Table 2.

Anti-inflammatory properties of phytosterols

Molecules	Origins	Experimental methods	Key findings	Refs.
β-sitosterol	Ranunculus sceleratus L	MTT assay 5-lipoxygenase (5-LOX) activity Oxazolone-induced mouse ear oedema	Reduced oedema (in vivo) No effect on arachidonate pathway enzymes implicated in the inflammatory process (in vitro)	[63]
–	Purchased	Human aortic endothelial cells (HAECs) Monocyte (U937 cells) Attachment in tumor necrosis factor-α (TNF-α)-stimulated HAECs MTT assay Cytokine quantitation: ELISA	Inhibited intracellular adhesion molecule 1 (ICAM-1) and vascular cell adhesion molecule 1 (VCAM-1) expression in TNF-α Inhibited U937 cell binding to TNF-α Attenuated nuclear factor-κB (NF-κB) p65 phosphorylation	[90]
–	Nyctanthes arbortristis	Doses: 5, 10, and 20 mg/kg, i.p Wistar rats Hind paw oedema	Inhibited paw oedema	[110]
–	Trachelospermum jasminoides	Lipopolysaccharide (LPS)-stimulated RAW264.7 murine macrophages	Inhibited the activities of interleukin-6 (IL-6) of stimulated macrophages Reduced TNF-α and IL-1β secretion	[111]
–	Purchased	2,4,6-trinitrobenzene sulfonic acid (TNBS)-induced colitis in mice C57BL/6 mice Immunoblotting ELISA measurement	Decreased myeloperoxidase (MPO) activity (in vivo) Attenuated NF-κB Inhibited inflammatory enzymes Inhibited pro-inflammatory cytokines	[72]
–	Esenbeckialeiocarpa	Carrageenan-induced inflammation in the mouse air pouch model Calcium uptake measurement MPO and adenosine-deaminase (ADA) activity quantification	Increased calcium uptake in activated neutrophils Inhibited ADA and MPO activity Inhibited TNF-α and IL-1β levels	[73]
–	Purchased	Mice with colitis after being fed a high fat diet (HFD) Intestinal mouse macrophages stimulated with lipopolysaccharides (LPS) ELISA measurement Immune blot analysis	Ameliorated HFD-induced colitis through inhibition of LPS binding to toll-like receptor 4 (TLR4) in the NF-κB pathway	[71]
–	Justicia gendarussa burm. F	Carrageenan-induced rat paw oedema model Calculated the percentage inhibition of oedema	Induced potent anti-inflammatory effect by secreting prostaglandin (69.43%), serotin and bradykinin (52.25%), and histamine (30.07%)	[62]
–	Purchased	26 C57BL/6 J male mice were fed a high fat western-style diet (HFWSD) and subsequently developed colitis through the induction of dextran sulfate sodium (DSS)	Reduced the severity of colitis Suppressed NF-κB activation Decreased colonic inflammation score Decreased COX-2 expression	[69]
–	Oxalis corniculata	Doses: 5, 10, and 20 mg/kg, i.p Rat paw oedema model	After 120 min, the administration of a dosage of 20 mg/kg resulted in: The inhibition of rat paw edema, reducing it to a volume of 0.32 ± 0.06 mL	[61]
–	Moringa oleifera	Doses: 7.5 − 30 μM HaCaT cells (keratinocytes) J774A.1 cells (macrophages)	Secretion of inflammatory factors (IL-1β, IL-6, TNF-α, and ROS) from both keratinocytes and macrophages was effectively suppressed Inhibited caspase-1 activation Partially inhibited NF-κB in macrophages	[101]
–	Purchased	Wistar male rats ELISA measurement RT-PCR analysis	Inhibiting inflammatory reactions, particularly TNF-α	[112]
–	Purchased	LPS-exposed BV2 murine microglial cells CCK-8 assay ELISA measurement RT-PCR analysis Western blot analysis	Reduced inflammatory mediator [IL-6, COX-2, TNF-α, and inducible nitric oxide (iNOS)] expression Inhibited NF-κB, ERK, and p38 pathway activation	[80]
–	Purchased	Induction of arthritis via complete freund’s adjuvant Swiss Wistar rats Poly-arthritic index ELISA measurement qRT-PCR analysis	Reduced paw edema and arthritic index Reduced cytokines Reduced the level of COX-2, prostaglandin E2 (PGE2), NF-κB, and vascular endothelial growth factor (VEGF)	[64]
–	Purchased	Dose: 1 mg/kg Puncture (CLP)-induced septic rats ELISA measurement RT-qPCR analysis	Decreased cytokine levels	[97]
–	Not reported	Dose: 150 mg/kg, p.o Female Wistar rats ELISA measurement	Down-regulated IL-6 and TNF-α levels Up-regulated endothelial nitric oxide synthase (eNOS) level	[85]
–	Purchased	RAW264.7 cells were stimulated with LPS, while C57BL/6 J mice developed acute lung injury as a result of LPS induction ELISA measurement RT-PCR analysis Western blot analysis	Suppressed the mRNA levels of IL-6, TNF-α, and IL-1β (in vitro) Inhibited the activation of TLR4/NF-κB pathway activation	[108]
–	Purchased	A zebrafish larvae (Danio rerio) model of inflammation Neutrophil migration assay	Inhibited neutrophil migration to the lateral line Reduced inflammatory gene (myd88 and il-8) expressions	[78]
Ergosterol	Sarcodon aspratus (Berk.) S. Ito	Murine macrophages (RAW264.7) HT29 colorectal adenocarcinoma cells	Inhibited TNF-α secretion Inhibited IL-1α/β expression Suppressed JNK, MAPK p38, and ERK phosphorylation	[98]
–	Purchased	LPS-induced inflammation in RAW264.7 macrophages MTT assay Flow cytometry analysis Western blotting analysis	Down-regulated proteins associated with the NF-κB cascade Inhibited TNF-α production Inhibited COX-2 expression	[99]
–	Cordyceps militaris	BV2 microglia cells	Reduced nitric oxide (NO) in LPS triggered BV2 cells	[87]
–	Scleroderma Polyrhizum Pers.,	Doses: 25 and 50 mg/kg LPS-induced acute lung injury	Inhibited inflammatory cells Inhibited pro-inflammatory cytokines (TNF-α and IL-6) Inhibited NF-κB, COX-2, and iNOS activation	[104]
–	Purchased	Chronic obstructive pulmonary disease (in vivo) ELISA measurement Western Blot analysis	Inhibited cytokines in lung and serum Inhibited expression of JAK3/STAT3/NF-κB pathway (in vivo)	[95]
–	Ganoderma sinense	NO production in RAW264.7 macrophages	Exhibited anti-inflammatory activities against NO production IC50 = 17.7–32.4 μM	[86]
–	Purchased	CS-induced chronic obstructive pulmonary disease (COPD) model (in vivo and in vitro) 6HBE cells and Balb/c mice ELISA measurement Western blot analysis NF-κB/p65 activity assay	Inhibited CS-induced o inflammation by inhibiting NF-κB/p65 activation	[21]
–	Anvillea garcinii	5-LOX inhibitory potential	Displayed moderate 5-LOX inhibitory activity (IC50 = 3.06 − 3.57 mM)	[107]
–	Purchased	Diabetic nephropathy-induced inflammation (in vivo) ELISA measurement RT-PCR analysis Western blot analysis	Decreased inflammatory cytokine (MCP-1, TNF-α, and IL-6) levels Decreased NF-κB signaling pathway	[102]
–	Cryptoporus volvatus	LLC-PK1 cells RT-PCR analysis Western blot analysis Immunofluorescence assay Flow cytometry assay	Decreased the mRNA levels of cytokines (PKR, Mx1, IFN-β, IFN-α)	[92]
–	Sclerotinia Sclerotiorum	Croton oil-induced ear edema quantification of MPO enzyme	Inhibited ear edema, via inhibition of COX pathway activity Inhibited neutrophil recruitment	[77]
–	Purchased	TNF-α-induced HT-22 hippocampal cell injury Western blot analysis RT-PCR analysis	Attenuated TNF-α toxicity on HT-22 cells Induced the overexpression of the EGR-1 transcription factor	[113]
–	Antrodia camphorata	LPS-induced microglial activation and neuro-inflammatory reactions (in vivo and in vitro) BV2 and HMC3 microglial cells Male ICR mice Western blot analysis Immunohistochemistry RT-PCR analysis	Inhibited protein kinase B (AKT), NF-κB, and mitogen-activated protein kinase (MAPK) signaling pathways (in vitro) Decreased the levels of NF-κB phosphorylation and cytokines (in vivo)	[26]
Stigmasterol	Mondia whytei	Doses: 7.5, 15, 30, and 100 mg/kg Dimethylbenzene-induced ear edema test Adult Swiss albino mice	Inhibited ear edema Induced anti-inflammatory activity (50.34% at 30 mg/kg)	[66]
–	Purchased	Human umbilical vein endothelial cells MTT assay Western blot analysis RT-PCR analysis Immunohistochemistry	Down-regulated TNF-α No effect on other cytokines (IL-6 and CXCL-8)	[96]
–	Purchased	Colitis in 26 C57BL/6 J male mice fed a HFWSD	Reduced the severity of colitis Suppressed NF-κB activation Decreased colonic inflammation score Decreased COX-2 expression	[69]
–	Purchased	Irritant dermatitis	Controlled inflammatory characteristics (neutrophilia and ear skin edema) Suppressed pro-inflammatory cytokine expression Reduced TNF-α serum levels	[65]
–	Purchased	Collagen-induced arthritis (CIA)-induced inflammation in rats ELISA measurement qPCR analysis	Inhibited TNF-α/IL-6/IL-1β/iNOS/COX-2 expression Augmented anti-inflammatory cytokine IL-10 expression Down-regulated NF-kBp65/p38MAPK expression Inhibited p-IKB-α activation	[22]
–	Purchased	Peritonitis Paw edema	Reduced paw edema Decreased leukocyte infiltration Prevented paw licking Presented anti-inflammatory effects	[67]
–	Purchased	IL-1β-induced inflammation in primary rat articular chondrocytes MTT assay qRT-PCR analysis Western blot analysis	Reduced IL-6 and iNOS expression Stigmasterol + mesenchymal stem cells-condition medium (MSC-CM) normalized pro-inflammatory/pro-catabolic responses and inhibited NF-κB activation	[23]
–	Petiveria alliacea	Pro-inflammatory mediator release by egg albumin protein Molecular Docking Studies against TNF-α and COX-2	At 6.5 kcal/mol: Better binding energy against TNF-α At 9.7 kcal/mol: Identified as hit molecule against COX-2 enzyme	[114]
–	Purchased	APPswe/PS1dE9 mice Simulated BV2 cells with Aβ42 oligomers ELISA measurement CCK8 assay Flow cytometry assay Western blot analysis	Reduced microglia activation Reduced pro-inflammatory cytokine levels Protected BV2 cells via AMPK/NLRP3 and AMPK/NF-κB signaling	[24]
–	Not reported	RAW264.7 macrophages Splenocytes MTT assay qRT-PCR analysis Western blot analysis ELISA measurement	Increased IL-2 and IFN-γ levels in RAW264.7 cells Up-regulated NF-κB activation in RAW264.7 cells Increased TNF-α secretion levels in splenocytes	[34]
Fucosterol	Undaria pinnatifida	LPS-induced RAW264.7 macrophages Western blot analysis qRT-PCR analysis	Inhibited iNOS, TNF-α, and IL-6 expression Suppressed p38 MAPK and NF-κB pathways	[81]
–	Eisenia bicyclis	LPS-induced RAW264.7 macrophages MTT assay	Suppressed COX-2 and iNOS expression Suppressed NF-κB pathway	[79]
–	Hizikia fusiformis	Atopic dermatitis-like lesions (in vivo) Mouse macrophage cells RAW264.7 macrophages Rat basophilic leukemia RBL-2H3 cells ELISA measurement	Decreased LPS-induced production of NO (in vitro) Decreased DNCB-stimulated serum immunoglobulin E (IgE) levels Inhibited TNF-α and IL-4 levels Increased IFN-γ secretion	[88]
–	Purchased	Acute lung injury (in vivo) Alveolar macrophages (in vitro) MTT assay ELISA measurement Western blot analysis	Attenuated cytokine production (in vivo) Inhibited NF-κB activation and cytokine production (in vitro)	[100]
–	Hizikia fusiformis	Hypoxia damages to keratinocytes (HaCaT)	Demonstrated a dose-dependent suppression of inflammation Inhibited the expression of IL-1β, IL-6, and TNF-α in HaCaT cells Suppressed HIF1-α accumulation	[103]
–	Purchased	β-amyloid (Aβ)- and LPS-induced microglial cells ELISA measurement RT-PCR analysis	Inhibited IL-1β, IL-6, TNF-α, PGE2, and NO production	[89]
–	Sargassum binderi	Particulate matter (PM)-induced inflammation in A459 human lung epithelial cells Western blot analysis	Suppressed COX-2 and PGE2 Inhibited TNF-α and IL-6 Suppressed MAPK and NF-κB pathways	[93]
–	Padina boryana	PM-induced inflammation in RAW264.7 macrophages MTT assay Western blot analysis qRT-PCR analysis	Down-regulated dose-dependently the production of inflammatory mediators (PGE2, COX-2, and iNOS) and pro-inflammatory cytokines (TNF-α, IL-6, and IL-1β) Suppressed NF-κB and MAPK pathways	[52]
–	Sargassum horneri	Human dermal fibroblast (HDF) Western blot analysis RT-PCR analysis	Down-regulated inflammatory mediators Regulated Nrf2/HO-1 signaling Regulated NF-κB and MAPK signalling	[25]