In Vitro Immunomodulatory Effects of Inonotus obliquus Extracts on Resting M0 Macrophages and LPS-Induced M1 Macrophages
Dayue Shen, Yating Feng, Xilan Zhang, Jing Liu, Le Gong, Hui Liao, Rongshan Li
Abstract
Background. Inonotus obliquus (Chaga) is a parasitic fungus that is distributed mainly in northeast China. Our literature research showed chaga polysaccharides have bilateral effects on tumor necrosis factor (TNF)-α and interleukin (IL)-1β levels when they exert antitumor and antidiabetic activities. The current research tried to explore the influence of chaga extracts on inflammatory factors via macrophage polarization which has bilateral immune-regulation not only on healthy tissue homeostasis but also on pathologies. Methods. Chaga was extracted with 100°C water and precipitated with 80% ethanol. The extracts were studied on RAW264.7 macrophage at resting condition (M0) and lipopolysaccharide (LPS)-activated subtype (classic activated macrophage, M1). The IL-1β, TNF-α, nitric oxide (NO) level, and the protein expressions of M1 and alternative activated macrophage (M2) markers including IL-1β, inducible NO synthase (iNOS), mannose receptor (CD206), and arginase (Arg)-1 were compared. Results. The 100 g extracts contained 13.7 g polysaccharides and 1.9 g polyphenols. Compared with M0, the 50 μg/mL extracts increased NO level ( <a:math xmlns:a="http://www.w3.org/1998/Math/MathML" id="M1"> <a:mi>P</a:mi> <a:mo><</a:mo> <a:mn>0.05</a:mn> </a:math> ) and decreased CD206 and Arg-1 expression significantly ( <c:math xmlns:c="http://www.w3.org/1998/Math/MathML" id="M2"> <c:mi>P</c:mi> <c:mo><</c:mo> <c:mn>0.05</c:mn> </c:math> ). The extracts at 100–200 μg/mL increased NO and TNF-α level ( <e:math xmlns:e="http://www.w3.org/1998/Math/MathML" id="M3"> <e:mi>P</e:mi> <e:mo><</e:mo> <e:mn>0.05</e:mn> </e:math> ), but increased iNOS and IL-1β expression significantly ( <g:math xmlns:g="http://www.w3.org/1998/Math/MathML" id="M4"> <g:mi>P</g:mi> <g:mo><</g:mo> <g:mn>0.05</g:mn> </g:math> ). Compared with M1, the extracts decreased NO level at 25, 50, 100, and 200 μg/mL and decreased IL-1β and TNF-α level at 100–200 μg/mL significantly ( <i:math xmlns:i="http://www.w3.org/1998/Math/MathML" id="M5"> <i:mi>P</i:mi> <i:mo><</i:mo> <i:mn>0.05</i:mn> </i:math> ). At 25–200 μg/mL, the extracts significantly increased CD206 and Arg-1 expression and decreased IL-1β and iNOS expression separately ( <k:math xmlns:k="http://www.w3.org/1998/Math/MathML" id="M6"> <k:mi>P</k:mi> <k:mo><</k:mo> <k:mn>0.05</k:mn> </k:math> ). Conclusions. Our research suggested that the bilateral effects of the chaga extracts on iNOS, IL-1β, and NO level on M0/M1 macrophages might be related with chaga polysaccharides and chaga polyphenols. Some in vivo anticancer and antidiabetic research of purified chaga polysaccharides related to macrophage differentiation should be conducted further.