A Comprehensive Review on Phytochemistry and Pharmacological Activities of Clinacanthus nutans (Burm.f.) Lindau

Table 7

Pharmacological activities of C. nutans.

(a) Pharmacological activity: antivenom

Experiment design; (Experiment model; Venom; Assay; Test subject)	Extract	Plant part	Plant Source	Extract dose; Route of administration	Result	Reference
in vitro; Heterometrus laoticus scorpion venom; Pre-incubated extract with 0.2 μg/μL venom-Cell lytic test incubated for 30 min; CEFs	Water	Leaf	Thailand	0.706, 0.406 mg/mL	Extract at 0.706 mg/mL give 46.51% of efficiency but the cytotoxic of extract is questionable	[65]
in vitro; Apis mellifera Linn. bee venom; Pre-incubated extract with 0.6 μg/μL venom-Cell lytic test incubated for 30 min; CEFs	Water, 90% and 50% ethanol	Leaf	Thailand	0.706, 0.406 mg/mL	Ineffective	[66]
in vivo; Laticauda colubrine snake venom; Mice and Mongrel dog	Water; Maceration	Leaf	Sarawak; Malaysia	6 mg/kg per mouse (i.p), 20 mg/kg per dog (i.v)	Ineffective	[7]
in vivo; Naja naja siamensis snake venom; Isolated rat phrenic-nerve diaphragm preparations; Mice	Water	Leaf	Thailand	p.o, i.p	Ineffective	[5]
in vivo; Snake venom; Mice	Water	Leaf	Thailand	-	Reduce mortality rate from 100% to 63 ± 3.34%	[67]
in vivo; Snake venom; Mice	95% alcohol	Leaf	Thailand	2000 mg/kg; i.v, i.p, p.o	Ineffective	[67]

CEFs: Chick embryo fibroblasts.

(b) Pharmacological activity: analgesic/antinociceptive

Experiment design; (Experiment model; Assay; Test subject)	Extract	Plant part	Plant Source	Extract dose; Route of administration; positive control	Result	Reference
in vivo; Acetic acid-induced writhing test; Mice	Water, methanol, chloroform, n-butanol	Leaf	Thailand	Phenylbutazone (100 mg/kg)	Effective (n-butanol extract at 90 mg/kg)	[68, 69]
in vivo; Acetic acid-induced abdominal writhing test-Pre-treatment 1 h before test; ICR mice (adult male, 25–30 g)	100% Methanol; Oven dry; Soaking	Leaf	Kuala Lumpur Malaysia	100, 250, and 500 mg/kg; ASA (100 mg/kg)	ED50: 279.3 mg/kg	[19]
in vivo; Formalin-induced paw licking test-Pre-treatment 1 h before test; Sprague Dawley rat (adult male 150–180 g)	100% Methanol; Oven dry; Soaking	Leaf	Kuala Lumpur Malaysia	100, 250, and 500 mg/kg; ASA (positive, 100 mg/kg); morphine (5 mg/kg)	Early phase: ED50: >500 mg/kg; Late phase: ED50: 227.7 mg/kg	[19]
in vivo; Hot plate test at 50°C- Pre-treatment 1 h before test; ICR mice (adult male; 25–30 g)	100% Methanol; Oven dry; Soaking	Leaf	Kuala Lumpur Malaysia	100, 250, and 500 mg/kg; morphine (5 mg/kg)	500 mg/kg significant delay response at the interval of 60 to 210 min	[19]
in vivo; Hot glass jar at 72°C water bath; Albino mice (either sex, 40–50 g)	95% Ethanol; Maceration	Leaf	Thailand	5 g/kg	Ineffective	[92]
in vivo; Hot water plate test; Mice	n-Butanol	Leaf	Thailand	p.o, i.p; morphine	Ineffective	[68, 69]

(c) Pharmacological activity: anti-inflammatory

Experiment design; (Experiment model; Assay; Test subject)	Extract	Plant part	Plant Source	Extract dose; Route of administration; Positive control	Result	Reference
in vitro; fMLP induced elastase release- Pre-treatment; Human neutrophils	Methanol; Air dry; Percolation	Whole plant	Thailand	0.01–100 μg/mL; Indomethacin (1–100 μg/mL)	IC50: 186.8 ± 20.5 μg/mL; elicited a weak, but significant inhibition of human neutrophil elastase release (100 nM)	[70]
in vitro; fMLP/CB induce elastase release; Human neutrophils	80% ethanol, ethyl acetate, n-hexane; Air dry; Soaking	Aerial	Taichung; Taiwan	10 μg/mL	80% ethanol showed highest inhibition: 68.33 ± 5.49%	[52]
in vitro; fMLP induced neutrophil superoxide anion generation (pre-incubation for 10 min)	Methanol; Air dry; Percolation	Whole	Thailand	0.01–100 μg/mL; Indomethacin	IC50: 23.4 ± 3.1 μg/mL	[70]
in vitro; Superoxide anion generation assay; Human neutrophils	80% ethanol, ethyl acetate, n-hexane; Air dry; Soaking	Aerial	Taichung; Taiwan	10 μg/mL	80% ethanol showed highest inhibition: 28.52 ± 2.55%	[52]
in vitro; fMLP induced neutrophil myeloperoxidase (MPO) production	Methanol; Air dry; Percolation	Whole	Thailand	0.01–100 μg/mL; Indomethacin	IC50: 219.5 ± 25.7 μg/mL; Inhibition via reduced neutrophil migration	[70]
in vitro; Immunoblotting-LPS induced TLR-4 inflammatory proteins; Protein lysate from macrophage	Polar; Soaking	Leaf, stem	Pahang Malaysia	20 μg/mL	Significantly reduced the LPS induced phosphorylation of p65, p38, ERK1/2, JNK1/2, IRF3	[62]
in vitro; LPS induced TLR-4 assay; HEK-Blue-hTLR4 cells	Polar, nonpolar; Soaking	Leaf, stem	Pahang Malaysia	100 μg/mL	IC50: 21.3 ± 5.0 (leaf polar) to 29.4 ± 9.0 (leaf non-polar) μg/mL	[62]
in vitro; LPS induced cytokine production assay-1 h pre-treatment; murine macrophages RAW 264.7 cell	Polar, nonpolar; Soaking	Leaf, stem	Pahang Malaysia	100 μg/mL	Polar leaf (p < 0.05) inhibited TNF-α, IFN-γ, IL-1β, IL-6, IL12p40, IL-17 production	[62]
in vitro; LPS induced NO- 1 h pre-treatment- Griess assay; Murine macrophages RAW 264.7 cell	Polar, nonpolar; Soaking	Leaf, stem	Pahang Malaysia	100 μg/mL	IC50: 18.9 ± 3.6 (leaf polar) to 43.1 ± 4.7 (leaf non-polar) μg/mL	[62]
in vivo; Acetic acid induced vascular permeability model; Mice	Water, methanol, chloroform, n-butanol	Leaf	Thailand	Indomethacin (4 mg/kg)	Effective, n-butanol at 540 mg/kg as potent as indomethacin	[68, 69]
in vivo; EPP induced rat ear oedema model- pre-treatment; Sprague Dawley rats (male, 40–60 g)	Methanol; Air dry; Percolation	Whole plant	Thailand	3, 6, 9 mg/20 μL acetone per ear; Apply topically; Indomethacin (2 mg/20 μL)	At dose 9 mg EPP/ear: 79% oedema inhibition at 15 min, 44.4% MPO reduction after 120 min of induction	[70]
in vivo; Carrageenan induced paw oedema model-1 h pre-treatment; Sprague Dawley rats (male, 100–120 g)	Methanol; Air dry; Percolation	Whole plant	Thailand	50, 100, 200 mg/kg; p.o; Indomethacin (20 mg/kg)	200 mg/kg of extract inhibit 59% of oedema at 3 h	[70]
in vivo; Carrageenan induced paw oedema; Albino mice (40–50 g)	95% Ethanol; Maceration	Leaf	Thailand	5000 mg/kg; p.o	17.73% at 3 h and 36.47% at 6 h of oedema inhibition	[92]
in vivo; Carrageenan induced paw oedema model; Mice	n-Butanol	Leaf	Thailand	p.o; acetylsalicylic acid (100 mg/kg)	Effective, n-butanol at 270 mg/kg as potent as ASA	[68, 69]
in vivo; Granuloma pouch model; Wistar rats (male)	Aqueous ethanol cream	Leaf	Thailand	125 mg cream/rat; Apply topically; Prednisolone (0.25%)	Inhibit 48.3% granuloma formation	[71]
in vivo; Granuloma pouch model; Wistar rats (male)	95% ethanol cream	Leaf	Thailand	125 mg cream/rat; Apply topically; Prednisolone (0.25%)	Inhibit 50.1% granuloma formation	[71]
in vivo; Granuloma pouch model-1 h pre-treatment; Mice	n-Butanol cream	Leaf	Thailand	270 and 540 mg/kg; Apply topically	Ineffective	[68, 69]
in vivo; Granuloma pouch model; Wistar rats (male)	Cold cream	Leaf	Thailand	125 mg cream/rat; Apply topically; Prednisolone (0.25%)	Inhibit 50.98% granuloma formation	[71]

(d) Pharmacological activity: immunomodulating effect

Experiment design; (Experiment model; Assay; Incubation period; Test subject; Extract dose; Positive control)	Extract	Plant part	Plant Source	Result	Reference
in vitro; with and without fMLP induced Chemotaxis and Chemokinesis; 45 min; Human neutrophils; 0.1–100 μg/mL; Indomethacin (0.01–100 μg/mL, IC50: 56.3 ± 3.5 ng/ml)	Methanol; Air dry; Percolation	Whole plant	Thailand	Chemotaxis: With fMLP induction, IC50: 2.7 ± 0.6 μg/mL, Without fMLP, IC50: >100 μg/mL Chemokinesis: With fMLP induction, IC50: 5.5 ± 0.6 μg/mL, Without fMLP, IC50: 5.0 ± 0.5 μg/mL	[70]
in vitro; Apoptosis assessment via morphological, flow cytometry-Annexin V binding; 20 h; Human neutrophils; 10–500 μg/mL; Dexamethasone (1 μM, inhibited 54.5% neutrophil apoptosis)	Methanol; Air dry; Percolation	Whole plant	Thailand	No significant effect on neutrophil apoptosis	[70]
in vitro; ConA & LPS-induced IL-10 & TNF-α expression-Real time PCR analysis; 16 h; PBMC; 1.56 mg/mL	Methanol; Oven dry; Maceration	Leaf, Stem	Chiang Mai Thailand	Reduce IL-10 mRNA expression, no modulating effect on TNF-α mRNA expression	[72]
in vitro; IFN-γ expression Splenocyte from ovalbumin-primed BALB/c mice	80% Ethanol; Air dry; Soaking	Aerial	Taichung Taiwan	0.1 μg/mL: upregulation of IFN-γ.; 100 μg/mL: down-regulation of IFN-γ	[52]
in vitro; HaCaT; IFN-γ/TNF-α-induced apoptosis- MTT assay; 1 and 100 μg/mL	Ethanol; Maceration	Leaf	Thailand	Significantly suppressed keratinocytes apoptosis	[63]
in vitro; Lymphocyte proliferation assay-^3H- thymidine-radioactivity; 72 h; HBMC; 0.5–5000 μg/mL	Ethanol; Soxhlet	Leaf	Thailand	0.5–5 μg/mL cause proliferation increase, 2.5, 5 mg/mL cause proliferation decrease	[73]
in vitro; NK activity-K562 as target cell-cytotoxicity assay; 72 h; HBMC; 0.5–5000 μg/mL	Ethanol; Soxhlet	Leaf	Thailand	1 and 5 mg/mL-cause NK activity decrease	[73]
in vitro; IL-2 production-ELISA; 72 h; HBMC; 0.5–5000 μg/mL	Ethanol; Soxhlet	Leaf	Thailand	undetectable IL-2	[73]
in vitro; IL-4 production-ELISA; 72 h; HBMC; 0.5–5000 μg/mL	Ethanol; Soxhlet	Leaf	Thailand	2.5 and 5 mg/mL-increase IL4 production	[73]
in vitro; Lymphocyte subpopulation-flow cytometry assay; 72 h; HBMC; 0.5–5000 μg/mL	Ethanol; Soxhlet	Leaf	Thailand	no change in the percentages of CD3^+, CD4^+, CD8^+, CD16^+/CD56^+ and CD19^+ cells	[73]

(e) Pharmacological activity: neuroprotective and neuromodulating function

Experiment design; (Experiment model; Assay; Incubation period; Test subject)	Extract	Plant part	Plant Source	Extract dose; Route of administration; Positive control	Result	Reference
in vitro; OGD–reoxygenation and hypoxic neuronal death, Cell viability assay; Mouse primary cortical neurons, cerebral astrocytes, cerebral endothelial cells	80% Ethanol; Soaking	Leaf	Singapore	5 μg/mL	Extract suppressed post-hypoxic HDACs activation and reduce OGD-caused neuronal death	[75]
in vitro; OGD–reoxygenation; 24 h Human SH-SY5Y neuroblastoma cells	80% Ethanol; Soaking	Leaf	Singapore	100 μg/mL	Extract modulated cPLA2 expression induction in SH-SY5Y cells by HDAC inhibitors, MS-275, MC-1568, TSA and inhibited HAT activity.	[74]
in vitro; OGD–reoxygenation; 12 h and 24 h; Mouse primary cortical neurons	80% Ethanol; Soaking	Leaf	Singapore	1.6, 6.25 μg/mL	Extract inhibited levels of cPLA2 mRNA expression in primary cortical neurons subjected to 0.5 h OGD injury	[74]
in vitro; OGD; CCK-8; up to 12 h; neutron pre-treated with extract 1 h before OGD; Mouse primary cortical neurons from E15.5 Balb/c mouse embryos	80% Ethanol; Soaking	Leaf	Malaysia	6.25 μg/mL	Extract treated neurons showed significant increment in cell viability	[76]
in vitro; OGD–reoxygenation; CCK-8, MMP, apoptosis analysis, transient transfection and chromatin reporter assay; 0.5 h OGD+ 4–24 h Reoxygenation; Neurons treated with extract (1 h before, on the onset or after OGD-reoxygenation); Mouse primary cortical neurons from E15.5 Balb/c mouse embryos	80% Ethanol; Soaking	Leaf	Malaysia	0.075–20 μg/mL	Neurons treated with extract before, at the onset or after OGD showed increment in dose dependent manner but protective effect of extract was lesser when applied after OGD.; Neurons treated with 6.25 μg/mL extract at the onset of OGD has reduced MMP breakdown, apoptotic death, and pro-apoptotic (caspase-3, PARP-1) and has higher anti-apoptotic (14-3-3ε, p-Bad, Bcl-2) markers.; Neurons treated with 2.5–10 μg/mL extract at the onset of OGD suggested a dose dependent increase effect in PPAR-γ mRNA level.	[76]
in vitro; Anti-spasmotic effect on acetycholine induced rat’s bladder tissue contraction; Isolated tissues of rat’s bladder	Ethanol	Leaf, stem	Malaysia	Mebeverine hydrochloride and detrusitol	Stem at 120 mg/mL: 79.77%; Leaf at 100 mg/mL: 26.62%; Detrusitol at 200 μg/mL: 89.86%, Mebeverine at 10 mg/mL: 80.74% of contraction inhibition	[93]
in vivo; Ellman assay-Acetylcholinesterase activity in brain, liver, kidney, heart; Pretreatment for 14 days (daily treatment (once)); Balb/C mice (male, 25 ± 5 g)	Methanol, Air dry, Maceration	Leaf	Seremban Malaysia	250, 500, 1000 mg/kg bw	All doses of extracts caused the higher acetylcholinesterase activity in liver, kidney, and heart compared to control group while they did not caused any difference in brain	[77]
in vivo; MCA occlusion stroke model; Reperfusion infarct volume, behaviour assessment, MRI; extract treated 1 h before, immediate or 3–24 h after 30 min MCA occlusion or 1 d after 15 min MCA acclusion; Long- Evans rats (male, 7-8 week-old)	80% Ethanol; Soaking	Leaf	Malaysia	10–60 pg (i.c.v., after 30 min MCA occlusion); 24 mg/kg body wt (i.p, 1 h before or 3–24 h after 30 min or 24 h after 15 min MCA occlusion)	I.C.V. treatment of 40 pg extract reduced apoptotic neuronal death, showed a maximal decrease of cerebral infarct volume at 1 day reperfusion. PPAR-γ, C/EBPβ mRNA levels, anti- apoptotic 14-3-3ε, p-Bad, and Bcl-2 levels were upregulated. Pro-apoptotic cleaved PARP-1 and cleaved caspase 3 protein levels were downregulated. Extract improved the functional outcomes based on the behavior observation at day 7 after ischemia (elevated body swing test and Bederson’s postural reflex task) and at day 14 after ischemia (better ladder rung walking test).; Extract via i.p treatment also significantly reduced infarct volume except 24 h after 30 min MCA occlusion. Behavior test conducted also showed better functional outcomes and smaller infarct volume via MRI.	[76]

(f) Pharmacological activity: antidiabetic and α-glucosidase inhibition activity

Experiment design; (Experiment model; Assay; Test subject)	Test sample	Plant part	Plant Source	Extract dose; Route of administration; Positive control	Result	Reference
in vitro; α-glucosidase inhibition assay	Hot water; Oven dry	-	Malaysia	50 mg/mL	88.2% of inhibition	[37]
in vitro; α-glucosidase inhibition assay	Methanol; Oven dry; Sonication	Leaf, Stem	Seremban Malaysia	5000 μg/mL (in stock)	13.57 (leaf), 17.67 (stem) % of inhibition	[61]
in vitro; α-glucosidase inhibition assay	70% Ethanol; Oven, air, freeze dry; Sonication, soaking	Leaf, stem	Seremban Malaysia	5000 μg/mL (in stock); Quercetin	Lowest: 5.31% (Leaf-Freeze dry-Soaking), Highest: 41.70% (Leaf-Oven dry-sonication)	[4]
in vitro; α-glucosidase inhibition assay	CO2-Soxhlet, Maceration	Stem	Seremban Malaysia	5000 μg/mL (in stock); Quercetin	CO2-Soxhlet: 95.79% of inhibition; Maceration: 58.23% of inhibition	[28]
in vivo; Alloxan induced model- daily treatment for 9 days; Swiss webster mice (male)	Hot water; Sun dry	Leaf	Bandung Indonesia	50, 100, and 150 mg/kg BW; p.o; Oral glibenclamide	150 mg/kg significantly lower blood glucose serum level from 442 ± 149 mg/dl (day 0) to  mg/dl (day 9)	[25]
in vivo; High fat and high cholesterol diet (HFHC) induced insulin resistance-7 weeks periods-daily treatment of extract; Sprague Dawleys rat (male, 200–250 g)	Water, 80% methanol; Sonication	Leaf	Seremban Malaysia	500, 250 or 125 mg/kg/day/rat; p.o.; Simvastatin	C. nutans attenuated the metabolic effects and transcriptional changes induced by the HFHC diet	[39]
in vivo; Glucose solution (p.o, 2 g/kg bw); Swiss webster mice (male)	Insoluble ethyl acetate fraction from ethanol	Leaf	Bandung Indonesia	100 mg/kg bw; p.o	Decreased 18.4% blood glucose serum level in 3 h	[25]

(g) Pharmacological activity: antioxidant

Test sample	Plant part	Plant Source	Extract dose; Positive control	Result	Reference
(1) Type of assay: in vitro; DPPH scavenging assay
Hot water; Oven dry	-	Malaysia	1 to 16 mg/mL	16 mg/mL: ~60% inhibition	[37]
Hot water; Unfermented and fermented; Microwave-oven dry, freeze dry; Infusion for 1, 2, 5, 10, 15, and 20 min	Leaf	Sabah Malaysia	0.1, 0.2, 0.4, 0.6, 0.8, 1.0 mg/mL	Unfermented possessed higher DPPH inhibition than fermented tea	[36]
Hot water, water, 80% methanol, methanol, ethyl acetate, hexane; Freeze dry; Sonication	Leaf, stem	Seremban Malaysia	Trolox	6.12% (hexane leaf) to 55.12% (80% methanol leaf) inhibition	[42]
Water, methanol, chloroform; Oven dry; Soaking	Leaf	Serdang Malaysia	12.5, 25, 50 and 100 μg/mL; Trolox	Chloroform > methanol > water; 864.11 ± 73.49 to 7852.63 ± 449.90 (chloroform) μg Teq/g extract	[32]
Methanol; Sun dry	Aerial	Vietnam	0, 20, 40, 60, 80, 100 μg/mL	IC50: 114.50 μg/mL	[49]
Methanol; Air dry; Percolation	Whole	Thailand	1–400 μg/mL; Trolox	IC50: 240.1 ± 15.3 mg/mL	[70]
Methanol; Air dry; Maceration	Leaf	Seremban Malaysia	0.25 to 10 mg/mL; Quercetin	IC50: 1.33 ± 0.0001 mg/mL	[24]
Methanol; Oven dry; Soaking	Leaf, stem	Seremban Malaysia	5000 μg/mL (stock); Quercetin	Leaf: IC50: 1126.63 μg/mL; Stem: IC50: 1548.89 μg/mL	[61]
Methanol; 1, 6 and 12 months old; Freeze dry	Bud, leaf	Serdang Malaysia	BHT, caffeic acid, α-tocopherol	IC50: 64.6 (1 year old buds) to 112.1 (1 year old leaf) μg/mL	[41]
Methanol, Petroleum ether, Ethyl acetate; Oven dry; Soaking	Leaf, stem	Taiping Malaysia	0.2 to 10.0 mg/mL	4 mg/mL: leaf petroleum ether give 82% of DPPH inhibition	[88]
Methanol, ethyl acetate, chloroform, hexane	Whole	Jelebu Malaysia	1000 μg/mL; BHT	50.50 ± 0.03% (Hexane) to 70.96 ± 0.03% (chloroform) inhibition	[53]
98% methanol, ethyl acetate, chloroform, hexane; Sun dry; Soaking	Stem	Vietnam	Vitamin C	Ineffective	[47]
50% ethanol; Soaking	Leaf	Thailand	1–300 μg/mL; Ascorbic acid	:  μg/mL	[94]
70% ethanol; Freeze dry; Sonication	Leaf	Perak; Malaysia	200–1000 μg/mL; Green tea	not determined	[64]
70% ethanol; Freeze dry, oven dry, air dry; Sonication, soaking	Leaf, stem	Seremban Malaysia	5000 μg/mL (stock); Quercetin	15.44 ± 2.21% (Stem-freeze dry-soaking) to 44.31 ± 3.16% (Leaf-oven dry-sonication) inhibition	[4]
80% Ethanol; Young, old; Storage duration (1, 2, 3, and 4 days)	Leaf, stem	Malaysia	-	Maturity: 31.24% (matured stem) to 112.12% (young leaf); Storage duration: 31.88% (4 days) to 101.85% (1 day) inhibition	[38]
Ethanol; Maceration	Aerial	Bangkok; Thailand	1 mg/mL	Ineffective	[78]
Ethyl acetate	Leaf	Indonesia	BHT	: 178.40 mg/L	[95]
CO2-Soxhlet, Maceration	Stem	Seremban Malaysia	5000 μg/mL (stock); Quercetin	CO2-Soxhlet: 98.92% inhibition; Maceration: 63.00% inhibition	[28]
(2) Type of assay: in vitro; FRAP assay
Hot water; Unfermented and fermented; Microwave-oven dry, freeze dry; Infusion for 1, 2, 5, 10, 15, and 20 min	Leaf	Sabah Malaysia	0.1, 0.2, 0.4, 0.6, 0.8, 1.0 mg/mL	250.70 ± 49 (fermented-microwave oven dry-20 min infusion) to 438.80 ± 94 (unfermented-freeze dry-10 min infusion) mg/L	[36]
Hot water, water, 80% methanol, methanol, ethyl acetate,; Hexane; Freeze dry; Sonication	Leaf, stem	Seremban Malaysia.	Gallic acid	~45 (hexane stem) to ~148 mg (hot water leaf) GAE/g extract	[42]
Methanol, ethyl acetate, chloroform, hexane	Whole	Jelebu Malaysia	1000 μg/mL; Ascorbic acid	hexane and ethyl acetate (not determined) to 56.49 ± 0.05% (methanol)	[53]
Methanol; 1, 6 and 12 months old; Freeze dry	Bud, leaf	Serdang Malaysia	BHT, caffeic acid, Vitamin C	209 μM of Fe (II)/g (1-month-old leaves) to 488 μM of Fe (II)/g (6-month-old buds).	[41]
70% Ethanol; Freeze dry; Sonication	Leaf	Perak; Malaysia	200–1000 μg/mL; Green tea	very low absorbance (ineffective)	[64]
50% Ethanol; Soaking	Leaf	Thailand	1–100 μg/mL; Ascorbic acid	17 mg ascorbate/g extract	[94]
(3) Type of assay: in vitro; Hydrogen peroxide scavenging activity
Water, methanol, chloroform; Oven dry; Soaking	Leaf	Serdang Malaysia	12.5, 25, 50 and 100 μg/mL; Quercetin	Methanol: highest radical scavenging: ~34% at 100 μg/mL	[32]
(4) Type of assay: in vitro; Metal chelating activity
Hot water; Oven dry	-	Malaysia	1, 2, 5, 10 mg/mL	10 mg/mL: ~90% inhibition	[37]
(5) Type of assay: in vitro; Nitric oxide scavenging assay
Hot water; Oven dry	-	Malaysia	1, 5, 10 mg/mL	10 mg/mL: ~90% inhibition	[37]
Water; methanol, chloroform; Oven dry; Soaking	Leaf	Serdang Malaysia	12.5, 25, 50 and 100 μg/mL	Water: 32.33% at 100 μg/mL	[32]
(6) Type of assay: in vitro; ABTS cation radical scavenging assay
Hot water; unfermented and fermented; microwave-oven dry, freeze dry; hot boiling water (100°C); infusion for 1, 2, 5, 10, 15, and 20 min	Leaf	Sabah Malaysia	0.1, 0.2, 0.4, 0.6, 0.8, 1.0 mg/mL	50.36 ± 4.07 (fermented-microwave oven dry-10 min infusion) to 74.03 ± 2.26 (unfermented-freeze dry-5 min infusion) mg AEAC/L	[36]
Hot water, water, 80% methanol, methanol, ethyl acetate,; hexane; Freeze dry; Sonication	Leaf, stem	Seremban Malaysia	Trolox	18 (hexane leaf) to 65 (80% methanol-leaf) mg TE/g extract	[39]
(7) Type of assay: in vitro; Galvinoxyl radical scavenging activity
Water; methanol, chloroform; Oven dry; Soaking	Leaf	Serdang Malaysia	12.5, 25, 50 and 100 μg/mL; Trolox	Chloroform > methanol > aqueous; Chloroform: 12248.82 μg Teq/g extract	[32]
(8) Type of assay: in vitro; Superoxide radical scavenging activity
Methanol; Air dry; Percolation	Whole	Thailand	Direct EPR scavenging effect	showed significant direct scavenging activity when the incubation time extended to 60 min	[70]
70% ethanol; Freeze dry; Sonication	Leaf	Perak; Malaysia	Green tea; by NBT method	3651.5 U/g	[64]
(9) Type of assay: in vitro; Phorbol 12-myristate 13-acetate (PMA) induced peroxide production in rat macrophages
50% Ethanol; Soaking	Leaf	Thailand	10, 30, 100 and 300 μg/mL; Tiron	Fluorescent intensity: 58.72 ± 5.52 (30 μg/mL), 51.92 ± 8.49 (100 μg/mL), 53.50 ± 6.17 (300 μg/mL)	[94]
(10) Type of assay: in vitro; Protective effect against peroxyl radicals initiator (AAPH)-induced oxidative hemolysis
50% Ethanol; Soaking	Leaf	Thailand	200–1000 μg/mL; Ascorbic acid	IC50: 359.38 ± 14.02 μg/mL	[94]
(11) Type of assay: in vivo; Hyperlipidemia-associated oxidative stress model
Water, 80% methanol; Freeze dry; Sonication	Leaf, stem	Seremban Malaysia	125, 250, 500 mg/kg; Sprague Dawley rats (male, 200–250 g); daily treatment for 49 days; p.o; Simvastatin	Both leaf extracts reduce oxidative stress through increasing serum antioxidant enzymes activity and upregulating the expression of hepatic antioxidant genes	[42]

(h) Pharmacological activity: antimicrobial-antiviral

Experiment design; (Experiment model; Virus strain; Assay; Test subject; Route of administration/Prescription)	Extract	Plant part	Plant Source	Extract dose; Positive control	Result	Reference
Type of virus: Varicella zoster virus (VZV)
in vitro; VZV Kawaguchi strain; DNA hybridization technique, plaque reduction assay-pre, post, direct; WI-38 HEL cells	Organic; Soxhlet	Leaf	Thailand	Acyclovir; IC50: 107 μM (pre), 5 μM (post), 30 μM (direct)	Most effective: Direct inactivation; IC50: 1 : 2000 (pre), 1 : 6000 (post), >1 : 18000 (direct); IC50: 1 : 2000 (pre), 1 : 4800 (post), 1 : 9600 (direct)	[80]
Clinical; Herpes zoster (shingles); Double-blinded, randomized trials; 60 patients; Apply topically 5 times daily for 7 to 14 days	5% extract cream	-	Thailand	Placebo	Lesion crusting within 3 days: C. nutans (89.3%), Placebo (0%); Lesion healing within 7 days: C. nutans (100%), Placebo (100%); Pain scores: C. nutans better than placebo group	[8]
Clinical; Herpes zoster (shingles); Randomized trials; 48 patients; Apply topically 5 times daily for 5 days	5% extract cream	-	Thailand	Placebo, Acyclovir	Lesion crusting within 3 days: C. nutans (81.3%), Acyclovir (27.2%), Placebo (0%); Lesion healing within 7 days: C. nutans (81.30), Acyclovir (18.20%), Placebo (0%)	[82]
Clinical; Herpes zoster (shingles); Double-blinded, block randomization; 120 patients; Apply topically 3 times daily from 1 to 26 days	120 patients	-	Thailand	Placebo	Lesion crusting within 3 days: C. nutans (27.5%), Placebo (8.6%); Lesion healing: C. nutans (14 days) Placebo (18 days); Symptom reduction: C. nutans better thanplacebo groups	[81]
Type of virus: Herpes simplex virus (HSV)
in vitro; HSV-1-KOS strain; Plaque reduction assay-post; Vero cell	Methanol, dichloromethane, n-hexane; Soxhlet	Leaf	Thailand	12.5–100 μg/mL; Acyclovir (IC50: 0.09 μg/mL)	IC50: 32.05 ± 3.63 (n-hexane) to 64.93 ± 7.00 (methanol) μg/mL	[83]
in vitro; HSV-1F strain; Plaque reduction assay-pre, post; Vero cell	Ethyl acetate; Oven dry; Soxhlet	Leaf	Thailand	19 μg/mL; Acyclovir (5 μg/mL, post), Dextran sulfate (2 μg/mL, pre)	Pre: IC50: 7.6 μg/mL; significantly reduce plaques; Post: ineffective	[79]
in vitro; HSV-1 strain; Plaque reduction assay-pre, post-incubated for 48 h; Vero cell	Chloroform; Soxhlet	Leaf	Thailand	Acyclovir (IC50: 0.64 ± 0.07 μg/mL)	Pre: less than 50% inhibition of plaque formation; Post: IC50: 115.00 μg/mL	[56]
in vitro; HSV-2-strain G, 5 clinical HSV-2 isolates; Plaque reduction assay (post), Yield reduction assay (post), Inactivation kinetics (direct, 4 h); Vero cell	Methanol; Air dry; Percolation	Whole	Thailand	2500 μg/mL	Ineffective	[96]
in vitro; HSV-2-Baylor 186; Plaque reduction assay-post; Vero cell	Methanol, dichloromethane, n-hexane; Soxhlet	Leaf	Thailand	12.5-100 μg/mL; Acyclovir (IC50: 0.43 μg/mL)	IC50: 65.13 ± 2.22 (methanol) to 72.62 ± 12.60 (n-hexane) μg/mL	[83]
in vitro; HSV-2 standard strain; Plaque reduction assay-pre, post, direct; BHK cell	Ethanol; (A2, C1, C2, C3, C4, C5 fraction)	Leaf	Thailand	-	C2, C3, C4: 1 : 2400 dilution cause 100% plaque inhibition, through extracellularly pathway	[97]
in vitro; HSV-2 strain; Plaque reduction assay-pre, post-incubated for 96 h; Vero cell	Chloroform; Soxhlet	Leaf	Thailand	Acyclovir (IC50: 0.80 μg/mL)	Post: IC50: 140.00 ± 3.00 μg/mL	[56]
Clinical; HSV-2-Herpes genitalis; Sequential randomisation; 77 patients; Apply topically 4 times daily for 6 days	5% extract cream	-	Thailand	Placebo, Acyclovir (Zovirax)	Lesion crusting within 3 days: C. nutans (98.6%), Acyclovir (80.8%), Placebo (12.5%); Lesion healing within 7 days: C. nutans (88.9%), Acyclovir (84.6%), Placebo (20.8%); C. nutans has no sticky, burning, stinging pain and side effects.	[82, 84]
Clinical; HSV-2-Herpes genitalis; Randomized trials; 163 patients; Apply topically 4 times daily for 6 days	5% extract cream	-	Thailand	Placebo, Acyclovir	The lesion crusting and healing speed were significantly better in the C. nutans and acyclovir treated groups compared to the placebo groups. No side effect observed C. nutans treated group.	[85]
Type of virus: Fish pathogenic virus
in vitro; IHNV, OMV, IPNV strain; Plaque reduction assay-pre, post, direct; CHSE-214 cells	Ethanol; Soxhlet	-	Thailand	100 μg/mL (pre, post), 500 μg/mL (direct)	Direct: 100% (IHNV and OMV), 0% (IPNV) of plaque reduction; Pre: 31% (IHNV), 54% (OMV), 74% (IPNV) of plaque reduction; Post: 25% (IHNV), 48% (OMV), 3% (IPNV) of plaque reduction	[98]
Type of virus: Crustaceans (shrimp and prawn) infectious virus
in vivo; YRV-RNA virus; Anti-viral test-direct; cultured black tiger shrimp	Ethanol; Soxhlet	Leaf	Thailand	0.1 to 10 mg/mL	Minimum inhibition: 1 μg/mL	[87]
in vivo; YRV-RNA virus; Protective efficacy assay-14 days observation; daily twice treatment for 7 days-pre; cultured black tiger shrimp; p.o	Ethanol; Soxhlet	Leaf	Thailand	0, 0.1, 1 and 10 g/kg of pellet	1 g/kg of feed exhibited best protective efficacy with 57.6%	[87]
Type of virus: Mosquito-borne virus
in vitro; DENV-2 strain 16681; Western blot assay, ECL detection kit-post; Naïve Huh-7 cells	80% Ethanol; Air dry; Soaking	Aerial	Taichung Taiwan	Ribavirin	IC50: 31.04 μg/mL	[52]
Type of virus: Poultry and bird contagious virus
in vitro; NDV- La Sota strain; Hemagglutination test-pre, post; CEFs	Water, Ethanol; Soxhlet	-	Thailand	31.25 g/mL (final)	Ineffective	[86]

(i) Pharmacological activity: Anti-microbial-anti-bacterial

Experiment design; (Experiment model; Bacteria strain; Assay)	Extract	Plant part	Plant Source	Extract dose; Positive control	Result	Reference
in vitro; A. hydrophila; Agar dilution assay	Ethanol; Soxhlet	-	Thailand	0 to 10.0 mg/mL	MIC: >10 mg/mL	[99]
in vitro; B. cereus; Disc diffusion assay	70% Methanol; Air dry; Soxhlet	Leaf	Perak; Malaysia	25, 50 and 100 mg/mL; Ciprofloxacin	At 100 mg/mL:  mm	[22]
in vitro; B. cereus; Microdilution assay	Methanol; Maceration	Leaf	Seremban Malaysia	0.1 to 12.5 mg/mL; Erythromycin, chloramphenicol	MIC: >12.5 mg/mL	[6]
in vitro; B. cereus; Microdilution assay	Ethyl acetate and its fraction; Oven dry; Soaking	Leaf, stem	Taiping Malaysia	0.08 to 5 mg/mL (final); Ampicillin	MIC: Ethyl acetate: 6.31 mg/mL,; F7: 1.39 mg/mL	[88]
in vitro; B. subtilis; Light mediated disk diffusion assay	95% Ethanol; Soaking	Leaf	Thailand	5 mg/mL; Gentamycin	Ineffective	[89]
in vitro; E. coli; Microdilution assay	Hot water; Oven dry	-	Malaysia	0.4 to 50.0 mg/mL (final); Ampicillin	MIC: >50 mg/mL	[43]
in vitro; E. coli; Disc diffusion assay	70% Methanol; Air dry; Soxhlet	Leaf	Perak	25, 50 and 100 mg/ml; Ciprofloxacin	At 100 mg/ml:  mm	[22]
in vitro; E. coli; Microdilution assay	Methanol; Maceration	Leaf	Seremban Malaysia	0.1 to 12.5 mg/mL; Erythromycin, chloramphenicol	MIC: 12.5 mg/mL	[6]
in vitro; E. coli; Microdilution assay	Ethyl acetate and its fraction; Oven dry; Soaking	Leaf, stem	Taiping Malaysia	0.08 to 5 mg/mL (final); Ampicillin	MIC: Ethyl acetate: >100 mg/mL,; F7: 1.39 mg/mL	[88]
in vitro; E. coli DC10; Light mediated disk diffusion assay	95% Ethanol; Soaking	Leaf	Thailand	5 mg/mL; Gentamycin	Ineffective	[89]
in vitro; E. coli (wild); Light mediated disk diffusion assay	95% Ethanol; Soaking	Leaf	Thailand	5 mg/mL; Gentamycin	Ineffective	[89]
in vitro; M. luteus; Microdilution assay	Hot water; Oven dry	-	Malaysia	0.4 to 50.0 mg/mL (final); Ampicillin	MIC: >50 mg/mL	[43]
in vitro; MRSA; Disc diffusion assay, microdilution assay	Ethanol; Oven dry; Maceration	-	Thailand	5 mg/mL	No detected inhibition zone, MIC and MBC: >5 mg/mL	[100]
in vitro; MSSA K147; Light mediated disk diffusion assay	95% Ethanol; Soaking	Leaf	Thailand	5 mg/mL; Gentamycin	Ineffective	[89]
in vitro; N. gonorrhoeae & 11 clinical isolates; Disc diffusion	Methanol; Maceration	Leaf	Thailand	44 mg/mL (final); Norfloxacin	Ineffective	[101]
in vitro; P. acnes; Microdilution assay	Methanol; Maceration	Leaf	Seremban Malaysia	0.1 to 12.5 mg/mL; Erythromycin, chloramphenicol	MIC: >12.5 mg/mL	[6]
in vitro; P. acnes; Disc diffusion, microdilution assay	Ethanol; Maceration	Aerial	Thailand	Mangostin	MIC and MBC: >5 mg/mL	[102]
in vitro; P. aeruginosa; Microdilution assay	Hot water; Oven dry	-	Malaysia	0.4 to 50.0 mg/mL (final); Ampicillin	MIC: >50 mg/mL	[43]
in vitro; P. aeruginosa; Disc diffusion assay	70% Methanol; Air dry; Soxhlet	Leaf	Perak	25, 50 and 100 mg/ml; Ciprofloxacin	At 100 mg/ml:  mm	[22]
in vitro; P. aeruginosa 187 (wild); Light mediated disk diffusion assay	95% Ethanol; Soaking	Leaf	Thailand	5 mg/mL; Gentamycin	Ineffective	[89]
in vitro; S. aureus; Microdilution assay	Hot water; Oven dry	-	Malaysia	0.4 to 50.0 mg/mL (final); Ampicillin	MIC: >50 mg/mL	[43]
in vitro; S. aureus; Disc diffusion assay	70% Methanol; Air dry; Soxhlet	Leaf	Perak	25, 50 and 100 mg/ml; Ciprofloxacin	At 100 mg/mL:  mm	[22]
in vitro; S. aureus; Microdilution assay	Methanol; Maceration	Leaf	Seremban Malaysia	0.1 to 12.5 mg/mL; Erythromycin, chloramphenicol	MIC: 12.5 mg/mL	[6]
in vitro; S. aureus; Disc diffusion assay, microdilution assay	Methanol; Sun dry; Maceration	Aerial	Vietnam	125–1000 mg/mL; Erythromycin (10–50 μg/μL)	16.67 mm inhibition zone, MIC: 62.5 mg/mL	[49]
in vitro; S. aureus; Disc diffusion assay, microdilution assay	Ethanol; Oven dry; Maceration	-	Thailand	5 mg/mL	No detected inhibition zone, MIC: 5 mg/mL, MBC: >5 mg/mL	[100]
in vitro; S. enterica serovar Paratyphi C; Disc diffusion assay	Distilled water, 70% ethanol, absolute ethanol, chloroform	Leaf	Malaysia	-	Showed inhibition zone	[26]
in vitro; S. enterica serovar Paratyphi B; Disc diffusion assay	Distilled water, 70% ethanol, absolute ethanol, chloroform	Leaf	Malaysia	-	Chloroform: a larger inhibition zone compare to other solvents.	[26]
in vitro; S. enterica serovar Typhi; Disc diffusion assay	Distilled water, 70% ethanol, absolute ethanol, chloroform	Leaf	Malaysia	-	Least inhibition zone	[26]
in vitro; S. enterica serovar Typhimurium; Disc diffusion assay	Distilled water, 70% ethanol, absolute ethanol, chloroform	Leaf	Malaysia	-	Showed inhibition zone	[26]
in vitro; S. enterica Typhimurium; Disc diffusion, microdilution assay	Methanol; Sun dry; Maceration	Aerial	Vietnam	125–1000 mg/mL; Erythromycin (10–50 μg/μL)	15.67 mm inhibition zone, MIC: 125 mg/mL	[49]
in vitro; S. enterica Typhimurium; Microdilution assay	Ethyl acetate and its fraction; Oven dry; Soaking	Leaf, stem	Taiping Malaysia	0.08 to 5 mg/mL (final); Ampicillin	MIC: Ethyl acetate: >100 mg/mL,; F7: 1.39 mg/mL	[88]
in vitro; S. enterica serovar Weltevreden; Disc diffusion assay	Distilled water, 70% ethanol, absolute ethanol, chloroform	Leaf	Malaysia	-	Showed inhibition zone	[26]
in vitro; S. epidermidis; Microdilution assay	Methanol; Maceration	Leaf	Seremban Malaysia	0.1 to 12.5 mg/mL; Erythromycin, chloramphenicol	MIC: >12.5 mg/mL	[6]
in vitro; S. epidermidis; Disc diffusion, microdilution assays	Ethanol; Maceration	Aerial	Thailand	Mangostin	MIC and MBC: >5 mg/mL	[102]
in vitro; S. epidermidis; Disc diffusion, microdilution assays	Ethanol; Oven dry; Maceration	-	Thailand	5 mg/mL	No detected inhibition zone, MIC and MBC: 5 mg/mL	[100]
in vitro; Streptococcus sp.; Agar dilution assay	Ethanol; Soxhlet	-	Thailand	0 to 10.0 mg/mL	MIC: >10 mg/mL	[99]
in vitro; V. harveyi; Agar dilution assay	Ethanol; Soxhlet	-	Thailand	0 to 10.0 mg/mL	MIC: >10 mg/mL	[99]
in vitro; V. paraliaemolyticlls; Agar dilution assay	Ethanol; Soxhlet	-	Thailand	0 to 10.0 mg/mL	MIC: >10 mg/mL	[99]

(j) Pharmacological activity: antimicrobial-antifungal

Experiment design; (Experiment model; Fungal strain; Assay)	Extract	Plant part	Plant Source	Extract dose; Positive control	Result	Reference
in vitro; A. fumigatus; Light mediated disk diffusion assay	95% Ethanol; Soaking	Leaf	Thailand	5 mg/mL; Nystatin	Ineffective	[89]
in vitro; C. albicans; Light mediated disk diffusion assay	95% Ethanol; Soaking	Leaf	Thailand	5 mg/mL; Nystatin	Ineffective	[89]
in vitro; C. albicans; Microdilution assay	Ethyl acetate & its fraction; Oven dry; Soaking	Leaf, stem	Taiping Malaysia	0.08 to 5 mg/mL (final); Amphotericin B	Ethyl acetate: 6.31 mg/mL,; F7: MIC: 1.39 mg/mL	[88]

(k) Pharmacological activity: anticancer

Experiment design; (Experiment model; Cancer cell(s); Incubation period; Assay; Test subject; Route of administration)	Extract	Plant part	Plant Source	Extract dose; Positive control	Result	Reference
in vitro; Mutated Salmonella typhimurium (T98 and T100) without metabolic activation; 6 days; Mutagenicity assay-Ames test	Water; Air dry; Maceration	Leaf	Penang Malaysia	500 μg/well	Non-mutagenic activity in S. typhimurium histidine auxotrophs	[103]
in vitro; D24, MM418C1, MCF7, BT474 cancer cells; 24 h and 72 h; CCK-8 assay	Hot water, cold water, methanol, ethanol, dichloromethane	Leaf	Seremban Malaysia	2 mg/mL	D24 cell: cold water at 72 h caused 42.9% cell death (EC50: 1.63 mg/mL); MM418C1 and MCF7 cells: hot and cold water showed cytotoxicity; BT474 cells: extract ineffective	[18]
in vitro; K562, HCT 116 cancer cells; MTT cytotoxicity assay	Water, 50% methanol, 100% methanol, 50% ethanol, 100% ethanol	Leaf	Malaysia	100 and 200 μg/mL	Ineffective	[104]
in vitro; HepG2, IMR-32, NCI-H23, SNU-1, LS-174T, K-562, HeLa, Raji cancer cells; 72 h; MTT cytotoxicity assay	Water, ethanol, chloroform; Oven dry; Soaking	Leaf	Serdang Malaysia	3.125 to 100 μg/mL	Most effective: chloroform with IC50: 47.70 μg/mL (K562), 47.31 μg/mL (Raji)	[32]
in vitro; Cultured Saos-2 human steosarcoma cells; HIF activity, MTT cytotoxicity assay	Methanol	Leaf	Malaysia	125–2000 μg/mL	Ineffective	[105]
in vitro; HeLa cancer cells; 72 h; MTT cytotoxicity assay	Methanol; Freeze dry	Leaf, buds	Serdang Malaysia	10, 20, 40, 80, 160, 320 μg/mL; Tamoxifen	Most effective: 6-month bud with IC50: 56.8 μg/mL	[41]
in vitro; D24 melanoma cells; 24 h and 72 h, CCK-8 assay	Methanol; Air dry; Soaking with shaker	Leaf	Malaysia, Vietnam, Thailand	0–2 mg/mL (stock)	Chiang Dao Thailand extract has highest cytotoxicity: EC50: 0.95 mg/mL (24 h), 0.77 mg/mL (72 h)	[106]
in vitro; HepG2, MCF-7, NCI-H460, HeLa cancer cells; 48 h; Sulforhodamine B colorimetric assay	Sub-fraction F-III from Methanol extract	Stem	Vietnam	Camptothecin	Effective, IC50: 36.80 (HepG2), 57.36 (NCI-H460), 66.57 (MCF-7), 91.08 (Hela) μg/mL	[47]
in vitro; HepG2 cell; 24 h; MTT cytotoxicity assay	Methanol, ethyl acetate, hexane, chloroform fraction	-	Malaysia	0, 6.25, 12.5, 25, 50, 100 μg/mL (stock)	IC50: 43.93 (methanol), 55.61 (chloroform), 62.06 (ethyl acetate), 68.38 (hexane) μg/mL	[35]
in vitro; MCF-7, HeLa cells, 72 h; MTT cytotoxicity assay	Methanol, ethyl acetate; Freeze dry; Soaking	Root	Sabah Malaysia	10–50 μg/mL; Camptothecin (0.35 μg/mL)	MCF-7 cell: IC50: 35.0 μg/mL (methanol), 30.0 (ethyl acetate) μg/mL; HeLa cell: ~25% of inhibition for both extracts	[31]
in vitro; HeLa, K-562 cells; 24 h and 72 h; MTT cytotoxicity assay	Methanol, ethyl acetate, petroleum ether; Oven dry; Soaking	Leaf, stem	Taiping Malaysia	0.2–10.0 mg/mL	Petroleum ether leaf extract showed the strongest cytotoxic activity after 72 h, CC50: 18.0 μg/mL (HeLa), 20.0 μg/mL (K-562)	[88]
in vivo; MMS induced Allium cepa chromosome assay-post and suppressive treatment; Allium cepa	Water, methanol; oven dry	Leaf	Penang; Malaysia	100, 200, 400, 800 mg/kg	Extract has repairing and anti-mutagenic effect	[90]
in vivo; Hepatocarcinoma tumor-bearing mice-daily treatment for 10 days; ICR mice (with normal T/B cells, 6–8 weeks, 18–22 g); Gastric probe	30% Ethanol fraction	Aerial	Seremban; Malaysia	3 and 10 mg/kg; Fluorouracil (20 mg/kg)	8.2% (3 mg/kg) and 58.6% (10 mg/kg) of tumor size and weight reduction, higher reduction rate than positive control (37.1%)	[33]

(l) Pharmacological activity: wound healing ability

Experiment design; (Experiment model; Assay; Test subject)	Extract	Plant part	Plant Source	Extract dose	Result	Reference
in vitro; Migration rate of HGF at 0 h, 2 h, 4 h, and 6 h after scratching; Human gingival fibroblast (HGF)	Chloroform, Hexane; Sun dry	Leaf	Indonesia	10, 25, 50 and 100 μg/mL	Culture supplemented with 10 μg/mL of chloroform extract give the fastest HGF migration rate and wound recovery	[11]

(m) Pharmacological activity: protective effect on plasmid DNA

Experiment design; (Experiment model; Assay; Test Subject)	Extract	Plant part	Plant Source	Extract dose	Result	Reference
in vitro; DNA integrity assay; Supercoiled plasmid DNA react with O2^•-generated via a riboflavin photoreaction treatment	70% Ethanol; Freeze dry; Sonication	Leaf	Perak; Malaysia	10000, 1000, 100, 10 and 0 μg/mL; Green tea	Extract reduced DNA cleavages, retained high levels of super-coiled plasmid DNA integrity and exhibited protection up to 50 min	[64]

(n) Pharmacological activity: lipid elevated inhibition activity

Experiment design; (Experiment model; Assay)	Extract	Plant part	Plant Source	Extract dose; Positive control	Result	Reference
in vitro; Porcine pancreatic lipase inhibition assay	Methanol; Freeze dry; Sonication	Leaf	Malaysia	100 μg/mL; Orlistat	Ineffective, −22.56% indicates a promotion of pancreatic lipase activity	[21]

(o) Pharmacological activity: oral mucositis and stomatitis treatment

Experiment design; (Experiment model; Infection; Assay; Test subject; Route of administration/Prescription)	Extract	Plant Source	Positive control	Result	Reference
Clinical; Radiation induced oral mucositis; Single-blinded-Randomized trial; 60 patients; Apply 2 drops drip into mouth or on the lesion 3 to 5 times daily from first to last day of radiation	Glycerin payayor	Thailand	Benzydamine hydrochloride; (Difflam, 3M, Australia)	The time onset of oral mucositis in the payayor group was significantly later, and its severity and pain score were less than those of the benzydamine group throughout the study period.	[10]
Clinical; Reccurent aphthous stomatitis; Double-blinded-controlled trial; 43 patients; apply to the ulcer 4 times daily	C. nutans in orabase	Thailand	Triamcinolone acetonide, placebo	C. nutans in orabase provide better healing of the ulcer as compared to placebo but efficacy was lesser compared to triamcinolone acetonide in orabase	[91]