To identify the immune activity of different molecular weight components of Colla corii asini, it was separated into three components: <10 (F1), 10-30 (F2), and >30 kDa (F3) by Superdex 75pg dextran gel column, and their abilities to stimulate cells to release NO and TNF-α were compared. Then the fraction with the strongest immunomodulatory ability was used to study the activation effect on macrophages and its mechanism. Results showed that F2 could stimulate macrophages to release more NO and TNF-α than F1 and F3 at the same concentration. In addition, it could activate RAW264.7 cells and promote the release of ROS and IL-6. The results of q-PCR showed that F2 could promote the expression of TNF-α, IL-6, iNOS and TLR4 genes in cells and bind to TLR4 receptors. The phosphorylation of JNK, ERK and P38 proteins was promoted, which indicated that F2 played an immunomodulatory role through the MAPK signaling pathway. This research will provide theoretical guidance for optimizing the production and processing conditions of Colla corii asini to produce high-quality product.
ZHANG Guowei
,
MA Junhua
,
LIANG Yujing
,
WANG Zhaojun
,
ZENG Maomao
,
CHEN Jie
,
QIN Fang
,
HE Zhiyong
. Immunomodulatory effects of different molecular weight fractions of Colla corii asini on RAW264.7 mouse macrophages[J]. Food and Fermentation Industries, 2022
, 48(14)
: 125
-131
.
DOI: 10.13995/j.cnki.11-1802/ts.029626
[1] HACHIMURA S, TOTSUKA M, HOSONO A.Immunomodulation by food:Impact on gut immunity and immune cell function[J].Bioscience, Biotechnology, and Biochemistry, 2018, 82(4):584-599.
[2] BATATINHA H A P, BIONDO L A, LIRA F S, et al.Nutrients, immune system, and exercise:Where will it take us?[J].Nutrition, 2019,61:151-156.
[3] 刘良初. 阿胶成分对比分析研究[J].中成药研究, 1983(1):36-37.
LIU L C.Comparative analysis of ejiao components[J].Chinese Patent Medicine Research, 1983(1):36-37.
[4] 张珣, 王静凤, 李冰, 等.阿胶对小鼠免疫功能的影响[J].食品工业科技, 2011, 32(11):400-402;433.
ZHANG X, WANG J F, LI B, et al.Effect of Colla corii asini on immune function in mice[J].Science and Technology of Food Industry, 2011, 32(11):400-402;433.
[5] 韩晶. 阿胶通过抵抗免疫细胞凋亡起到免疫保护的作用[D].北京:北京协和医学院, 2018.
HAN J.The traditional Chinese medicine ejiao is an immunoprotective agent against apoptosis[D].Beijing:Peking Union Medical College, 2018.
[6] 樊绘曾, 刘曦, 徐天泽.驴皮成分在化胶过程中的降解[J].中国中药杂志, 1994(9):543-545.
FAN K Z, LIU X, XU T Z.Degradation of the constituents in solubilization process of donkey skin[J].Chinese Journal of Traditional Chinese Medicine, 1994(9):543-545.
[7] GORNALL A G, BARDAWILL C J, DAVID M M.Determination of serum proteins by means of the biuret reaction[J].Journal of Biological Chemistry, 1949, 177(2):751-766.
[8] ZHANG H, YAO H Y, CHEN F.Purification and characterization of glutamate decarboxylase from rice germ[J]. Food Chemistry, 2017,101(4):1 670-1 676.
[9] YANG Q, CAI X X, HUANG M C, et al.A specific peptide with immunomodulatory activity from Pseudostellaria heterophylla and the action mechanism[J].Journal of Functional Foods, 2020, 68:103887.
[10] LIN Z H, HU J S, SHI H G, et al.Water extract of medicinal ink (WEMI) attenuates lipopolysaccharide-induced NO production of RAW264.7 cells via downregulating JAK2/STAT3-mediated iNOS expression[J].Journal of Ethnopharmacology, 2022, 282:114636.
[11] GEUM N G, EO H J, KIM H J, et al.Immune-enhancing activity of Hydrangea macrophylla subsp.serrata leaves through TLR4/ROS-dependent activation of JNK and NF-κB in RAW264.7 cells and immunosuppressed mice[J].Journal of Functional Foods, 2020, 73:104139.
[12] YANG Q, HUANG M C, CAI X X, et al.Investigation on activation in RAW264.7 macrophage cells and protection in cyclophosphamide-treated mice of Pseudostellaria heterophylla protein hydrolysate[J].Food and Chemical Toxicology, 2019, 134:110816.
[13] AGGARWAL B B, GUPTA S C, KIM J H.Historical perspectives on tumor necrosis factor and its superfamily:25 years later, a golden journey[J].Blood, 2012, 119(3):651-665.
[14] CHALAMAIAH M, HEMALATHA R, JYOTHIRMAYI T, et al.Chemical composition and immunomodulatory effects of enzymatic protein hydrolysates from common carp (Cyprinus carpio) egg[J].Nutrition, 2015, 31(2):388-398.
[15] GARBERS C, HEINK S, KORN T, et al.Interleukin-6:Designing specific therapeutics for a complex cytokine[J].Nature Reviews Drug Discovery, 2018, 17(6):395-412.
[16] 林诺怡, 成坚, 王琴, 等.柚皮蛋白的结构表征及细胞免疫活性初步研究[J].食品与发酵工业, 2021, 47(3):59-65.
LIN N Y, CHENG J, WANG Q, et al.A preliminary study on structure and cellular immune activity of pomelo peel protein[J].Food and Fermentation Industries, 2021, 47(3):59-65.
[17] TALAPPHET N, PALANISAMY S, LI C S, et al.Polysaccharide extracted from Taraxacum platycarpum root exerts immunomodulatory activity via MAPK and NF-κB pathways in RAW264.7 cells[J].Journal of Ethnopharmacology, 2021, 281:114519.
[18] CHANG H H, YEH C H, SHEU F.A novel immunomodulatory protein from Poria cocos induces Toll-like receptor 4-dependent activation within mouse peritoneal macrophages[J].Journal of Agricultural and Food Chemistry, 2009, 57(14):6 129-6 139.
[19] 武文佳. 小麦胚芽蛋白的免疫活性评价及球蛋白免疫活性肽的结构鉴定与机理研究[D].广州:华南理工大学, 2017.
WU W J.Evaluation of immunomodulating activities in wheat germ protein, characterization and mechanism of immunomodulatory peptide in globulin[D].Guangzhou:South China University of Technology, 2017.
[20] SON H J, EO H J, PARK G H, et al.Heracleum moellendorffii root extracts exert immunostimulatory activity through TLR2/4-dependent MAPK activation in mouse macrophages, RAW264.7 cells[J].Food Science & Nutrition, 2020, 9(1):514-521.
[21] SONG Y, YAN L C, XIAO W W, et al.Enzyme-treated soy protein supplementation in low protein diet enhanced immune function of immune organs in on-growing grass carp[J].Fish & Shellfish Immunology, 2020, 106:318-331.
