Phytochemical standardization of panchavalkala: An ayurvedic formulation and evaluation of its anticancer activity in cervical cancer cell lines
Shama Aphale1, Savita Pandita1, Prerna Raina1, JN Mishra2, Ruchika Kaul-Ghanekar1
1 Cancer Research Lab, Interactive Research School for Health Affairs, Bharati Vidyapeeth (Deemed to be University), Pune, Maharashtra, India
2 Bharat Sewa Sansthan, Moti Mahal, Rana Pratap Marg, Lucknow, Uttar Pradesh, India
|Date of Submission||11-May-2018|
|Date of Acceptance||28-Jun-2018|
|Date of Web Publication||21-Nov-2018|
Interactive Research School for Health Affairs, Bharati Vidyapeeth (Deemed to be University), Katraj-Dhankawadi, Pune-Satara Road, Katraj, Pune - 411 043, Maharash
Source of Support: None, Conflict of Interest: None
| Abstract|| |
Background: Cervical cancer is the most common malignant disease affecting women worldwide. The currently available therapies for cancer, even though effective, affect the patient's health severely due to the associated side effects. Thus, nowadays, complementary/alternative medicines are being extensively researched upon for their use as an adjunct therapy. Panchavalkala, an Ayurvedic formulation, is traditionally being used as a douche in leukorrhea and other gynecological diseases. Objective: The objective of the study was to phytochemically standardize aqueous extract of Panchavalkala (PVaq) and evaluate its anticancer activity against human cervical cancer cell lines. Materials and Methods: The phytochemical characterization of PVaq was done by liquid chromatography–mass spectrometry (LCMS) technique. The effect of PVaq on the viability of SiHa and HeLa cells was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium Bromide dye assay. The effect of the extract on growth kinetics was evaluated by trypan blue dye exclusion method and soft agar assay. Results: LCMS analysis showed presence of 77 compounds, of which 15 major compounds included proanthocyanidin B1, chlorogenic acid, caffeic acid, epicatechin, leucopelargonidin 3-O-alpha-L-rhamno-beta-D-glucopyranoside, leucocyanidin, naringenin-7-o-glucoside, mesoinositol, catechin, vogelin E, mesoinositol, behenic acid, bergenin, acacetin, and gallic acid. PVaq significantly (P < 0.001) reduced the viability of SiHa and HeLa cells with an IC50 of 125.8 and 96.0 μg/ml, respectively. It also reduced the growth of cervical cancer cells in a dose- and time-dependent manner. Conclusion: This preliminary data suggests that PVaq exhibits potential anticancer activity and warrants further studies for detailed elucidation of its mechanism of action.
Abbreviations used: PVaq: Aqueous extract of Panchavalkala; LCMS: Liquid chromatography–mass spectrometry; HPV: Human papillomavirus; MTT: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide.
Keywords: Aqueous extract of Panchavalkala, cell viability, cervical cancer, growth kinetics, liquid chromatography–mass spectrometry
|How to cite this article:|
Aphale S, Pandita S, Raina P, Mishra J N, Kaul-Ghanekar R. Phytochemical standardization of panchavalkala: An ayurvedic formulation and evaluation of its anticancer activity in cervical cancer cell lines. Phcog Mag 2018;14:554-60
|How to cite this URL:|
Aphale S, Pandita S, Raina P, Mishra J N, Kaul-Ghanekar R. Phytochemical standardization of panchavalkala: An ayurvedic formulation and evaluation of its anticancer activity in cervical cancer cell lines. Phcog Mag [serial online] 2018 [cited 2021 Sep 26];14:554-60. Available from: http://www.phcog.com/text.asp?2018/14/58/554/245847
- In the present study, we have done phytochemical standardization of aqueous extract of Panchavalkala (PVaq) and evaluated its anticancer activity against cervical cancer cell lines. PVaq showed presence of the phytocompounds having reported antioxidant, anti-inflammatory, and anticancer activities
- PVaq decreased the viability of human papillomavirus-positive cervical cancer cells in a dose-dependent manner
- It altered the growth kinetics of the cancer cells in a dose- and time-dependent manner.
| Introduction|| |
Cervical cancer is the second most leading cause of cancer death among Indian women. Every year in India, 122,844 women are diagnosed with cervical cancer and 67,477 die from the disease. The persistent infection with human papillomavirus (HPV), notably type 16 and 18 has been found to be the primary cause for cervical cancer. The available conventional therapies which include surgery/chemotherapy/radiotherapy have greatly reduced the mortality; however, they are associated with serious adverse events., Nowadays, cancer research has shifted its focus towards identification of herbal based drugs that have no or minimum side effects; exhibit anticancer activity; and could be used as adjunct to conventional therapies.
Panchavalkala is an Ayurvedic formulation comprising equal proportions of bark materials of Ficus benghalensis, Ficus virens, Ficus religiosa, Ficus glomerata, and Thespesia populnea. Traditionally, it has been used for the treatment of female infertility and endometriosis-related problems.,,,,, The decoction has been extensively used as a douche in leukorrhea and other vaginal diseases.,,,, It has also been reported that regular douching of the genital tract with the decoction of Panchavalkala helps in reduction of symptoms related to vaginitis.,,,, In addition, the individual components of the formulation have well reported anticancer activity.,, In the present study, we have for the first time performed liquid chromatography–mass spectrometry (LCMS) profiling of aqueous extract of Panchavalkala (PVaq) and evaluated its anticancer activity against HPV16+ (SiHa) and HPV18+ (HeLa) cervical cancer cell lines. LCMS profiling revealed the presence of compounds with reported anti-inflammatory, antioxidant, and anticancer properties. Treatment of SiHa and HeLa cells with PVaq significantly reduced their viability and growth kinetics, thereby signifying the anticancer potential of the formulation.
| Materials and Methods|| |
Dulbecco's Modified Eagle's Medium (DMEM) powder, penicillin and streptomycin were purchased from Invitrogen/Gibco (Grand Island, NY, USA). Agarose was purchased from Gibco (DNA grade, BRL, CA, USA). Fetal bovine serum (FBS) and (3-4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Tissue culture plasticware was purchased from BD Biosciences (San Diego, CA, USA). The human cervical carcinoma cell lines, SiHa (HPV-16) and HeLa (HPV-18), were obtained from National Center for Cell Science, Pune, Maharashtra, India. The cells were grown in DMEM supplemented with 10% FBS, 2 mM L-glutamine, and antibiotics (100 units/ml penicillin and streptomycin). The cells were maintained in a humidified 5% CO2 incubator at 37°C.
Plant material and extraction
Barks of F. religiosa L. (Auth. 17-256), F. virens (Auth. 17-252), F. glomerata (Auth. 17-251), F. benghalensis (Auth. 17-250), and T. populnea (Auth. 17-253) were collected from Pune, Maharashtra, India. Botanical identification of plant material was carried out at Agharkar Research Institute, Pune, Maharashtra, India. The barks of all the five plants were chopped into small pieces, shade dried at ambient temperature, and stored in airtight container. It was ground into coarse powder in a grinder whenever required. The powders from each bark were taken in the 1:1 ratio for extract preparation. The extract obtained was centrifuged at 13,000 rpm for 15 min, and the supernatant was filtered through Swiney filter (pore size, 0.45 μm) and stored at −80°C until further use. Aqueous extract of the mixture of powders was prepared as per the standard procedure for preparation of kwath, i.e., aqueous extract given in Ayurvedic Formulary of India (deduced from Charaka Samhita, Sutrasthan Adhyay 4;8).
Liquid Chromatography–mass spectroscopy
Non-targeted and targeted HPLC-MS QTOF analysis was performed as described earlier. Briefly, 10 μL of sample was injected onto an Agilent 1290 HPLC system having Zorbax Eclipse Plus C18 column (2.1 mm × 100 mm, 1.8 μm particle sizes). The mobile phases consisted of (A) water and (B) acetonitrile (LCMS grade, J. T. Baker) with flow rate of 0.3 mL/min and 95:5 acetonitrile/water at a flow rate of 0.7 mL/min. Both mobile phases were modified with 0.1% (v/v) formic acid for MS analysis in positive mode and with 5 mm ammonium acetate for analysis in negative mode. The chromatographic conditions utilized for the study consisted of the first 5 min run isocratically at 5% B; a gradient of B from 95% to 5% was applied from 5 min to 30 min, followed by 3 min isocratically at 100%. MS analysis was performed on an Agilent 6530 Quadrupole time-of-flight spectrometer fitted with an electrospray ionization source in both positive and negative mode. Data were analyzed using Mass Hunter Qualitative Analysis Software Package (Agilent Technologies) and online database Metlin. Blanks using each of the solvent extraction systems were analyzed using “Find by Molecular Feature” algorithm in the software package to generate a compound list of molecules with significant abundances >10,000 counts. This was then used as an exclusion list to eliminate background contaminant compounds from the analysis of the extracts. The data were analyzed using “Find by Molecular Feature” function to generate a list of compounds with empirical formula in the extracts. Compound lists were then screened against online mass databases; METLIN Metabolomics Database and MassBank Database.
Cell viability assay
SiHa and HeLa cells were seeded at a density of 1 × 105 cells/ml in 96-well plates. The cells were treated with different concentrations (0, 10, 20, 40, 80, 160, 320, and 640 μg/ml) of PVaq in each well in triplicates for 24 h. Cell viability was determined by MTT assay as described previously.,
Cell growth analysis
SiHa and HeLa cells were seeded at a density of 1 × 105 cells/ml in 24-well plates in triplicates. Next day, the cells were treated with different concentrations of PVaq (0–320 μg/ml) for 24, 48, and 72 h. The cells were harvested and counted for viability with trypan blue dye using a hemocytometer.,
Soft agar assay
SiHa and HeLa cells (5 × 103 cells/ml) treated with different concentrations of PVaq (0–320 μg/ml) were mixed at 40°C with 0.35% agarose in culture medium and gelled at room temperature for 20 min over a previously gelled layer of 0.5% agarose in culture medium in 6-well plates. After incubation for 2 weeks, the colonies were counted in ten different fields using an Axiovert 200M microscope (Carl Zeiss, Germany), and the average value was calculated.,
All the results were obtained from three independent experiments, each performed in triplicates and the values have been presented as mean ± standard deviation. Differences among means were tested for statistical significance using one-way analysis of variance. The analyses were carried out using GraphPad Prism 5 software (San Diego, CA, USA). *P < 0.05, **P < 0.01, ***P < 0.001 were considered to be statistically significant.
| Results|| |
Liquid Chromatography–mass spectroscopy analysis
LCMS analysis of PVaq showed the presence of a range of phytochemicals, which included high levels of phenolics and flavonoids and moderate levels of tannins. A total of 77 compounds were identified [Supplementary [Table 1]. The peaks of the compounds were obtained at different retention times. The highest peak was at the retention time of 15.54, followed by retention times of 15.11, 10.61, 8.74, 6.95, 5.14, 4.72, 2.97, 2.83, 0.98, 0.64, and 0.45, corresponding to the compounds behenic acid (m/z 340.33413), Vogelin E (m/z 354.11033), acacetin (m/z 284.06847), leucopelargonidin 3-O-alpha-L-rhamno-beta-D-glucopyranoside (m/z 598.18977), naringenin-7-o-glucoside (m/z 434.12129), epicatechin (m/z 290.07903), proanthocyanidin b1 (m/z 578.14242), chlorogenic acid (m/z 354.09508), bergenin (m/z 328.07943), leucocynidin (m/z 306.07395), gallic acid (m/z 170.02152), mesoinositol (m/z 180.06338), caffeic acid (m/z 180.04225), and catechin (m/z 290.07903), respectively [Table 1] and [Figure 1].
|Table 1: Major compounds present in panchavalkala identified by liquid chromatography-mass spectroscopy|
Click here to view
|Figure 1: Liquid chromatography–mass spectrometry/mass spectrometry pattern of important compounds identified in aqueous extract of panchavalkala. Proanthocyanidin B1 (a), chlorogenic acid (b), epicatechin/Catechin (c), leucopelargonidin 3-O-alpha-L-rhamno-beta-D-glucopyranoside (d), naringenin-7-O-Glucoside (e), mesoinositol (f), bergenin (g), acacetin (h), gallic acid (i), leucocynidin (j), caffeic acid (k), vogelin E (l), behenic acid (m)|
Click here to view
Aqueous extract of Panchavalkala altered the viability of cervical cancer cells
The effect of PVaq was evaluated on the viability of SiHa and HeLa cells by plating them and allowing them to grow until they reached confluency. These cells were then cultured in the presence of various concentrations (0–320 μg/ml) of the extract for 24 h. There was significant reduction (P < 0.001) in the viability of SiHa (75.2% ± 12.8%) and HeLa (75.03% ± 6.06%) cells at 80 μg/ml concentration of PVaq compared to the untreated control cells [Figure 2]. IC50 of PVaq for SiHa and HeLa was found to be 125.8 and 96.0 μg/ml, respectively.
|Figure 2: Effect of aqueous extract of Panchavalkala on cell viability in SiHa and HeLa. The cells were treated with different concentrations (0–320 μg/ml) of aqueous extract of Panchavalkala for 24 h. The cell viability was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay|
Click here to view
Aqueous extract of Panchavalkala altered the growth kinetics of cancer cells
To test the effect of PVaq on the growth kinetics of SiHa and HeLa, the cells were treated with different concentrations (0, 20, 40, and 80 μg/ml) of the extract and grown for 24, 48, and 72 h. At the end of each treatment, the cells were stained with trypan blue and the number of viable cells was counted. It was observed that, compared to the control (untreated) cells, PVaq significantly (P < 0.001, n = 6) reduced the growth of SiHa and HeLa at all the tested concentrations. At the maximum concentration of 80 μg/ml, PVaq reduced the growth of SiHa by 67.4% ± 0.25%, 91.9% ± 0.1%, and 92.8% ± 0.18% at 24, 48, and 72 h, respectively [Figure 3]a. On the other hand, in HeLa, PVaq reduced the growth at 80 μg/ml by 81.7% ± 0.2%, 99.6% ± 0.05%, and 88.23% ± 0.17% at 24, 48, 72 h, respectively [Figure 3]b. This was further supported by soft agar assay wherein a dose-dependent decrease in the number of colonies was observed in both the cervical cancer cell lines [Figure 3]c. Interestingly, at 80 μg/ml concentration, PVaq significantly reduced the number of colonies in SiHa (86.9% ± 1.1%) and HeLa (82.4% ± 1.2%) compared to their respective untreated control cells [Figure 3]c. These results showed that PVaq regulated the growth of cervical cancer cells in a significant manner.
|Figure 3: Effect of aqueous extract of Panchavalkala on growth kinetics of cervical cancer cells. SiHa (a) and HeLa (b) cells were treated with aqueous extract of Panchavalkala (0–80 μg/ml) for 24–72 h and the number of viable cells were counted using the trypan blue dye exclusion method. Data represent mean ± standard deviation of three independent experiments. (c) The cervical cancer cell lines (SiHa and HeLa) (5 × 103) along with aqueous extract of Panchavalkala (0–80 μg/ml) were grown in soft agar for 2 weeks. Colonies were counted from at least ten different areas and the average of each has been plotted. The data represent mean ± standard deviation of three independent experiments|
Click here to view
| Discussion|| |
Despite therapeutic advances in cancer, the associated adverse events and chemoresistance remain major challenges to be addressed. During the past two decades, there have been extensive studies on the use of medicinal plants or their phytoconstituents as promising chemopreventive as well as anticancer agents. More than 60% of currently used anticancer drugs are originally derived from natural sources such as plants, marine organisms, and microorganisms.,
Traditional medicine that includes herbal-based drugs have been used worldwide from a long time to treat various chronic ailments including cancer. Various scientific studies, including ours, have suggested the potential of medicinal plants as anticancer drug candidates.,, Ayurveda, the main traditional medical practice in India, primarily deals with the prevention of disease through healthy food habits and lifestyle., It has been successful from very early times in using natural drugs and preventing or suppressing various tumors using various lines of treatment., Charak Samhita (1500–2000 BC), the supposed first text of Ayurveda, reports various formulations in the form of kashya (decoctions), each formulation containing different herbs that target a specific action.
Panchavalkala is a well- known Ayurvedic polyherbal formulation that has been reported to be used against inflammation, to clear ulcers, dress wounds, as a douche in leukorrhea and other vaginal diseases. The free radical scavenging activity of Panchavalkala and its individual components has been reported. Panchavalkala has been reported to be used as an adjunct in the treatment of leukorrhea. The bark of the individual components of Panchavalkala which include T. populnea Solandexcorea,, F. benghalensis L,, and F. religiosa L have been proven to possess antioxidant and anti-inflammatory activities. The present work reports for the first time the anticancer activity of Panchavalkala in cervical cancer.
PVaq showed presence of around 77 phytocompounds. Behenic acid is a fatty acid; vogelin is an isoflavone; and leucocyanidin is a flavonoid with no reported pharmacological properties. Acacetin is a flavonoid with anti-inflammatory and anticancer properties. Leucopelargonidin 3-O-alpha-L-rhamno-beta-D-glucopyranoside is a flavonoid with reported antidiabetic property. Naringenin-7-O-Glucoside, epicatechin, and proanthocyanidin B1 are flavonoids with reported antioxidant,, and antitumor,, properties. Bergenin is a glycoside with antihepatotoxic, antiulcerogenic, anti-HIV, antifungal, hepatoprotective, antiarrhythmic, neuroprotective, anti-inflammatory, immunomodulatory, and burn wound healing properties. Chlorogenic acid is a bioflavonoid which exhibits pharmacological activity such as antioxidant, antidiabetic, and antiobesity. Gallic acid is a phenol with antiviral, anti-inflammatory, anticancer, and antidiabetic properties. Mesoinositol is a sugar alcohol with no reported pharmacological activity. Caffeic acid and catechin are polyphenols with anti-inflammatory, anticancer, and antiviral, properties.
The presence of proanthocyanidin B1, chlorogenic acid, caffeic, and epicatechin acid present in PVaq could be contributed to the components of F. religiosa as reported earlier. The presence of leucopelargonidin 3-o-alpha-l-rhamno-beta-d-glucopyranoside, leucocynidin, naringenin-7-o-glucoside, and mesoinositol could be contributed to the phytocompounds reported in the bark extract of F. benghalensis.,,,, The phytochemicals catechin, vogelin E, and mesoinositol found in PVaq could be contributed to the reported compounds of F. virens., The The compounds behenic acid and bergeninin PVaq have been reported in the bark extract of F. glomerata. The phytocompounds acacetin and and gallic acid in PVaq have been reported in T. populnea.,, Thus, these data show the presence of marker compounds in the PVaq that could be used for confirming the authenticity of the formulation to avoid batch-to-batch variation.
Interestingly, PVaq exhibited anticancer activity against HPV-positive cervical cancer cell lines wherein it decreased the viability of the cells. Moreover, PVaq reduced the growth rate of cells in a time- and dose-dependent manner. The anticancer activity of PVaq could be attributed to the presence of different phytocompounds in the extract with reported anticancer activity. Proanthocyanidins, catechin,, naringenin-7-o-glucoside, and acacetin have been reported to suppress the growth of breast cancer cells. Mesoinositol also hasreported anticancer activity. We have previously reported that F. religiosa, one of the components of PVaq, induced cell cycle arrest in SiHa and apoptosis in HeLa and apoptosis in HeLa (HPV-18 positive) cells. These preliminary data warrant future in-depth studies at molecular and in vivo levels to delineate the mechanism of anticancer activity of Panchavalkala in detail.
| Conclusion|| |
Panchavalkala, an Ayurvedic formulation, was reported to exhibit anticancer activity against HPV-positive cervical cancer cell lines. The phytochemical evaluation of the PVaq has shown the presence of phytochemicals that have reported anticancer activity, thereby signifying the importance of this formulation as a prospective drug candidate in the management of cervical cancer. However, detailed experimentation is required in the future for understanding the underlying mechanism of its action.
The authors thank Director of IRSHA for his generous support and encouragement throughout the study period.
Financial support and sponsorship
This work was supported by funding from Bharat Seva Sansthan, Lucknow.
Conflicts of interest
There are no conflicts of interest.
| References|| |
Siegel RL, Miller KD, Jemal A. Cancer statistics, 2018. CA Cancer J Clin 2018;68:7-30.
Forman D, de Martel C, Lacey CJ, Soerjomataram I, Lortet-Tieulent J, Bruni L, et al
. Global burden of human papillomavirus and related diseases. Vaccine 2012;30 Suppl 5:F12-23.
Tewari KS, Sill MW, Long HJ 3rd
, Penson RT, Huang H, Ramondetta LM, et al
. Improved survival with bevacizumab in advanced cervical cancer. N Engl J Med 2014;370:734-43.
Florea AM, Büsselberg D. Cisplatin as an anti-tumor drug: Cellular mechanisms of activity, drug resistance and induced side effects. Cancers (Basel) 2011;3:1351-71.
Palatty PL, Kamble PS, Shirke M, Kamble S, Revankar M, Revankar VM. A clinical round up of the female infertility therapy amongst Indians. J Clin Diagn Res 2012;S6:1343-9.
Khadkutkar DK, Kanthi VG. Therapeutic uses of panchvalkal in different forms – A review. Ayurlog Natl J Res Ayurveda Sci 2014;2:1-5.
Joshi J, Bhatt R, Rege V, Vaidya R, Joshi B, Nadkarnii D, et al
. Use of cervical cytology, vaginal pH and colposcopy, adjuvant to clinical evaluation of ayurvedic vaginal douche, panchvalkal in leucorrhoea. J Cytol 2004;21:33-8. [Full text]
Neelam, Joshi D, Neeraj K. An ayurvedic management of vulvovaginitis during pregnancy. AYU 2007;28:5-10.
Dhiman K. Leucorrhoea in ayurvedic literature – A review. Ayurpharm Int J Ayur Alli Sci 2014;3:73-8.
Kumari CR, Jain CM, Pushplatha B, Bharathi K. Clinical evaluation of efficacy of kusthadi churna with udumbaradi taila in the management of karnini yonivyapad war to cervical erosion. Int J Ayurveda Pharma Res 2016;3:58-64.
Choudhari AS, Suryavanshi SA, Kaul-Ghanekar R. The aqueous extract of Ficus religiosa
induces cell cycle arrest in human cervical cancer cell lines SiHa (HPV-16 positive) and apoptosis in HeLa (HPV-18 positive). PLoS One 2013;8:e70127.
Chen XX, Lam KH, Chen QX, Leung GP, Tang SC, Sze SC, et al
. Ficus virens
proanthocyanidins induced apoptosis in breast cancer cells concomitantly ameliorated 5-fluorouracil induced intestinal mucositis in rats. Food Chem Toxicol 2017;110:49-61.
Mika D, Guruvayoorappan C. Experimental study on anti-tumor and anti-inflammatory effect of Thespesia populnea
phytochemical extract in mice models. Immunopharmacol Immunotoxicol 2013;35:157-63.
Sirdaarta J, Matthews B, White A, Cock IE. GC-MS and LC-MS analysis of Kakadu plum fruit extracts displaying inhibitory activity against microbial triggers of multiple sclerosis. Pharmacognosy Commun 2015;5:100-15.
Horai H, Arita M, Kanaya S, Nihei Y, Ikeda T, Suwa K, et al
. MassBank: A public repository for sharing mass spectral data for life sciences. J Mass Spectrom 2010;45:703-14.
Deshpande R, Mansara P, Kaul-Ghanekar R. Alpha-linolenic acid regulates cox2/VEGF/MAP kinase pathway and decreases the expression of HPV oncoproteins E6/E7 through restoration of p53 and Rb expression in human cervical cancer cell lines. Tumour Biol 2016;37:3295-305.
Koppikar SJ, Choudhari AS, Suryavanshi SA, Kumari S, Chattopadhyay S, Kaul-Ghanekar R, et al
. Aqueous cinnamon extract (ACE-c) from the bark of cinnamomum cassia causes apoptosis in human cervical cancer cell line (SiHa) through loss of mitochondrial membrane potential. BMC Cancer 2010;10:210.
Bhanot A, Sharma R, Noolvi MN. Natural sources as potential anti-cancer agents: A review. Int J Phytomed 2011;3:1-9.
Chandola HM. Lifestyle disorders: Ayurveda with lots of potential for prevention. Ayu 2012;33:327.
] [Full text]
Pandey MM, Rastogi S, Rawat AK. Indian traditional ayurvedic system of medicine and nutritional supplementation. Evid Based Complement Alternat Med 2013;2013:376327.
Mehta RG, Murillo G, Naithani R, Peng X. Cancer chemoprevention by natural products: How far have we come? Pharm Res 2010;27:950-61.
Das C, Ghosh G, Das D. Ayurvedic liquid dosage form asava and arista: An overview. Indian J Pharm Edu Res 2017;51:169-76.
Anandjiwala S, Bagul MS, Parabia M, Rajani M. Evaluation of free radical scavenging activity of an ayurvedic formulation, panchvalkala. Indian J Pharm Sci 2008;70:31-5.
] [Full text]
Ilavarasan R, Vasudevan M, Anbazhagan S, Venkataraman S. Antioxidant activity of Thespesia populnea
bark extracts against carbon tetrachloride-induced liver injury in rats. J Ethnopharmacol 2003;87:227-30.
Vasudevan M, Gunnam KK, Parle M. Antinociceptive and anti-inflammatory effects of Thespesia populnea
bark extract. J Ethnopharmacol 2007;109:264-70.
Sirisha N, Sreenivasulu M, Sangeeta K, Chetty CM. Antioxidant properties of Ficus species – A review. Int J PharmTech Res 2010;2:2174-82.
Thakare VN, Suralkar AA, Deshpande AD, Naik SR. Stem bark extraction of Ficus bengalensis
linn for anti-inflammatory and analgesic activity in animal models. Indian J Exp Biol 2010;48:39-45.
Charde RM, Dhongade HJ, Charde MS, Kasture AV. Evaluation of antioxidant, wound healing and anti-inflammatory activity of ethanolic extract of leaves of Ficusreligiosa
. Int J Pharm Sci Res 2010;19:73-82.
Pan MH, Lai CS, Wang YJ, Ho CT. Acacetin suppressed LPS-induced up-expression of iNOS and COX-2 in murine macrophages and TPA-induced tumor promotion in mice. Biochem Pharmacol 2006;72:1293-303.
Shen KH, Hung SH, Yin LT, Huang CS, Chao CH, Liu CL, et al
. Acacetin, a flavonoid, inhibits the invasion and migration of human prostate cancer DU145 cells via inactivation of the p38 MAPK signaling pathway. Mol Cell Biochem 2010;333:279-91.
Patel DK, Patel K, Kumar R, Gadewar M, Tahilyani V. Pharmacological and analytical aspects of bergenin: A concise report. Asian Pac J Trop Dis 2012;2:163-7.
Tsai SJ, Huang CS, Mong MC, Kam WY, Huang HY, Yin MC, et al
. Anti-inflammatory and antifibrotic effects of naringenin in diabetic mice. J Agric Food Chem 2012;60:514-21.
Yilmaz Y, Toledo RT. Major flavonoids in grape seeds and skins: Antioxidant capacity of catechin, epicatechin, and gallic acid. J Agric Food Chem 2004;52:255-60.
Plumb GW, De Pascual-Teresa S, Santos-Buelga C, Cheynier V, Williamson G. Antioxidant properties of catechins and proanthocyanidins: Effect of polymerisation, galloylation and glycosylation. Free Radic Res 1998;29:351-8.
Han X, Ren D, Fan P, Shen T, Lou H. Protective effects of naringenin-7-O-glucoside on doxorubicin-induced apoptosis in H9C2 cells. Eur J Pharmacol 2008;581:47-53.
Kürbitz C, Heise D, Redmer T, Goumas F, Arlt A, Lemke J, et al
. Epicatechin gallate and catechin gallate are superior to epigallocatechin gallate in growth suppression and anti-inflammatory activities in pancreatic tumor cells. Cancer Sci 2011;102:728-34.
King M, Chatelain K, Farris D, Jensen D, Pickup J, Swapp A, et al
. Oral squamous cell carcinoma proliferative phenotype is modulated by proanthocyanidins: A potential prevention and treatment alternative for oral cancer. BMC Complement Altern Med 2007;7:22.
Kim HS, Lim HK, Chung MW, Kim YC. Antihepatotoxic activity of bergenin, the major constituent of Mallotus japonicus
, on carbon tetrachloride-intoxicated hepatocytes. J Ethnopharmacol 2000;69:79-83.
Patel DK, Prasad SK, Kumar R, Hemalatha S. An overview on antidiabetic medicinal plants having insulin mimetic property. Asian Pac J Trop Biomed 2012;2:320-30.
Sato Y, Itagaki S, Kurokawa T, Ogura J, Kobayashi M, Hirano T, et al
. In vitro
and in vivo
antioxidant properties of chlorogenic acid and caffeic acid. Int J Pharm 2011;403:136-8.
Ong KW, Hsu A, Tan BK. Anti-diabetic and anti-lipidemic effects of chlorogenic acid are mediated by ampk activation. Biochem Pharmacol 2013;85:1341-51.
Kratz JM, Andrighetti-Fröhner CR, Kolling DJ, Leal PC, Cirne-Santos CC, Yunes RA, et al
. Anti-HSV-1 and anti-HIV-1 activity of gallic acid and pentyl gallate. Mem Inst Oswaldo Cruz 2008;103:437-42.
Deng H, Fang Y. Anti-inflammatory gallic acid and wedelolactone are G protein-coupled receptor-35 agonists. Pharmacology 2012;89:211-9.
Maurya DK, Nandakumar N, Devasagayam TP. Anticancer property of gallic acid in A549, a human lung adenocarcinoma cell line, and possible mechanisms. J Clin Biochem Nutr 2011;48:85-90.
Latha RC, Daisy P. Insulin-secretagogue, antihyperlipidemic and other protective effects of gallic acid isolated from Terminalia bellerica
roxb. In streptozotocin-induced diabetic rats. Chem Biol Interact 2011;189:112-8.
Norata GD, Marchesi P, Passamonti S, Pirillo A, Violi F, Catapano AL, et al
. Anti-inflammatory and anti-atherogenic effects of cathechin, caffeic acid and trans-resveratrol in apolipoprotein E deficient mice. Atherosclerosis 2007;191:265-71.
Wu J, Omene C, Karkoszka J, Bosland M, Eckard J, Klein CB, et al
. Caffeic acid phenethyl ester (CAPE), derived from a honeybee product propolis, exhibits a diversity of anti-tumor effects in pre-clinical models of human breast cancer. Cancer Lett 2011;308:43-53.
Suganuma M, Saha A, Fujiki H. New cancer treatment strategy using combination of green tea catechins and anticancer drugs. Cancer Sci 2011;102:317-23.
Wang GF, Shi LP, Ren YD, Liu QF, Liu HF, Zhang RJ, et al
. Anti-hepatitis B virus activity of chlorogenic acid, quinic acid and caffeic acid in vivo
and in vitro
. Antiviral Res 2009;83:186-90.
Song JM, Lee KH, Seong BL. Antiviral effect of catechins in green tea on influenza virus. Antiviral Res 2005;68:66-74.
Vikas VP, Vijay RP. Ficusbengalensis
. An Overview. Int J PharmBiol Sci 2010;1:1-11.
Subramanian PM, Misra GS. Chemical constituents of Ficus bengalensis
(part II). Pol J Pharmacol Pharm 1978;30:559-62.
The Wealth of India. A dictionary of Indian raw materials and industrial products. Counc Sci Ind Res 1999;4FG: 24-6.
Joseph B, Raj SJ. Phytopharmacological and phytochemical properties of three Ficus
species – An overview. Int J Pharma BioSci 2010;1:246-53.
Jain SJ, Khan T. Phytoconstituents from aerial roots of Ficus benghalensis
linn. Indo Am J Pharm Res 2015;5:3261-80.
Chen XX, Shi Y, Chai WM, Feng HL, Zhuang JX, Chen QX, et al
. Condensed tannins from Ficus virens
as tyrosinase inhibitors: Structure, inhibitory activity and molecular mechanism. PLoS One 2014;9:e91809.
Orabi MA, Orabi EA. Antiviral and antioxidant activities of flavonoids of Ficus virens
: Experimental and theoretical investigations. J Pharmacogn Phytochem 2016;5:120-8.
Malairajan P, Gopalakrishnan G, Narasimhan S, Kavimani S. Antiulcer activity of Ficus glomerata
. Pharm Biol 2007;45:674-7.
Nandhini US, Radhika V, Manisha S, Anusha JV. Phytochemical studies and antimicrobial compounds from fruit of Thespesia populnea
. Asian J Pharm Clin Res 2017;10:309-12.
Silva IK, Soysa P. Evaluation of phytochemical composition and antioxidant capacity of a decoction containing Adenanthera pavonina
L. And Thespesia populnea
L. Pharmacogn Mag 2011;7:193-9.
Daniel M. Medicinal Plants: Chemistry and Properties. Texas, USA: Science Publishers; 2006.
Pintha K, Yodkeeree S, Limtrakul P. Proanthocyanidin in red rice inhibits MDA-MB-231 breast cancer cell invasion via the expression control of invasive proteins. Biol Pharm Bull 2015;38:571-81.
Evacuasiany E, Ratnawati H, Liana LK, Widowati W, Maesaroh M, Mozef T, et al
. Cytotoxic and antioxidant activities of catechins in inhibiting the malignancy of breast cancer. Oxid Antioxid Med Sci 2014;3:141-6.
Xiang LP, Wang A, Ye JH, Zheng XQ, Polito CA, Lu JL, et al
. Suppressive effects of tea catechins on breast cancer. Nutrients 2016;8. pii: E458.
Akbarzadeh Z, Parvaresh F, Ghiasvand R, Miraghajani M. The effects of Naringenin on some human breast cancer cells: A systematic review. Arch Breast Cancer 2016;3:34-40.
Shim HY, Park JH, Paik HD, Nah SY, Kim DS, Han YS, et al
. Acacetin-induced apoptosis of human breast cancer MCF-7 cells involves caspase cascade, mitochondria-mediated death signaling and SAPK/JNK1/2-c-jun activation. Mol Cells 2007;24:95-104.
Lam S, McWilliams A, LeRiche J, MacAulay C, Wattenberg L, Szabo E, et al
. A phase I study of myo-inositol for lung cancer chemoprevention. Cancer Epidem.
[Figure 1], [Figure 2], [Figure 3]