Abstract
The extract of Citrus reticulata has been studied for its biological activities, due to its citrus flavonoid content. The extract and its flavonoid compounds exhibit growth inhibition property in several cancer cell lines and in vivo models. Conversely, the extract can also induce cell proliferation and angiogenesis, and shows estrogenic effects by in vitro and in vivo. Because of the contrasting effects that depend on the concentration or dosage, the precise action of the extract and its flavonoids need to be elucidated in various cell types. The objective of this study is to evaluate the effect of Citrus reticulata peel extract (Citrus extract) and hesperidin, a citrus flavonoid, on the modulation of cell proliferation in the RAW 264.7 macrophage cell line. Cell viability under Citrus extract or hesperidin treatment was assessed by using the MTT assay. The expression of interleukin-10 (IL-10), an anti-inflammatory cytokine, modulated by Citrus extract was also examined by immunostaining. Low concentrations of Citrus extract at 1 and 100μg/mL were able to induce cell proliferation, although in significant, as shown by cell viability of 138 and 114%, respectively. At higher concentrations of 500, 750, and 1000μg/mL, Citrus extract decreased cell viability significantly by up to 64, 46, and 36%, respectively. Accordingly, hesperidin at low (3.1μg/mL −61.1μg/mL) concentration increased cell viability significantly by up to 116-136% where as high (152.6203μg/mL–305.3203μg/mL) concentration reduced cell viability significantly by up to 10-61%. The value of the 50% inhibitory concentration (IC50) of Citrus extract was more than three times higher (756μg/mL) than that of hesperidin (203μg/mL = 332μM). Additionally, 250μg/mL of Citrus extract was able to induce IL-10 expression compared to control. These results demonstrated that Citrus extract and hesperidin exerted a biphasic effect on macrophage cells. The future development of Citrus extract as a co-chemotherapeutic, anticancer, or immunomodulatory agent should include careful consideration of its biphasic effect on each cell type
Highlights
Citrus plants have attracted researchers’ attention due to their biological activities since the early 18th century (Manthey et al, 2001)
We evaluate the effect of ethanolic extract from Citrus reticulata peel (Citrus extract) and hesperidin at various concentrations on the modulation of cell proliferation in the RAW 264.7 macrophage cell line
We reveal that both Citrus extract and hesperidin show a biphasic effect on RAW 264.7 macrophage cell viability
Summary
Citrus plants (family: Rutaceae) have attracted researchers’ attention due to their biological activities since the early 18th century (Manthey et al, 2001). Their phytochemical healthpromoting properties are mainly based on the antioxidant activities of flavonoid compounds that contribute to the cardiovascular disease and cancer prevention, anti-inflammatory, antiviral, and antimicrobial properties of citrus (Barreca et al, 2017; Benavente-Garcia et al, 1997). In vitro assay in MCF-7 Increases selectivity for (Yunas et al, extract breast cancer cell line doxorubicin 2007). In vitro assay in WiDr Induces cell proliferation and 10-500μg/mL
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