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Journal of Drug Delivery and Therapeutics

Open Access to Pharmaceutical and Medical Research

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Open Access Full Text Article   Research Article

Xanthine Oxidase Inhibition and Anti-Lung Cancer Effects of the Lonicera japonica Thunb–Paris polyphylla var. chinensis Herbal Pair

Quang-Ung Le ¹, Thi-Lan Dinh ²*

¹ Thai Nguyen University of Agriculture and Forestry, Thai Nguyen, Vietnam

² Thai Nguyen University of Science, Thai Nguyen, Vietnam

 

 

INTRODUCTION

The advancement of adjunctive therapeutic approaches for the management of gout has crucial clinical significance. In the body, xanthine oxidase (XO) is a critical enzyme catalyzing the oxidation of xanthine to uric acid which may cause hyperuricemia^1,2. One of the basic therapeutic strategies in the limitation of uric acid overproduction in gout care and its related complications is that many recently synthesized antihyperuricemic drugs had been developed and invented. However, several clinically applied uric acid–lowering drugs elicited side effects and toxicity. Therefore, the study of useful natural products for the gout prevention and treatment have been highly required.Bottom of Form Lung cancer evolves into a dominant factor of cancer-related mortality worldwide³. The modern therapy methods encompassing surgical resection, chemotherapy, radiotherapy and targeted therapies. However, these modalities are often connected with stringent side effects and non-responsiveness⁴. The natural medicinal products are promising applicants with a safe and effective anticancer potential as well as sustainable health protection. These products have been proven capacities of modulating censorious oncogenic pathways, activating apoptosis, angiogenesis inhibition, and immune surveillance boostion in compared with standard chemotherapeutics^5,6.

In family Caprifoliaceae, Lonicera japonica Thunb. is a valuable herb, and its flowers (LTF) are utilized in many traditional medicine remedies. One of the a mainly active chemical compounds attracting appreciable attention is chlorogenic acid. Advanced pharmacological investigations had illustrated that L. japonica has been exhibited to have benefit biological activities, such as antibacterial, anti-inflammatory, antiviral, anti-endotoxin, lipid-lowering, antipyretic, and other merit therapeutic effects⁷. Paris polyphylla var. chinensis, a rarely perennial herb, belongs to family Trilliaceae. Its rhizomes (PPR) have long been applied to cure inflammation, sore throat, snake envenomation, severe pain, and convulsive syndromes⁸. Many recently pharmacological researches disclosed that phytochemical compounds isolated from the PPR and its crude extracts owned potential biological activities, for instance anticancer⁹, antioxidant¹⁰, antibacterial¹¹, hemostatic¹², and anthelmintic effects¹³. The PPR have been reported to strongly exhibit potential of cancer cell lines, for example CCRF-CEM leukemia cells, ECA109, CaEs-17, HL-60, HepG2, BGC-823, as well as LoVo, SW-116 and murine lung adenocarcinoma cells^{14,15,16,17,18}. In Vietnamese folk medicine, the LTF have been widely applied for gout cure. Especially, the LTF was combined with PPR for lung cancer cure with verified clinical cases. However, the XO inhibitory potential of chlorogenic acid in the LTF has not been reported. Moreover, the XO and A549 cell growth inhibitory capacity of combination of two herbs are currently deficient in investigations. Some works have illustrated that herbal extracts, particularly in polyherbal formulations, supply superior therapeutic effects when compared with equivalent doses of single herbs administered alone, underscoring the significance of synergistic interactions in herbal medicine^19-21. This work was conducted to evaluate the XO inhibitory and A549 cell growth proliferation activities of the herbal pair Paris polyphylla var. chinensis and Lonicera japonica Thunb., as well as their isolated chemical constituents. 

MATERIALS AND METHODS

Chemicals, reagents and instruments

The chemicals and reagents comprised xanthine oxidase (XO), xanthine (>99%), allopurinol, chlorogenic acid, and ferulic acid. Organic solvents were used for extraction and chromatographic separation such as ethanol 95% (EtOH), methanol 95% (MeOH), ethyl acetate (EtOAc), dimethyl sulfoxide (DMSO), and methylene chloride (DCM). Additional reagents are phosphate buffer, Tris–HCl, NaCl, Tween 20, Tris Buffered Saline with Tween 20 (TBST), nonfat dry milk, sodium dihydrogen phosphate, disodium hydrogen phosphate, hydrochloric acid, and sodium hydroxide. Reagents for cell-based assays are Kaighn’s modification of Ham’s F-12 medium (KMH F-12), fetal bovine serum (FBS), phosphate-buffered saline (PBS), and penicillin–streptomycin. The MTT reagent [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] and other reagents. The main instruments are such as rotary evaporator; normal-phase silica gel, reversed-phase silica gel (RP-18), and Sephadex LH-20 for column chromatography; TLC plates (silica gel 60 F254 and RP-18 F254s); and high-performance liquid chromatography (HPLC) systems, along with other equipments.Top of Form

Bottom of Form

Plant materials

The LTF and PPR of Paris polyphylla var. chinensis were gathered from Thai Nguyen and Lai Chau provinces, Vietnam. The plant materials were taxonomically authenticated by Dr. Quang Ung Le, in partnership with the Classification and Identification Committee of the Faculty of Agriculture Technology, Thai Nguyen University of Agriculture and Forestry. This committee has nine experts specializing in plant taxonomy, botany, pharmacognosy, and herbal sciences.

Extraction and isolation

The LTF in flowering stages (green bud stage-A, white bud and silver flowering stage-B and golden flowering stage-C) were washed by distilled water and dried in oven at 45 ^oC for 24 hours (hr). Five grams of 50-mesh sieved dry powder were separately extracted two times with EtOH at concentration of 50% (vol/vol) (E50A, E50B and E50C, respectively with flowering stages) at 30 ^oC by ultrasonic and then was filtered to give filtrate combinations and concentrated in a rotary evaporator at 45 ^oC. The dehydrated fractionation yield was calculated by using electronic scale, then dissolved in dimethyl sulforide (DMSO) to give a regular concentration to perform continue experiments

Two kilograms of PPR powder were exhaustively extracted with 15 L 95% MeOH by ultrasonic (three times, 90 min each at 40 °C). The combined MeOH extracts were condensed under reduced pressure to obtain a dark crude extract (PPR-MeOH extract 362.0 g). Then, this extract was suspended in distilled water (1 L) and successively partitioned with EtOAc to yield an EtOAc fraction (EtOAc, 82.6 g). The EtOAc extract was subjected to silica gel column chromatography using a gradient elution system of dichloromethane/methanol/ water (from 100% DCM to MeOH/H₂O, 1/2/0.01, v/v) to give six fractions (PR-1→PR-6). Fraction PR-1 (0.6 g) was further purified by silica gel column chromatography eluted with DCM/MeOH (from 100% DCM to MeOH 10:1, v/v), gaining compound PR-1A, identified as β-sitosterol-3-O-β-D-glucopyranoside (44.3 mg). Fraction PR-4 (13.4 g) was separated by reversed-phase C18 column chromatography using n-hexane/ethyl acetate (5:1, v/v) as the eluent to obtain compound PR-4D (50.0 mg) chromatographed on a silicagel column to give compound PR-4D1 (quercetin, 22.0 mg). The separation procedures of compounds PR-1A and PR-4D1 are briefed in Figure 1

 

 

Figure 1. Schematic diagram of PR-1A and PR-4D1 isolation from ethyl acetate fraction

 

 

High performance liquid chromatography analysis

The phenolic constituents in the E50A, E50B and E50C extracts were quantitatively analyzed by HPLC. The system (Hitachi, Tokyo, Japan) was installed with a Chromaster 5110 pump, Chromaster 5210 autosampler, Chromaster 5430 diode array detector, and a Mightysil RP-18 GP column (4.6 × 250 mm; Kanto Chemical Co. Inc., Tokyo, Japan). The mobile phase comprised solvent A (distilled water) and solvent B (MeOH), both adjusted to pH 2.8 with H₃PO₄. The following gradient program was performed: 0-3 min, 5% B; 3-6 min, 7-10% B; 6-10 min, 10-15% B; 10-20 min, 15-20% B; 20-25 min, 20-25% B; 25-30 min, 25-28% B; 30-35 min, 28-30% B; 35-40 min, 30-40% B; 40-45 min, 40-42% B; 45-50 min, 42-45% B; 50-60 min, 45-30% A; and 60-65 min, 35-30% B. Chromatographic settings were performed as follows: flow rate of 1.0 mL/min, injection volume of 20 µL, and UV detection at 280 nm. The column temperature was continued at 40°C during the analysis. The phenolic constituents were resulted in by interpolation using the linear regression plot from the standard component solution. 

Xanthine oxidase inhibition assay

The XO inhibitory experiments of compounds and extracts were performed according to Chen et al. (2010)²². Various concentrations of samples were prepared in 1% DMSO. The assay mixture comprised 40 μL of 1% DMSO (blank) or compound solution and 60 μL of XO enzyme solution (0.02 U/mL in 50 mM PBS, pH 7.5), freshly prepared before the experiment. After incubation at 37°C, the absorbance was determined at 295 nm after 45 min. The positive control compound is allopurinol. The XO inhibitory ability was quantified: inhibition (%) = (A₀ − Aₜ)/A₀ × 100%, in which A₀ and Aₜ correspond to the absorbance of the blank and the test sample, respectively. The IC₅₀ value was calculated as the dosage required to inhibit XO by 50%.

Cytotoxic effects of compounds and fraction mixtures

Cell culture

A549 cells were cultured in KMH F-12 medium added with 10% FBS, 100 U/mL penicillin, and 100 µg/mL streptomycin. The culture medium was newed every 72 hr, and the cells were stably incubated in a 5% CO₂ and 37 °C condition incubator.

Cell Viability Assay                                                                                                                                    

A549 cells were cultured in 96-well plates with a 5 × 10³ density per well in 200 µL of culture medium, with each assay performed in three times. Following a 24 hr attachment period, the cells were exposed to compounds or extracts at the prepared dosages in serum-free medium for 48 hr. The positive control compound in this assay is fluorouracil, while cells treated with 0.2% DMSO functioned as the vehicle control. Cell viability was subsequently evaluated using the MTT assay according to an earlier reported method²³.

Western blot analysis

The PPR-MeOH and E50B mixture (at ratio 1:1), presenting the strongest anticancer potential (Table 3), was used to induce apoptosis in A549 cells. Whole cell protein extracts were prepared using 1× RIPA buffer supplemented with a complete protease inhibitor cocktail. The centrifugation (12,000 × g for 20 minutes at 4 °C) was used to purify lysates to give the supernatants. Protein concentrations were quantified by the Bradford assay, the proteins mixing with 2×sample loading buffer were incubated at 95 °C for 5 min, and separated on 12% polyacrylamide gels using the Mini-Protean 3 electrophoresis system (Bio-Rad). Proteins were subsequently transferred onto nitrocellulose membranes which were then blocked with 5% non-fat milk/TBST (10 mM tris-HCl, pH 7.5, 150 mM NaCl, 0.1% tween 20) for 2 hr at room temperature and then cultured overnight at 4 °C with the relevant primary antibodies. Following washing with blocking buffer three times for 0.5 hr, the membranes were incubated for 2 hours with horseradish peroxidase–conjugated goat anti-mouse or anti-rabbit IgG secondary antibodies. After extensive washing thrice for 1 hr with Tris-buffered saline Tween 20, immunoreactive bands were detected by Enhanced chemiluminescence Detection Kit using a LAS-3000 luminescent imaging system. Antibody dilutions for Western blot analysis were as follows: β-actin (1:3000); caspase-3, caspase-9, and p53 (1:500), as formerly described²⁴.

Statistical analysis

The experimental data were analyzed using one-way analysis of variance (ANOVA) to evaluate statistically significant differences among treatment groups. Post-hoc comparisons were carried out using the least significant difference (LSD) test. All statistical procedures were performed with the SAS software package (SAS Institute, 1990), and significance was considered at p < 0.05.

RESULTS AND DISCUSSION

The results of the in vitro xanthine oxidase inhibitory activity test of PPR extracts showed that the EtOAc fraction exhibited the higher xanthine oxidase inhibitory activity compared to the H₂O fraction with an IC50 value of 38.15 µg/ml and 40.18 µg/ml, respectively.   Based on the in vitro xanthine oxidase inhibitory activity test, the ethyl acetate fraction was selected for analysis and chemical structure determination.

Physical constants and spectral data and structure elucidation of isolated compounds of PPR 

Compound 1 (PR-1A) was isolated as an amorphous, colorless solid, with a melting point of 269-270 °C. ¹H-NMR spectrum of PR-1A was displayed in figure 2a. In the ¹H-NMR spectrum of PR-1A, the characteristic signals of a sterol is identified, with 6 methyl groups at δH 0.72 (3H, s, H-18), 1.03 (3H, s, H-19), 0.94 (3H, d, J = 6.5 Hz, H-21), 0.79 (3H, d, J = 7.0 Hz, H-26), 0.87 (3H, d, J = 7.0 Hz, H-27) and 0.80 (3H, t, J = 7.0 Hz, H-29); one oxymethine group at δH 3.60 (1H, m, H-3); a trisubstituted double bond signal at δH 5.24 (1H, brs, H-6) and  two protons of an exocyclic double bond at δH 5.17 (1H, dd, J =15; 8.5 Hz, H- 22) and δH 5.04 (1H, dd, J =15.0; 8.5 Hz, H-23). The ¹³C-NMR appeared to have signals of 35 carbons with characteristic of a sterol skeleton: 6 methyl groups at δC 11.8 (C-18), 19.5 (C-19), 20.9 (C-21), 18.7 (C-26), 20.8 (C-27) và 11.9 (C-29); one oxymethine group at δC 75.7 (C-3); a trisubstituted double bond at δC 140.2 (C-5) 121.9 (C-6); and two methylene groups (CH₂) at δ 33.8 (C-22) and 25.9 (C-23). Present data suggested that PR-1A is β-sitosterol conjugated with one molecule of glucose.  Furthermore, an anomeric carbon signal was observed at δC 100.9 (C-1ʹ). The 1H-NMR spectrum also showed three oxymethine proton signals at δH 3.41 (1H, dd, J1=5.0; 8.5 Hz, H-2ʹ), 3.22 (1H, dd, J1=J2= 8.5, H-3ʹ), 3.41 (1H, dd, J1=J2= 8.5 Hz, H-4ʹ). Comparison with published spectral data allowed for the identification of the sugar moiety as β-D-glucopyranose. The spectral data of PR-1A were in full agreement with the evidenced spectral data for β-sitosterol-3-O-β-D-glucopyranoside²⁵. Chemical structure of β-sterol-3-O-β-D-glucopyranoside was shown in figure 3a.

Compound 2 (PR-4D1) was isolated as yellow crystalline solids having melting point at 313-314 °C. ¹H-NMR spectrum of PR-4D1 was shown in figure 2b. The ¹H NMR spectrum has characteristic features of a flavonol skeleton. In five aromatic ring proton signals featured for ABX- type, there are two meta-coupled proton signals at δ_H 6.51 (1H, d, J = 2.0 Hz) and 6.26 (1H, d, J = 2.0 Hz) assigned to H-8 and H-6, respectively, corresponding to the 5,7-dihydroxy structure of ring A; and three aromatic protons at δ_H 7.82 (1H, d, J = 2.0 Hz, H-2′), 6.99 (1H, d, J = 8.5 Hz, H-5′), and 7.69 (1H, dd, J = 8.5, 2.0 Hz, H-6′) belongs to ring B, which suggests substitution at the C-1′, C-3′, and C-4′ positions. The absence of a singlet corresponding to H-3 of the aromatic nucleus further supported substitution at the C-3 position. In the ¹³C-NMR spectrum, there is a signal at δ_C 136.7, which is feature of a flavonol bearing a OH group at C-3. The ¹³C-NMR indicated that PR-4D1 comprises 15 carbon atoms. Of these, five CH groups featured aromatic carbons at δ_C 94.2 (C-6), 99.1 (C-8), 115.7 (C-2′), 116.2 (C-5′), and 121.4 (C-6′) ppm and ten quaternary carbons in which the signal at δ_C 176.5 ppm featured for a conjugated carbonyl group (C=O) in a flavonoid skeleton. Four quaternary carbon signals at δ_C 145.8, 146.9, 162.3, and 164.9 ppm were assigned to aromatic carbons bonded to OH groups, corresponding to C-3′, C-2, C-5, and C-7, respectively. These spectroscopic data clearly support that PR-4D1 is a flavonoid. On the basis of the comparison with the published studies allowed to conclude that the identified PR-4D1 as quercetin²⁶. Chemical structure of quercetin was shown in figure 3b. 

 

Figure 2. ¹H-NMR spectra of β-sitosterol-3-O-β-D-glucopyranosid and quercetin

 

 

 

Figure 3. Chemical structure of β-sterol-3-O-β-D-glucopyranoside (a), quercetin (b).

 

 

The chlorogenic acid identification of LTF in flowering stages

 

Figure 4. HPLC chromatograms of chlorogenic acid and related phenolic compounds. (a) Standard phenolic compounds; (b) green bud stage; (c) white bud and silver flowering stages; (d) golden flowering stage.

Abbreviations: 4-HB, 4-hydroxybenzoic acid; Chlo, chlorogenic acid; Benzo, benzoic acid; p-Cua, p-coumaric acid; Cafe, caffeic acid; Feru, ferulic acid; Rosma, rosmarinic acid.

 

The chromatographic profiles of the LTF at different flowering stages are displayed in Figure 4b-d. Chlorogenic acid and ferulic acid were quantified in LTF, with chlorogenic acid being the main active constituent. The content of chlorogenic acid varied across the flowering stages, following the order: white bud and silver flowers (268.43 ± 0.02 mg/g dry extract) > green flowers (202.4 ± 0.01 µg/g dry extract) > yellow flowers (104.3 ± 0.02 µg/g dry extract) (Figure 5), and the content of ferulic acid also is highest in white bud and silver flowering stage (167.23±0.04 µg/g dry extract). More than 140 compounds have been isolated and identified from L. japonica, with major constituents such as chlorogenic acid, isochlorogenic acid, caffeic acid, hexadecanoic acid, and myristic acid⁷. Particularly, ferulic acid was determined for the first time in LTF. 

Figure 5. Chlorogenic acid content at different flowering stages of Lonicera japonica Thunb. Values are expressed as mean ± SD (n = 3). Different superscript letters indicate statistically significant differences (p < 0.05).

Xanthine oxidase enzyme inhibiting capacity of compounds and extract mixtures

Table 1.  Xanthine oxidase enzyme inhibiting capacity of compounds and mixtures

Note. Values are expressed as mean ± SD (n = 3). Different superscript letters indicate statistically significant differences (p < 0.05). ^*Positive control substance.

The XO inhibiting capacity in vitro of compounds and extracts was evaluated by the IC₅₀ values (Table 1). The results were found that XO inhibitory effect reduced in the following order: ferulic acid > chlorogenic acid > quercetin> E50B> β-sitosterol-3-O-β-D-glucopyranoside > PPR-MeOH. As of now, β-sitosterol-3-O-β-D-glucopyranoside, PPR-MeOH and E50B were firstly tested XO inhibition capacity.

Extract mixture induced growth inhibition in A549 cells

PPR-MeOH, E50B, and their combined formulation (PPR-MeOH and E50B, 1:1, v/v) significantly induced cytotoxic effects against A549 cells. The growth-inhibitory potency against A549 cells reduced in the following order: fluorouracil > PPR-MeOH: E50B mixture (1:1) > PPR-MeOH > E50B (Table 2). Especially, the combined extracts presented the stronger antiproliferative effect compared to either extract alone, which recommended a potential synergistic or additive interaction between PPR-MeOH and E50B. In former works, Lonicera japonica Thunb had been evidenced significantly enhancing bioactivity when combined with other medicinal herbs such as co-administration with Oroxylum indicum²⁷, in combination with Astragalus membranaceus²⁸, and Magnolia obovata²⁹. These synergistic effects should furnish insightful understanding support for the rational development of multi-herb formulations for natural remedy applications. The previously in vitro researches strongly evidenced that both crude extracts and isolated compounds from P. polyphylla var. chinensis displayed tumor growth–suppressive activity of cell lines such as MCF-7 breast cancer cell^30,31,32, HepG2³¹, MDA-MB-231³², HL-60, A549/ATCC, SW-620, melanoma M14, and renal cancer 786-0 cell lines³³; However, further mechanistic researches and clinical validation are essential to fully establish its therapeutic potential.

Table 2. Cytotoxicity of extracts against A549 cells after 24 hr

Compounds	IC50 (µg/mL)
Flourouracil*	2.29 ± 0.22d
PPR-MeOH	6.32±0.17b
E50B	10.30±0.23a
PPR-MeOH and E50B miture (at ratio 1:1)	4.11±0.16c

Note. Values are expressed as mean ± SD (n = 3). Different superscript letters indicate statistically significant differences (p < 0.05). ^*Positive control substance.

PPR-MeOH and E50B mixture altered the expression of apoptosis-related proteins A549 cells

The expression of apoptosis-related proteins in A549 cells was detected by Western blotting (Figure 6). The results highlighted that treatment with the PPR-MeOH and E50B mixture markedly activated caspase-3 and caspase-9 and enhanced cleavage of p53. The tumor suppressor gene p53 is a vital regulator of apoptosis and a crucial barrier against tumor development^34,35. Activated caspase-9 and caspase-3 are important executors of the intrinsic apoptotic pathway in cancer cells. Caspase-9 is activated following mitochondrial Cytochrome c release, subsequently triggering caspase-3 activation, which mediates proteolytic cleavage of critical cellular substrates and results in apoptotic cell death^36,37. These discoveries strongly emphasize that the combination of LTF and PPR induces apoptosis in A549 cells, which provides robust evidence for the lung anticancer potential of the LTF–PPR mixture and recommends a scientific rationale for the traditional use of them to treat lung cancer in Vietnamese folk medicine.

Figure 6. PPR-MeOH and the E50B mixture significantly modulated the expression of apoptosis-related proteins, including caspase-3, caspase-9, and p53, in A549 cells after 24 h of treatment

CONCLUSION

In summary, ferulic acid is the first to identify from the LTF. The XO inhibitory capacity of the isolated compounds and extracts, and A549 cell growth-inhibitory effects of the LTF–PPR mixture are also firstly investigated. PPR-MeOH and E50B miture (at ratio 1:1) presented the stronger antiproliferative effect compared to either extract alone. Authors call for research strategy on the synergistic effects by demonstrating molecular mechanisms models and perform rigorous quantitative evaluation of the synergistic effects of the herbal pair.

Acknowledgment: The authors would like to express their sincere gratitude to the Classification and Identification Committee of the Faculty of Agriculture Technology, Thai Nguyen University of Agriculture and Forestry, for their valuable support.

Conflict of interest: There are no competing financial interests.

REFERENCES

1.  Le, UQ. The review on medicinal herbs in the treatment of gout through xanthine oxidase inhibitory activity: Call for more research strategy in the future. Journal of Applied Pharmaceutical Science. 2024; 14(4): 001-013. https://doi.org/10.7324/JAPS.2024.16031

2.  Wang H, Zhang H, Zhang X, Yin Y, Ding G, Tang X, et al. Identification of coniferyl ferulate as the bioactive compound behindthe xanthine oxidase inhibitory activity of chuanxiong rhizome. Journal of Functional Foods, 2023;100:105378. https://doi.org/10.1016/j.jff.2022.105378

3.  Bray F, Ferlay J, Soerjomataram I, Siegel RL, Torre LA, Jemal A. Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA: A Cancer Journal for Clinicians, 2018; 68(6): 394-424. https://doi.org/10.3322/caac.21492 PMid:30207593

4.  Sung H, Ferlay J, Siegel RL, Laversanne M, Soerjomataram I, Jemal A, Bray F. Global cancer statistics 2022: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA: A Cancer Journal for Clinicians, 2023; 73(3), 209-249. https://doi.org/10.3322/caac.21660 PMid:33538338

5.  Cragg GM, Newman DJ. Natural products: A continuing source of novel drug leads. Biochimica et Biophysica Acta (BBA) - General Subjects, 2013; 1830(6): 3670-3695. https://doi.org/10.1016/j.bbagen.2013.02.008 PMid:23428572 PMCid:PMC3672862

6.  Newman DJ, Cragg GM. Natural products as sources of new drugs over the nearly four decades from 1981 to 2019. Journal of Natural Products, 2020; 83(3): 770-803. https://doi.org/10.1021/acs.jnatprod.9b01285 PMid:32162523

7.  Shang X, Pan H, Li M, Miao X, Ding H. Lonicera japonica Thunb.: ethnopharmacology, phytochemistry and pharmacology of an important traditional Chinese medicine. Journal of Ethnopharmacology, 2011; 138(1): 1-21. https://doi.org/10.1016/j.jep.2011.08.016 PMid:21864666 PMCid:PMC7127058

8.  Ding YG, Zhao YL, Zhang J, Zuo ZT, Zhang QZ, Wang YZ. The traditional uses, phytochemistry, and pharmacological properties of Paris L. (Liliaceae): A review. Journal of Ethnopharmacology, 2021; 278:114293. https://doi.org/10.1016/j.jep.2021.114293 PMid:34102270

9.  Jing S, Wang Y, Li X, Man S, Gao W. Chemical constituents and antitumor activity from Paris polyphylla Smith var. yunnanensis. Natural Product Research, 2016; 31:660-666. https://doi.org/10.1080/14786419.2016.1219861 PMid:27687140

10.  Shian S, Zhou X, Shiling F, Handong W, Jing L, Lijun Z, Yuan M, Huang Y. Ding C. Structural elucidation and antiaging activity of polysaccharide from Paris polyphylla leaves. International Journal of Biological Macromolecules, 2018;107:1613-1619. https://doi.org/10.1016/j.ijbiomac.2017.10.026 PMid:28993294

11.  Qin XJ, Sun DJ, Ni W, Chen CX, Hua Y, He L, Liu HY. Steroidal saponins with antimicrobial activity from stems and leaves of Paris polyphylla var. yunnanensis. Steroids, 2012; 77: 1242-1248. https://doi.org/10.1016/j.steroids.2012.07.007 PMid:22846376

12.  Wei JC, Gao WY, Yan XD, Wang Y, Jing SS, Xiao PG. ChemInform Abstract: Chemical Constituents of Plants from the Genus Paris. Chemistry & Biodiversity, 2014; 45:1277-1297. https://doi.org/10.1002/cbdv.201300083 PMid:25238072

13.  Wang GX, Han J, Zhao LW, Jiang DX, Liu YT, Liu XL. Anthelmintic activity of steroidal saponins from Paris polyphylla. Phytomedicine, 2010; 17:1102-1105. https://doi.org/10.1016/j.phymed.2010.04.012 PMid:20576414

14.  Kang LP, Liu YX, Eichhorn T, Dapat E, Yu HS, Zhao Y, Xiong CQ, Liu C, Efferth T, Ma BP. Polyhydroxylated steroidal glycosides from Paris polyphylla. Journal of Natural Products, 2012; 75:1201-1205. https://doi.org/10.1021/np300045g PMid:22663190

15.  Li FR, Jiao P, Yao ST, Sang H, Qin SC, Zhang W, Zhang YB, Gao LL. Paris polyphylla Smith extract induces apoptosis and activates cancer suppressor gene connexin26 expression. Asian Pacific Journal of Cancer Prevention, 2012; 13:205-209. https://doi.org/10.7314/APJCP.2012.13.1.205 PMid:22502669

16.  Zhao Y, Kang LP, Liu YX, Liang YG, Tan DW, Yu ZY, Cong YW, Ma BP. Steroidal saponins from the rhizome of Paris polyphylla and their cytotoxic activities. Planta Medica, 2009; 75: 356-363. https://doi.org/10.1055/s-0028-1088380 PMid:19085682

17.  Sun J, Liu BR, Hu WJ, Yu LX, Qian XP. In vitro anticancer activity of aqueous extracts and ethanol extracts of fifteen traditional Chinese medicines on human digestive tumor cell lines. Phytotherapy Research: An International Journal Devoted to Pharmacological and Toxicological Evaluation of Natural Product Derivatives, 2007; 21:1102-1104. https://doi.org/10.1002/ptr.2196 PMid:17639550

18.  Man SL, Gao WY, Zhang YJ, Ma CY, Yang L, Li YW. Paridis saponins inhibiting carcinoma growth and metastasis in vitro and in vivo. Archives of Pharmacal Research, 2011; 34: 43-50. https://doi.org/10.1007/s12272-011-0105-4 PMid:21468914 PMCid:PMC10950667

19.  Leonard SS, Cutler D, Ding M, Vallyathan V, Castranova V, and Shi XL. Antioxidant properties of fruit and vegetable juices: more to the story than ascorbic acid. Annals of Clinical & Laboratory Science, 2002; 32: 193-200.

20.  Scholey AB, and Kennedy DO. Acute, dose-dependent cognitive effects of Ginkgo biloba, Panax ginseng and their combination in healthy young volunteers: differential interactions with cognitive demand. Human Psychopharmacology: Clinical and Experimental, 2002; 17: 35-44. https://doi.org/10.1002/hup.352 PMid:12404705

21.  Zhang AH, Sun H, and Wang XJ. Potentiating therapeutic effects by enhancing synergism based on active constituents from traditional medicine. Phytotherapy Research, 2014; 28: 526-533. https://doi.org/10.1002/ptr.5032 PMid:23913598

22.  Chen CH, Chen PY, Wang KC, Lee CK. Rapid identification of the antioxidant constituent of Koelreuteria henryi. Journal of the Chinese Chemical Society, 2010; 57: 404-410. https://doi.org/10.1002/jccs.201000060

23.  Le QU, Horng LL, Ming CW. The isolation, structural characterization, and anticancer activity from the aerial parts of Cymbopogon flexuosus. Journal of food biochemistry, 2019;43(2): e12718. https://doi.org/10.1111/jfbc.12718 PMid:31353668

24.  Nho KJ, Chun JM, Kim HK. Agrimonia plilosa ethanol extract induces apoptotic cell death in HepG2 cells. Journal of Ethnopharmacology, 2011; 138: 358-363. https://doi.org/10.1016/j.jep.2011.09.008 PMid:21945237

25.  Peshin T, Kar HK. Isolation and Characterization of β-Sitosterol-3-O-β-D-glucoside from the Extract of the Flowers of Viola odorata. Journal of Pharmaceutical Research International, 2017; 16(4):1-8. https://doi.org/10.9734/BJPR/2017/33160

26.  Awasthi, A. Isolation and Characterization of Quercetin from Bambusa arundinacea. Journal of Pharmaceutical Research International, 2021; 33(54B): 256-262. https://doi.org/10.9734/jpri/2021/v33i54B33784

27.  Le TDH, Phuong NTM, On TM. The anti-inflammatory effect of honeysuckle flowers (Lonicera japonica) in combination with Oroxylum indicum. VNU Journal of Science: Medical and Pharmaceutical Sciences, 2020; 36(2): 25-32. https://doi.org/10.25073/2588-1132/vnumps.4222

28.  Yeh CF, et al. Honeysuckle and Astragalus membranaceus suppress SARS-CoV-2 entry and COVID-19-related cytokine storm in vitro. Journal of Ethnopharmacology, 2022; 289: 115000. https://doi.org/10.3389/fphar.2021.765553 PMid:35401158 PMCid:PMC8990830

29.  Kim JH, Lee SY, Park JM. Synergistic antimicrobial effect of Lonicera japonica and Magnolia obovata extracts and their potential as plant-derived natural preservatives. Food Control, 2018; 90: 292-300.

30.  Nguyen TD, Vinh LB, Phong NV, et al. Steroid glycosides isolated from Paris polyphylla var. chinensis aerial parts and paris saponin II induces G1/S-phase MCF-7 cell cycle arrest. Carbohydrate Research, 2022; 519:108613. https://doi.org/10.1016/j.carres.2022.108613 PMid:35752103

31.  Hu X, Zhang L, Wang T, et al. Study of chemical compositions and anticancer effects of Paris polyphylla var. chinensis leaves. Molecules, 2022; 27(9):2724. https://doi.org/10.3390/molecules27092724 PMid:35566077 PMCid:PMC9100081

32.  Hoang TPT, Ly HT, Nguyen TT, Do TH, Le VM. Standardized extracts of Paris polyphylla var. chinensis rhizomes suppress breast cancer cell migration. Journal of Medicinal Materials, 2025; 30(5):375-384. https://doi.org/10.63240/jmm-nimm.2025.5.173

33.  Mimaki Y, Kuroda M, Obata Y, Sashida Y, Kitahara M, Yasuda A, Lao AN. Steroidal saponins from the rhizomes of Paris polyphylla var. chinensis and their cytotoxic activity on HL-60 cells. Natural Product Letters, 2000; 14(5): 357-364. https://doi.org/10.1080/10575630008043768

34.  Levine AJ. p53, the cellular gatekeeper for growth and division. Cell, 1997; 88(3), 323-331. https://doi.org/10.1016/S0092-8674(00)81871-1 PMid:9039259

35.  Chipuk JE, Maurer U, Green DR, Schuler M. Mechanisms regulating mitochondrial outer membrane permeabilization. Oncogene, 2004; 23(16): 2850-2860. https://doi.org/10.1038/sj.onc.1207534 PMid:15077148

36.  Li P, Nijhawan D, Budihardjo I, Srinivasula SM, Ahmad M, Alnemri ES, Wang X. (1997). Cytochrome c and dATP-dependent formation of Apaf-1/caspase-9 complex initiates an apoptotic protease cascade. cell, 1997; 91(4): 479-489. https://doi.org/10.1016/S0092-8674(00)80434-1 PMid:9390557

37.  Porter AG, Jänicke RU. Emerging roles of caspase-3 in apoptosis. Cell Death and Differentiation, 1999; 6(2): 99-104. https://doi.org/10.1038/sj.cdd.4400476 PMid:10200555