Table 3.

The healthy effects of anthocyanins in mulberry.

Anti-Oxidant
Mulberry	Anthocyanins	Duration of Study	Models	Method	Effect	Reference
10 mulberry cultivars	C3G and P3G			HPLC-QTOF-MS	↓ α-glucosidase	[38]
Morus alba L.	C3G and C3R		Human hepatoma cell HepG2	HPL cytotoxicity assay C, Western blot analysis	↓ acetyl coenzyme A carboxylase activities ↑ the lipolytic enzyme expressions of PPARα and CPT1	[96]
Fresh mulberry		16-week	Male C57BL/6 mice 4 weeks old	Animal experiment with high-fat diet, Enzyme-linked immunosorbent assay (ELISA), RT-PCR	↓ MDA production ↑ SOD and GP.sub.X activities	[99]
Morus alba L.	C3G, C3R and P3G	Animal experiments: 10-week	HepG2 cells, Male db/db mice	Histology and immunohistochemistry analysis, Western blot analysis, transmission electron microscopy	↓ islet degeneration, which may be due to autophagy stimulation ↓ impaired mitochondria dysfunction ↓ excessive free radical production	[40]
Morus alba L.	C3G			Total reducing power assay (TRP assay), DPPH assay, ferric reducing antioxidant power (FRAP assay)	↓ DPPH free radical ↓ stress-induced oxidative damage	[43]
11 mulberry cultivars			Human intestinal epithelial cell line Caco-2	HPLC-TOF-MS, Determination of cellular ROS, Ccll culture	↑ ROS scavenging activity	[46]
Morus alba L.	cyanidin 3-O-(6″-O-α-rhamnopyranosyl-β-d-glucopyranoside) (C3RG), cyanidin 3-O-(6″-O-α-rhamnopyranosyl-β-d-galactopyranoside) (C3RGa), cyanidin 3-O-β-d-glucopyranoside (C3G), cyanidin 3-O-β-d-galactopyranoside (C3Ga) and cyanidin 7-O-β-d-glucopyranoside (C7G)			HPLC, ESI-MS, nuclear magnetic resonance (NMR)	↓ DPPH free radical	[104]
Morus alba L.		2-week	Male Kunming mice (18–20 g)	HPLC-PDA analysis, spectrophotography	↓ DPPH free radical and superoxide anion radicals ↑ antioxidant enzymatic activities (SOD, CAT, GSH-Px)	[105]
Anti-diabetic
Morus alba L.	C3G and C3R	5-week	Male C57BL/KsJ-db/db mice 5 weeks old	Insulin tolerance test, HPLC, the radioimmunoassay with an enzyme-linked	↑ AMPK and AS160 in skeletal muscles ↓ gluconeogenesis in the liver ↑ phosphorylated (p)-AMP-activated protein kinase (pAMPK), p-Akt substrate of 160kDa (pAS160) and plasma membrane-glucose transporter 4 (GLUT4) in skeletal muscles ↓ the levels of glucose 6-phosphatase and phosphoenolpyruvate carboxykinase in the liver	[19]
Morus alba L.	C3G, C3R, P3G and pelargonidin 3-rutinoside	6-week	Murine macrophage-like cells and rat renal tubular epithelial cells, Five-week-old male ZDF (Lepr fa/CrlCrlj) and age-matched lean rats (Lepr fa/±)	Cell culture, MTT assay, HPLC-ESI-MS/MS, histology and immunohistochemistry analysis	↓ islet degeneration and the progressive decline in insulin secretion ↓ type 2 diabetes	[106]
Mulberry	C3G		MIN6N pancreatic β-cells (derived from a mouse pancreatic islet)	Cell culture, MTT assay, immunofluorescent staining, flow cytometric and Western blot analyses	↓ intracellular reactive oxygen species, DNA fragmentation and the rate of apoptosis ↓ pancreatic β -cell apoptosis induced by high glucose conditions ↑ insulin secretion	[107]
Morus alba L.		Animal experiment: 8-week	HepG2 cells, Four-week-old male C57BL6/J genetic background (db/db) mice and their nondiabetic lean littermates (m/m)	Cell culture, histology and immunohistochemistry analysis, MTT assay, Western blot analysis, RT-PCR	In vitro: ↓ insulin resistance, ↓ PGC-1α and forkhead box protein O1 (FOXO1), enzyme activities of phosphoenolpyruvate carboxykinase (PEPCK) and glucose-6-phosphatase (G6Pase) ↑ glucose consumption, glucose uptake and glycogen content In vivo: ↓ fasting blood glucose, serum insulin, leptin, triglyceride and cholesterol levels ↑ adiponectin levels	[108]
Morus alba L.		2-week	Male Kunming mice (18–20 g)	HPLC-PDA analysis, spectrophotography	↓ fasting blood glucose, glycosylated serum Protein and anti-α-glucosidase	[105]
Morus alba L.			Human non-tumor hepatic cell line, LO2, C. elegans maintenance	Glucose consumption and uptake assays, ROS, O2−, mitochondrial membrane potential (MMP) and mitochondrial numbers assays, Western blot, RNA isolation and qPCR analysis	alleviate cellular damage and this effect is related to Nrf2 ↑ the life span of C. elegans ↓ lipid peroxidation accumulation ↑ SOD and GPx activity and PMK-1 expression	[109]
Anti-inflammatory
Morus alba L.			Peritoneal macrophages (Female BALB/c strain mice (10 weeks old) weighing 20–25 g)	HPLC, ELISA, cell culture, ELISA, MTT	↓ splenocytes’ (IFN-γ + IL-2 + IL-12)/IL-10 (Th1/Th2) cytokine secretion ratios and TNF-α/IL-10 (pro-/anti-inflammatory) cytokine secretion ratios	[67]
Fresh mulberry		16-week	Male C57BL/6 mice at 4 weeks of age	Animal experiment with high-fat diet, ELISA, RT-PCR	↓ TNFα, IL-6, iNOS and NF-κB	[99]
Morus alba L.	C3G, C3R and P3G	Animal experiments: 10-week	HepG2 cells, Male db/db mice (C57BL6/J genetic background, 4 weeks of age) and their nondiabetic lean littermates (m/m)	Histology and immunohistochemistry analysis, Western blot analysis, transmission electron microscopy	↓ the epididymal adipose mRNA expression of PPARγ, IL-6 and IL-1β ↑ the expression of SREBP-1c and C/EBP	[40]
Morus alba			Peritoneal macrophages, 6 weeks old female BALB/cByJNarl mice	Cell culture, MTT, ELISA	Mulberry juice (10–500 μg/mL): ↓ pro-inflammatory cytokines TNF-a secretions by LPS-stimulated peritoneal macrophages ↑ the secretion of anti-inflammatory cytokine IL-10 Mulberry juice (10μg/mL): ↑ TNF-a Mulberry juice (500μg/mL): ↑ IL-10	[110]
Morus alba L.		6-week	Seven-week-old male C57BL/6J mice weighing 20 g	HPLC, histology and immunohistochemistry analysis, Western blot analysis	↓ iNOS, COX2, NF-κB, TNF- a and IL-6	[111]
Anti-cancer
Morus alba L.	C3G and C3R		A549, a human lung cancer cell line obtained from ATCC (Manassas, VA, USA)	MTT assay, cell migration and invasion assays	↓ matrix matalloprotinase-2 and urokinase-plasminogen activator (u-PA) ↑ the expression of tissue inhibitor of matrix matalloprotinase-2 (TIMP-2) and plasminogen activator inhibitor (PAI)	[93]
Mulberry Fruit (Moris fructus)			A172 cells	Immunohistochemistry analysis, Western blot analysis	↓ tumor cell proliferation ↑ tumor cell apoptosis	[112]
Morus alba L.		Dextran sulfate sodium (DSS)-induced acute colitis model: 19-day; MUC2−/− mouse model: 3-month	RAW 264.7 macrophages, Six to eight-week-old BALB/c mice, and MUC2 −/− mice with colorectal cancer	Cell culture, real-time quantitative PCR, Western blot analysis, ELISA, Animal disease models and diets	↓ proinflammatory mediators and cytokines (iNOS, COX-2, IL-1β and IL-6) ↓ prevent NF-κB/p65 and MAPK/pERK signals	[113]
Morus alba L.		Animal experiments: 5-week	C57BL/6 mice, B16-F1 (a murine melanoma cell line)	Western blot analysis HPLC, MTT assay, Wound-healing assay, Boyden Chamber migration assay	↓ NF-κB transcriptional factor ↓ DNA binding activity of NF-κB to the NF-κB response element ↓ MMP-2 and MMP-9 activities	[114]
Morus alba L.			SW1736 (BRAFV600E/wt) and HTh-7 (NRASQ61R) thyroid cancer cells	MTT colorimetric assays, cell migration and invasion assays, Colony formation assay, GFP-LC3 transient transfection, flow cytometry assay, Western blot analysis	↑ apoptosis and autophagy ↓ thyroid cancer cell proliferation and Akt/mTOR signaling	[115]
Lyophilized fruit of Mulberry	C3G and C3R	Animal experiments: 49-day	AGS cell line (obtained from the Bioresource Collection and Research Center), Balb/c nude mice (male, 5 weeks old)	Western blotting analysis, HPLC	↑ intrinsic and extrinsic apoptosis through p38/p53 and p38/c-jun signaling pathways	[116]
Mulberry juice		24-h	BALB/c mice	Animal experiment	↓ the growth of Porphyromonas gingivalis, Prevotella melaninogenica	[117]
Anti-obesity
Morus alba L.			3T3-L1 preadipocyte cells (mouse embryonic fibroblast-adipose like cell line)	GC–MS, cell culture, Western blot analysis, TUNEL assay	↑ mitochondrial dysfunction, DNA Fragmentation and cell apoptosis ↓ the proliferation of 3T3-L1 preadipocyte cells	[19]
Morus alba L.	C3G and C3R		human hepatoma cell HepG2	HPL cytotoxicity assay C, Western blot analysis	↓ fatty acid synthesis ↑ fatty acid oxidation ↓ the expression of sterol regulatory element-binding protein-1 and its target molecules	[96]
Fresh mulberry		16-week	The male C57BL/6 mice with 4 weeks of age	Animal experiment with high-fat diet, Enzyme-linked immunosorbent assay (ELISA), RT-PCR	↓ serum glucose and leptin levels	[99]
Morus australis Poir	C3G, C3R and P3G	12-week	The male C57BL/6 mice with 4 weeks of age	Animal experiment with high-fat diet (HFD)	↓ insulin resistance, the size of adipocytes, lipid accumulation and leptin secretion	[118]
Morus alba L.	C3G, C3R, P3G and pelargoni-dine-3-rutinoside	12-week	6-week-old male hamsters	HPLC/ESI-MS-MS	↓ serum triacylglycerol, cholesterol, free fatty acid, the LDL/HDL ratio ↑ the hepatic peroxisome proliferator-activated receptor R and carnitine palmitoyltransferase-1 ↓ fatty acid synthase and 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase ↓ the ratio of gonadal fat and pararenal fat	[119]
Mulberry fruit	C3G, C3R		Mouse 3T3-L1 cells,	qRT-PCR, assay kit, cell culture	↓ intracellular lipid content, TG, the expression of adipogenic genes in adipocytes which may be associated with AMPK activation	[48]
Dried mulberry fruit powder obtained from M. alba L.		Animal experiments:3 months	3T3-L1 preadipocytes; female C57BL/6J mice with 8 weeks of age	LC-MS, Animal experiment with high-fat diet (HFD), histological analysis, biochemical analyses,	↓ TG, TC/HDLC, cell fat ↑ HDLC	[120]
Morus alba L.		10-week	Thirty male New Zealand white rabbits, weighing 2000–2200 g	Animal experiment with high-cholesterol diet (HCD)	↓ serum cholesterol and triglyceride and repress progression of atherosclerosis	[121]
Morus alba		12-week	Male C57BL/6 J mice (3 weeks of age)	Histological analysis, biochemical analyses, enzyme activity analyses, qRT-PCR	↓ insulin and glucose levels ↓ plasma lipid levels and lipid accumulation in the liver ↑ the activities of hepatic fatty acid b-oxidation enzymes (CPT and ACO), Ppara mRNA expression and plasma adiponectin level	[122]

↑ : increase of substance; ↓ : decrease of substance.