Table 3.

Hepatoprotective, anti-inflammatory, anti-atherosclerosis, and anti-diabetic activities of CK and its derivatives.

Material Type	ST	Model	Treatments	Major Findings	Ref.
Hepatoprotective
CK	In vivo	SVP-induced SD rats	LCK-80 mg/kg GCK + SVP MCK-160 mg/kg GCK + SVP HCK-320 mg/kg GCK + SVP once daily for 15 days	↓ hepatic index-LCK (7.6%), MCK (8.7%), and HCK (9.4%) ↓ AST, ALT, ALP, TG and ↑ALB ↑ CAT, GPx, and SOD activities and GSH level ↓ MDA level and soluble epoxide hydrolase (better with LCK) ↑ hepcidin level	[9]
CK and Rh1	In vivo	HFD-treated SD rats	CK + phospholipid; phospholipid + Rh1; phospholipid + CK+ Rh1 (3 mg/kg/day), 1 week	Treatment either alone or in combined form (CK or Rh1) ↓ γ-GT, AST, ALT, ALP, TG, CHOL, FCHOL, LDL ↑ HDL levels Anti-fibrotic effects by ↓ expressions of TIMP-1, PC-I, and PC-III Improved insulin resistance by normalizing glucose levels	[53]
	In vitro	Rat liver stellate cell line (HSC-T6)	CK, Rh1, CK+Rh1 for 6 h	↑anti-proliferative effect ↑ apoptosis in HSC-T6 CK (20.63%), Rh1(12.43%), CK+Rh1 (18%)
CK	In vivo	HFD-treated OLETF rats	CK (25 and 10 mg/kg), 12 weeks	↓ plasma glucose level and improved morphology of liver cells ↓ FAS and SREBP-1c expressions ↑ CPT-1 and PPAR-α expressions ↑ phosphorylation of AMPK	[54]
CK from GBCK25	In vivo	C57BL/6 mice	GBCK 25 with CK (400, 200, 100, 20, and 10 mg/kg) once daily, 12 weeks	↓ liver weight ↓inflammation, degree of steatosis, and ballooning degeneration ↓ ALT, TC and TG levels ↓ TNF-α, IL-1β, IL-6 levels ↓ expressions of α-SMA and TIMP-1 Reduction in hepatic lipid accumulation and ↓ MDA levels ↓ FAS, ACCα and CYP2E1 levels ↓ levels of p-JNK (reduced JNK activation)	[55]
	In vitro	Palmitic acid-treated AML12 cells LPS-treated RAW264.7 cells Kupffer cells (KCs)*	GBCK25 (4, 2, and 1 μg/mL), 24 h GBCK25 (0.5, 0.4, or 0.3 μg/mL), 24 h	↓ cellular toxicity ↓ TG, FAS, ACCα and CYP2E1 levels ↓ TNF-α, IL-1β, IL-6 in RAW264.7 and KC cells
Anti-inflammatory
CK	In vitro	LPS-stimulated RAW264.7 cells and HEK293 cells transfected with HA-AKT1, HA-Src, or HA-AKT2 for 48 h	CK (10, 5, and 2.5 μM), 24 h	No effect on the viability ↓ expressions of TNF-α, IL-1β, iNOS, and AOX1 ↓ phosphorylation of Akt1, not Akt2	[2]
BIOGF1K	In vitro	Pretreated RAW264.7 cells	BIOGF1K (200, 100, and 50 μg/mL), 1 h + LPS (1 μg/mL), 24 h	↓ NO production (67%) with BIOGF1K (200 μg/ mL) Significant scavenging of DPPH ↓ expressions of iNOS and IFN-β ↓ NF-kB activity (72%), IRF3 pathway (63%) Inhibited IKK and TBK1 phosphorylation	[56]
BIOGF1K	In vitro	Pretreated RAW264.7 cells	BIOGF1K (30, 20, and 10 μg/mL), 30 min + LPS (1 μg/mL), 24 h	Dose-dependent ↓ of NO and iNOS and COX-2 expressions AP-1 signaling pathway inhibited by blocking MAPKs and MAPKKs	[57]
BSA-CK NPs	In vitro	Pretreated RAW 264.7 cells	BSA-CK NPs and CK (20,15, 10, 5, and 1 µM), 1 h + LPS (1 mg/mL)	↓ NO production by BSA-CK NPs (10 µM) compared with CK	[39]
SPIONs-CK	In vitro	Pretreated RAW 264.7 cells	SPIONs-CK and CK (100, 10, and 1 μg/mL), 24 h + LPS (1 μg/mL) Antioxidant-1 to 250 μg/mL	↓ NO production by CK and SPION-CK and inhibited iNOS production by 47.9% (CK) and 45.8% (SPION-CK) (at 10 μg/mL) ↓ ROS production by SPIONs-CK and CK Inhibition of DPPH was higher for SPIONs-CK (72%) compared to CK (21.1%) at (250 μg/mL)	[58]
CK	In vivo	C57BL/6 mice	CK (20 mg/kg), 30 h	↑expression of SGLT1 gene and glucose uptake mediated by SGLT1	[59]
	In vitro	Caco-2 cells	CK (1, 0.1, 0.01, and 0.001 µM), 12, 24, 36, and 48 h	↑ SGLT1 protein level dose-dependent ↑ SGLT1 protein level time-dependent 1.70 times (24 h) to 2.01 times (48 h) ↑ glucose uptake activity by ↑ SGLT1 expressions
CK	In vivo	Xylene-induced Kunming mice with ear swelling	CK (224, 112, 56, 28, 14, and 7 mg/kg) every day, 5 days	CK displayed a dose-dependent inhibitory effect At 224 mg/kg- maximum (93.9%) inhibition	[60]
		Carrageenan-induced paw oedema SD rats	CK (160, 80, 40, 20, 10, and 5 mg/kg), orally every day, 5 days	Pain threshold induced by heat not effected ↑ rat inflammatory pain threshold significantly ↓ PGE2 level in the paw tissue, not in the gastric mucosa. ↓ COX-2 level in the gastric mucosa and paw tissue Activities COX-1 and -2 not effected
CK	In vivo	CIA-induced DBA/1 mice	CK (224, 56, and 14 mg/kg) per day, 21 days	Significant ↓ in arthritis global assessment and swollen joint count ↑ number of naïve T-cells and ↓ activated T-cells and DCs percentage Inhibited migration and priming of DCs ↓ expressions of CD80, CD86, MHC II, and CCL 21 levels (lymph nodes)	[61]
CK	In vivo	CIA-induced DBA/1 OlaHsd mice	CK (100 μl) once a day (20, 10, and 5 mg/kg/day), 6 weeks (Preventive effect), 4 weeks (Therapeutic effect)	↓ arthritis scores, ↓ serum anti-CII IgG, IFN-γ, and IL-2 ↑ IL-4 levels Non-significant ↓ TNF-α and IL-17 levels ↓ RANKL/OPG and MMP-3/TIMP-1 ratios	[62]
CK	In vivo	Adjuvant-induced arthritis	CK (160, 40, and 10 mg/kg), once daily, 15 days	Significant ↓ in global assessment scores and swollen joint counts ↓ spleen index and hyperplasia of lymph nodes ↓ memory B cells in the spleen ↓ expressions of CD40L (T cells) and CD40 (B cells)	[63]
CK	In vivo	CIA-induced DBA/1 mice	CK (112 mg/kg/day), 24 days	Recovered body weight and ↓ arthritis symptoms, spleen index Inhibited viability and proliferation of lymphocytes ↓ IL-1β, IL-17 and TNF-α and ↑ IL-10 ↓ M1 macrophages and ↑ M2 macrophages; prevented phagocytosis ↑ Gαs expression and inhibited β-arrestin2, NF-κB, TLR4, and Gαi	[64]
CK	In vitro	H2O2-stimulated MC3T3-E1 cells	CK (0.01-10 μM) with or without H2O2, 48 h	CK formed hydrogen bonds with IKK ↑ ALP activity, Col-I expressions, and mineralization ↓ ROS and NO production, IL-1β expression	[65]
GNP-CK-CopA3	In vitro	LPS-activated RAW264.7	GNP-CK-CopA3 (10-100 µg/mL), 1 h + LPS (1 µg/mL), 24 h	NO production was inhibited (at 20 and 40 µg/mL) ROS production inhibited-40.4% (20 µg/mL) and 65.05% (40 µg/mL) ↓ levels of TNF-α, iNOS, COX-2, IL-6, and IL-1β Inhibited NF- κB and MAPK signaling pathways	[66]
Anti-atherosclerosis
CK	In vivo	ApoE-/- C57BL/6 Peritoneal macrophages from apoE-/- C57BL/6	CK (9, 3, and 1 mg/kg) one dose per day, 8 weeks. ox-LDL (100 µg/mL) + CK (30, 10, and 3.3 µM)	↓ atherosclerotic plaques (55%) by activating RCT pathway ↓ IL-6, IL-1β, and TNF-α levels ↓ cleaved IL-1β, caspase-1, NLRP3, and NF-kB P65 ↓ inflammasome activity in mice and macrophages ↓ cholesterol ester (10 μM 46.21% and 30 μM 60.24%)	[67]
CK and its derivatives	In vitro	RAW264.7 cells	CK and CK derivatives (30, 10 µM)	Structure 1 ↓ cholesteryl ester contents in foam cells compared to CK ↑ ABCA1 mRNA expression Structure 1 (319%) compared to CK (151%) Structure 1 significantly activated LXRα compared to CK No effect on LXRβ activation	[68]
CK	In vitro	HUVECs	Pretreated with CK (2.5, 1.25, and 0.625 mM), 12 h + ox-LDL (80 mg/mL), 24 h	↓ expressions of IL-6, MCP-1, TNF-α, VCAM1, and ICAM-1 ↓ expression of caspase3, cleaved caspase-3 and cytochrome c and LDH release Reversed mitochondrial membrane depolarization ↑ Bcl2/Bax	[69]
Anti-diabetic
CK	In vivo	HFD fed ICR mice	Injected with STZ (100 mg/kg BW) after 4 weeks + CK (30 mg/kg), 4 weeks	↓ blood glucose levels, improve glucose tolerance ↓ PGC-1α expressions and inhibited PEPCK, G6Pase expressions Improved AMPK phosphorylation	[70]
	In vitro	HepG2 cells	CK (8, 4, and 2 μM), 24 h	Dose-dependent inhibition of hepatic glucose production ↓ PEPCK protein level and ↑ AMPK phosphorylation
CK and Rb1	In vivo	Epididymal adipose tissue from ICR mice	Glucose treatment (high concentration), 24 h + CK (10 μM) and Rb1 (10 μM)	↓ROS production and ERS ↓ phosphorylation of PERK and IRE1a ↓ activation of NLRP3 inflammasome and ↓ IL-1β, IL-6 production ↓ IRS-1 phosphorylation at a serine residue ↑ IRS-1 phosphorylation at tyrosine residue ↑ PI3K activity and Akt phosphorylation	[71]
CD-CK conjugate	In vivo	Alloxan-induced diabetic zebrafish model	CK and CD-CK (15, 10, 7.5, 5, 2.5, 1, 0.5, 0.1, and 0.05 μM), 2 days	Good recovery of pancreatic islets by CD-CK compared to CK CD-CK showed less toxic (LC50 = 20.68 μM) than CK (LC50 = 14.24 μM)	[72]
CK conjugate with beta-cyclodextrin	In vivo	HFD-induced C57BL/6 mice	CK (40, 20, and 10 mg/kg/day), 8 weeks	↑ body weight (6th week) ↓ fasting glucose, BUN, creatinine, and urine protein ↓ ROS production and Nox1, Nox4 expressions↓ expressions of NLRP3, Caspase-1, ASC, IL-1β, TNF-α, and IL-18 CK treatment reduced the activation of the p38 MAPK signaling pathway	[73]
	In vitro	High glucose-treated HBZY-1 cells	CK (50, 40, 20, and 10 μM), 48 h	↓ proliferation of HBZY-1 cells ↓ NLRP3, Caspase-1, and ASC levels

* ex vivo, ST, study type; CK, compound K, SVP, sodium valproate; SD, Sprague-Dawley; LCK, low CK; GCK, ginsenoside CK; MCK, middle CK, HCK, high CK; AST, aspartate transaminase; ALT, alanine aminotransferase; ALP, alkaline phosphatase; TG, triglyceride; ALB, albumin; CAT, catalase; SOD, superoxide dismutase; GPx, glutathione peroxidase; GSH, glutathione; MDA, malondialdehyde; HFD, High fat diet; γ-GT, gamma-glutamyl trans peptidase; CHOL, total cholesterol; FCHOL, free cholesterol; LDL, low density lipoprotein; HDL, high density lipoprotein; TIMP-1, tissue inhibitors of metalloproteinase-1; OLETF, otsuka long-evans tokushima fatty; FAS, fatty acid synthase; SREBP-1c, sterol regulatory element-binding protein-1c; CPT-1, carnitine palmitoyltransferase-1; PPAR-α, peroxisome proliferator-activated receptor-alpha; AMPK, 5′ AMP-activated protein kinase; TC, total cholesterol; TNF-α, tumor necrosis factor alpha; IL, interleukin; α-SMA, alpha smooth muscle actin; ACCα acetyl CoA carboxylase alpha; CYP2E1, cytochrome P450 2E1; p-JNK, phospho c-Jun N-terminal kinase; LPS, lipopolysaccharide; iNOS, inducible nitric oxide synthase; AOX1, aldehyde oxidase 1; Akt, protein kinase B; BIOGF1K, CK and F1; NO, nitric oxide; DPPH, 2,2-diphenyl-1-picrylhydrazyl; IFN-β, interferon-beta; NF-kB, nuclear factor-kB; IRF3, interferon regulatory factor 3; IKK, inhibitor of kB kinase; TBK1, TANK-binding kinase 1; COX-2, cyclooxygenase-2; AP-1 (also known as c-jun), activator protein-1; MAPKs, mitogen-activated protein kinases; MAPKKs, MAPK kinases; BSA, bovine serum albumin; NP, nanoparticles; SPIONs, superparamagnetic iron oxide nanoparticles; ROS, reactive oxygen species; SGLT, sodium-glucose linked transporter or sodium-dependent glucose cotransporters; PGE2, prostaglandin E2; CIA, collagen-induced arthritis; DCs, dendritic cells; CD, cluster of differentiation; MHC, major histocompatibility complex; CCL21, chemokine (C-C motif) ligand 21; CII IgG, type II collagen immunoglobulin G; RANKL, receptor activator of nuclear factor-κB ligand; OPG, osteoprotegerin; MMP, matrix metalloproteinase; TLR4, Toll-Like receptor 4; Gαis, G(i,s) protein subunit alpha; Col-I, type I collagen; GNPs, gold nanoparticles; apoE, apolipoprotein E; ox-LDL, oxidized low density lipoprotein; RCT, reverse cholesterol transport; NLRP3, NOD-like receptor protein-3; ABCA1, ATP-binding cassette transporter A1; LXR, liver X receptor; HUVECs, human umbilical vein endothelial cells; MCP-1, monocyte chemoattractant protein-1; VCAM-1, vascular cell adhesion molecule-1; ICAM-1, intercellular adhesion molecule-1; LDH, lactate dehydrogenase; Bcl2, B-cell lymphoma-2; Bax, B-cell lymphoma 2 (BCL-2)-associated X protein; ICR, imprinting control region; STZ, streptozotocin; PGC-1α, proliferator-activated receptor-γ coactivator-1 alpha; PEPCK, phosphoenolpyruvate carboxykinase; G6Pase, glucose-6-phosphatase; ERS, endoplasmic reticulum stress; IRE1, inositol-requiring enzyme 1; PERK, protein kinase-like ER kinase; IRS-1, insulin receptor substrates -1; PI3K, phosphatidylinositol 3 kinase; BUN, blood urea nitrogen; Nox, NADPH oxidase; ASC, apoptosis-associated speck-like protein containing a CARD.