Anti–Inflammatory and Antioxidant Properties of the Ethanol Extract of Clerodendrum Cyrtophyllum Turcz in Copper Sulfate-Induced Inflammation in Zebrafish

Abstract

Oxidative stress and inflammation are commonly present in many chronic diseases. These responses are closely related to pathophysiological processes. The inflammatory process can induce oxidative stress and vice versa through the activation of multiple pathways. Therefore, agents with antioxidant and/or anti-inflammatory activities are very useful in the treatment of many pathologies. Clerodendrum cyrthophyllum Turcz, a plant belonging to the Verbenaceae family, is used in Vietnamese traditional medicine for treating migraine, hypertension, inflammation of the throat, and rheumatic arthritis. Despite its usefulness, studies on its biological properties are still scarce. In this study, ethanol extract (EE) of leaves of C. cyrtophyllum showed protective activity against CuSO4 toxicity. The protective activity was proven to relate to antioxidant and anti-inflammatory properties. EE exhibited relatively high antioxidant activity (IC50 of 16.45 µg/mL) as measured by DPPH assay. In an in vivo anti-antioxidant test, three days post fertilization (dpf) zebrafish larvae were treated with different concentrations of EE for 1 h and then exposed to 10 µM CuSO4 for 20 min to induce oxidative stress. Fluorescent probes were used to detect and quantify oxidative stress by measuring the fluorescent intensity (FI) in larvae. FI significantly decreased in the presence of EE at 5 and 20 µg/mL, demonstrating EE’s profound antioxidant effects, reducing or preventing oxidative stress from CuSO4. Moreover, the co-administration of EE also protected zebrafish larvae against oxidative damage from CuSO4 through down-regulation of hsp70 and gadd45bb expression and upregulation of sod. Due to copper accumulation in zebrafish tissues, the damage and oxidative stress were exacerbated overtime, resulting in the upregulation of genes related to inflammatory processes such as cox-2, pla2, c3a, mpo, and pro- and anti-inflammatory cytokines (il-1ß, il-8, tnf-α, and il-10, respectively). However, the association of CuSO4 with EE significantly decreased the expression of cox-2, pla2, c3a, mpo, il-8, and il-1ß. Taken together, the results suggest that EE has potent antioxidant and anti-inflammatory activities and may be useful in the treatment of various inflammatory diseases.

Molecular Mechanisms Underlying Protective Role of Quercetin on Copper Sulfate-Induced Nephrotoxicity in Mice

“…Inflammation plays a critical role in the progression of CuSO 4 exposure-induced toxic effects ( 54 – 56 ). In the present study, our results showed that CuSO 4 exposure significantly upregulated the expression of IL-1β, TNF-α, and IL-6 proteins and mRNAs in the kidney tissues.…”

Anti-Inflammatory Activity of Exopolysaccharides from Phormidium sp. ETS05, the Most Abundant Cyanobacterium of the Therapeutic Euganean Thermal Muds, Using the Zebrafish Model

“…Exposure to copper can cause injuries to superficial tissues, in particular to the hair cells of the zebrafish lateral line, followed by an acute inflammatory response [49,50]. Thus, treatment of zebrafish embryos/larvae with copper is today commonly used to analyze inflammatory response [65,66]. Since cyanobacterial EPS can bind dissolved heavy metal bipositive cations such as Cu 2+ or Pb 2+ [51], Phormidium EPS was applied to copper-treated larvae only after multiple washes.…”

Developmental toxicity of Clerodendrum cyrtophyllum turcz ethanol extract in zebrafish embryo

“…Quantitative PCR Total RNA extraction, DNase and reverse transcription Total RNA extraction, DNase and reverse transcription were carried out as described in previous our study (Nguyen et al, 2020a) using Trizol Reagent solution (Ambion, Thermofisher Scientific), DNA-free™ DNA Removal Kit-Invitrogen and the RevertAid RT kit (Thermo Scientific).…”

“…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.…”