin (Kulinsky, 2007;El-Shitany et al, 2014) and the cytokines, interleukin 1 β (IL-1 β), IL-6, IL-8, IL-12, tumour necrosis factor-α (TNF-α) and interferon-γ (INF-γ) as well as prostaglandins especially PGE2 through cyclooxygenase-2 (COX-2), leukotrienes through lipooxygenase (LOX), nitric oxide (NO) through the inducible nitric oxide synthase (iNOS) (Husein et al, 2012; Kulinsky, 2007;El-Shitany et al, 2014). Also released is the nuclear factor kappa B (NF-κB), a transcription factor that plays an important role in the transcription of genes, as it induces transcription of its target genes such as COX-2, iNOS, TNF-α, IL-1β, and IL-6, chemokines and adhesion molecules (Karin & Ben-Neriah, 2000) that cause acute inflammation (Nguyen et al, 2020). The cytokines play major roles in the initiation and amplification of inflammatory processes (Calixto et al, 2004).…”