{"paper_id":"dcdc5825-c789-4455-8f1f-dc196b7c4342","body_text":"March-April 2024 Indian Journal of Pharmaceutical Sciences\n476\nResearch Paper\n*Address for correspondence\nE-mail: sabu@jntbgri.res.in\nNair et al.: Anti-Angiogenic Activity of Saraca asoca, Glycyrrhizia glabra and Curcuma longa\nThe development of angiogenesis plays an essential role in the growth and survival of endometriosis. The aim \nwas to determine the in vitro anti-angiogenic effect of Saraca asoca, Glycyrrhiza glabra, and Curcuma longa \nethanolic extract in chick embryos by emphasizing its total phenolic and flavonoid contents in the sample. \nIt was determined by Folin-Ciocalteu and aluminum chloride methods respectively. Anti-angiogenesis assay \nwas done in chorioallantoic membrane. The high amount of total phenol and flavonoid was obtained from all \nthree plant extracts and may be responsible for the bioactivity of the crude extract. Saraca asoca exhibits high \nphenolic content and anti-angiogenic activity followed by Curcuma longa with rich flavonoid and Glycyrrhiza \nglabra showed moderate activity for all the assays. A significant anti-angiogenic effect was observed for Saraca \nasoca (73.65+0.021) % and Curcuma longa  (67.65+0.024) % than reference drug dienogest (62.42+0.02) \n% in the study. Search for new therapeutic agents with anti-angiogenic potential rich in plant secondary \nmetabolites like phenols and flavonoids are need of the hour. The results successfully reveal that the above-\nmentioned plants used in traditional medicines could be used for treating angiogenesis-related diseases like \nendometriosis.\nKey words: Total phenol, total flavonoid, chorioallantoic membrane assay, gynecological disorder, Drabkin’s \nreagent\nEndometriosis is a gynecological disease \ncharacterized by the formation of endometrial \nglands and stroma seen in abnormal locations \nmainly outside the uterine cavity [1]. One of the key \nfeatures in the initiation and progression of this \ndisease condition is the promotion of angiogenesis \nresulting in inflammatory responses in the \nperitoneal cavity finally leading to the formation \nof endometriosis. Endometriotic lesions need a \nsufficient blood supply to be viable in their ectopic \nlocations [2]. Some of the common characteristics of \nthis condition can be related to cancer, including \nthe invasion of tissues with an uncontrolled growth \nprocess of angiogenesis and its ability to avoid \napoptosis [3]. The use of bioactive compounds in \nmedicinal plants could be a promising strategy for \nthe treatment of this disease [4].\nPlant extracts with phytoconstituents such as \nflavonoids and phenolic compounds have proven \ntheir beneficial effects by antioxidant, anti-\ninflammatory and pro-apoptotic functions in \nendometriosis disease management [5]. As the \ncurrent treatment method is limited and could \nproduce many side effects [6], plant-based medicines \ncould be an alternative treatment option with low \ncost [7]. Considering the complex pathogenesis of \nthis disease, plants with anti-angiogenic, anti-\ninflammatory, anti-proliferative and antioxidant \nproperties have been identified as promising \nadjuncts with better therapeutic effects. Saraca \nasoca  ( S. asoca ), Glycyrrhiza glabra ( G. glabra ) \nand Curcuma longa  (C. longa ) are medicinal plants \nIn Vitro Anti Angiogenetic Activity of Traditional Plants \nSaraca asoca, Glycyrrhizia glabra and Curcuma longa \nBeneficial for Endometriosis Treatment\nRESHMI NAIR, SHEHNA SHARAF1, LAKSHMI SREEDHARAN1, KIZHIYEDATH POLACHIRA SUJA1 AND K. K. SABU*\nJawaharlal Nehru Tropical Botanic Garden and Research Institute (JNTBGRI), Pacha-Palode (Research Centre of \nUniversity of Kerala), Thiruvananthapuram, Kerala 695562, 1Corporate Research and Development Centre, HLL Life-care \nLimited, Akkulam, Thiruvananthapuram, Kerala 695017, India\nAccepted 12 March 2024\nRevised 03 May 2023\nReceived 29 October 2022\nIndian J Pharm Sci 2024;86(2):476-483\nThis is an open access article distributed under the terms of the Creative \nCommons Attribution-NonCommercial-ShareAlike 3.0 License, which  \nallows others to remix, tweak, and build upon the work non-commercially,  \nas long as the author is credited and the new creations are licensed under \nthe identical terms\n\nMarch-April 2024Indian Journal of Pharmaceutical Sciences\n477\nwww.ijpsonline.com\nthat are well-mentioned in Indian classical books \nlike Bhaishajya Ratnavali and Ashtanga Hridaya \nemphasizing Ayurvedic formulations in treating \nendometriosis. \nS. asoca  is well known for its use in treating \nvarious gynecological disorders like menorrhagia \nin traditional systems of medicine. The bark of this \nplant is used in several Ayurvedic preparations like \nAsokarishta, Asoka Gritha, etc., and is considered \na uterine tonic [8,9] . Phytochemical investigation of \nthe bark revealed the presence of catechol, sterol, \ntannins, flavonoids and glycosides as the major \nbioactive compounds which are responsible for \nits beneficial effects like anti-estrogenic [10] , anti-\nprogestational, and anti-hemorrhagic activities [11]. \nG. glabra  is being used as a medicinal remedy \nfor gastritis issues, inflammatory disorders \nand skin-related problems [12] . The main active \ningredient of this plant is glycyrrhizin. Flavonoids \nfrom the root have been investigated to have \nantioxidant, antitumor, anti-inflammatory and \nantiangiogenic activities [13] . C. longa  (turmeric) \ncontains hydrophobic polyphenol curcumin as the \nmajor bioactive compound and possesses a wide \nrange of pharmaceutical activities such as anti-\nproliferative, anti-inflammatory, antioxidant and \ngrowth-suppressive properties [14] . Considering the \nmedicinal importance of these plants, the present \nstudy was designed to understand the role of these \nplant extracts in inhibiting angiogenesis.\nMATERIALS AND METHODS\nPlant materials: \nThe bark of S. asoca  (Fabaceae) and rhizome of \nC. longa  (Zingiberaceae) were obtained from the \nAyurvedic Research Centre (ARI), Poojappura, \nKerala and roots of G. glabra  (Fabaceae) from \nFoundation for Revitalisation of Local Health \nTraditions (FRLHT), Bangalore respectively. \nAll the plant materials were washed thoroughly \nwith distilled water for removing dust particles. \nAll these plant materials were oven dried at 40° \nfor 1 w and then powdered using a mixer grinder \nand stored in the dark at room temperature for \nexperiments.\nPreparation of plant extract:\nPowdered plant materials of 24 g were used for \nextraction by the Soxhlet method in 250 ml of \nethanol and concentrated under reduced pressure \nwith rotavapor (Buchi Labor Technik AG, Flawil, \nSwitzerland) R-210 set at 40°. All crude extracts \nobtained were stored at 4° until use.\nTotal phenol:\nFolin-Ciocalteu (FC) method with minor \nmodifications was used to determine the Total \nPhenolic Content (TPC) of different extracts using \ngallic acid as standard [15] . The FC reagent was mixed \nwith test extracts, mixed thoroughly and incubated \nin the dark for 5 min. 20 % sodium carbonate \nsolution was added to the solution and incubated \nfor 30 min at room temperature. The absorbance \nwas measured at 760 nm using a 1700 Shimadzu \nUltraviolet (UV)-visible spectrophotometer. The \ncalibration curve was prepared by employing \ngallic acid at a concentration of 10-60 µg/ml. The \nTPC was determined by using a linear regression \nequation from the standard gallic acid graph plot \nand was expressed as Gallic Acid Equivalent \n(GAE) per gram of the sample. The average of the \ntriplicates was analyzed. \nTotal flavonoid:\nAluminum chloride (AlCl 3) method was used to \ndetermine the Total Flavonoid Content (TFC) using \nquercetin as standard [16,17] . Briefly, the test extract \nwas mixed with 0.1 ml AlCl 3 and 0.1 ml Potassium \nacetate (CH 3CO2K), incubate at 30° for 10 min. \nThe absorbance was measured at 415 nm and TFC \nwas expressed as quercetin per gram (Quality \nAssurance Evaluator (QAE)) of the sample. The \naverage of triplicates was analyzed.\nChorio-Allantoic Membrane (CAM) assay:\nThe chicken CAM is a highly vascularized \nembryonic membrane that offers clear advantages \nto studying vascular functions. The anti-\nangiogenic activity was evaluated using the \nCAM model according to the previously reported \nmethod with minor modifications [18] . Fertilized \neggs were purchased from Kerala State Poultry \nDevelopment Corporation (KEPCO), Kerala. The \ndevelopment of the embryo was ensured with the \nhelp of an egg candler. Eggs were carefully surface \nsterilized with 70 % isopropanol and incubated \nat 37° with 80 % relative humidity. After then \nthe eggs were randomly divided to carry out the \nexperiment. Positive and negative controls used \nwere Dienogest (50 µg) and Phosphate-Buffered \nSaline (PBS) respectively. S. asoca , G. glabra , and \nC. longa  ethanolic extracts (250 µg) were the test \ngroups.\n\nwww.ijpsonline.com\nIndian Journal of Pharmaceutical Sciences\n478\nMarch-April 2024\nDrug administration:\nOn the 5 th d of incubation, control and test extracts \nof 20 µl were inoculated into the CAM of fertilized \neggs and sealed with parafilm. Tilted the egg gently \nand all eggs were kept in a vertical position with \nair sack upwards and were incubated not touching \nthe eggshell. \nDrabkin’s reagent test:\nDetermination of Hemoglobin (Hb) in the CAM \nwas performed as explained by Drabkin and \nAustin with slight modification [19] . On the 12 th d \nof incubation, the eggs were broken gently from \nthe side of the air sac and the inner content was \nremoved without disturbing the membrane. The \nCAM was scrapped and dispersed carefully into \na tube and centrifuged at 3000 rpm for 5 min. 20 \nµl of supernatant was added to 5 ml of Drabkin’s \nreagent. The reaction was incubated at room \ntemperature (37°) for 5 min. Hb level was measured \nat 546 nm in a spectrophotometer. The level of Hb \nin the sample is compared to a standard and results \nare expressed as anti-angiogenesis percentage. All \nthe tests were performed in triplicates to ensure \nthe reproducibility of the result. \nThe percentage of inhibition of angiogenesis was \ncalculated by the following equation: \n% Anti-angiogenisis=Optical Density (OD)/OD \nsample×100 \nRESULTS AND DISCUSSION\nBeing common ingredients in many Ayurvedic \nmedicines, S. asoca , G. glabra , and C. longa  \nhave already demonstrated potent health benefits \nto humans. All the extracts were rich in total \nphenolic and flavonoid compounds. TPC and TFC \nmethods are simple, inexpensive and quick in \nestimation. The content of phenol was expressed \nin terms of GAE (standard curve equation: \nY=0.0083X–0.0027, R 2= 0.9999) (fig. 1). It was \nobserved from the analysis that the bark of S. asoca  \nshowed the highest phenolic content mg GAE/g \nof 483.5+0.001, G. glabra  has 336+0.016 and C. \nlonga  has 164.14+0.002 in the sample (Table 1). \nThe result of the present investigation indicates \nthat flavonoid content was abundantly seen in \nC. longa  when compared to G. glabra , and S. \nasoca . The content of flavonoid was expressed in \nterms of standard quercetin (Y=0.0237X–0.2081, \nR2=0.9895) (fig. 2). Rhizome of C. longa  exhibited \nthe highest flavonoid content of (873+0.0186) mg \nGAE/g than roots of G. glabra  106.56+0.0267 and \nbark of S. asoca  49.81+0.028 equivalence (Table \n1). All data are presented as mean+Standard \nDeviation (SD).\nFig. 1: Standard graph of gallic acid\nTPC (mg GAE/g) TFC (mg QE/g)\nS. asoca 483.5±0.001 873±0.0186\nG. glabra 36±0.016 106.56±0.0267\nC. longa 164.14±0.002 49.81±0.028\nTABLE 1: ESTIMATION OF PHENOL AND FLAVONOID\n\nMarch-April 2024Indian Journal of Pharmaceutical Sciences\n479\nwww.ijpsonline.com\nthe impact of these plant extracts with reported \nSelective Estrogen Receptor Modulator (SERM) \nactivity on the treatment of endometriosis can also \nbe thoroughly investigated.\nThe chick embryo CAM model was used to \nassess anti-angiogenesis activity. In this study, \nwe have employed the same reliable method to \nstudy the anti-angiogenic effect of the selected \nplant extracts. The fertilized eggs were treated \nwith ethanolic extract of S. asoca , G. glabra , \nand C. longa . The anti-angiogenic potentials \nof the extract were evaluated on the 12 th d of \ntreatment. The in vitro  quantitative determination \nof Hb in blood was expressed based on the \ncyanmethemoglobin method. The intensity of the \ncolor is proportional to the Hb concentration and is \ncompared to known cyanmethemoglobin standard \nequivalence at 546 nm (standard curve equation: \nY=0.0225X–0.0024, R 2=0.9995) (fig. 3). From \nthe graph, the g per decilitre (dl) concentration of \ncontrol and test extracts was calculated. It is clear \nand evident that all the extracts show effective \nanti-angiogenic activity in CAM assay with values \n(Hb/dl) of 2 for S. asoca , 3.07 for G. glabra , 2.49 \nfor C. longa  and extract combination shows 2.7 \nwith respect to control of 7.92. The standard drug \nDienogest shows an effect of 2.91 in the assay. \nThe inhibitory potential of extracts was expressed \nas a percentage anti-angiogenesis. The study \nreveals that S. asoca  showed the highest inhibition \npercentage of 73.65+0.021 followed by C. longa  \nwith 67.65+0.024 and G. glabra  with 60.43+0.04 \nin the sample (Table 2 and fig. 4). The ethanolic \nextracts inhibited the growth of blood vessels on \nCAM were photographed and can be seen in fig. \n5A-fig. 5E.\nThe rich number of natural flavonoids and \nphenol contents in plants have positive effects \non human health as antibacterial, antiviral and \nanti-inflammatory agents [20] . They may differ in \nconcentration during their growth stages and are \nconsidered to have a very vital role in disease \nmanagement. The potential benefits of these \nsecondary metabolites derived from medicinal \nplants may act through multiple cell signaling \npathways and may exert their potential effects \nby healing diseased cells without affecting \nnormal cells [21] . S. asoca  bark extract is a rich \nsource of many polyphenols such as catechin \ngroups and the estimation of total phenol and \nflavonoids are also in agreement with the previous \nreports demonstrating the plant with a variety \nof pharmacological properties [22] . Epicatechin \nis one of the primary components of this plant. \nSince-epicatechin strongly inhibits the Na +/H+ \nexchanger, it has been hypothesized that this can \nalter the fluidity of the cytosolic plasma membrane \nand inhibits the proliferation of cancer cells [23] . It \nwas also reported that licorice also exhibits anti-\ninflammatory, antiviral, gastro protective and \nstrong estrogen-like activities due to its wide range \nof flavonoid and phenolic content in the plant [24] . \nSignificant flavonoid and phenolic content in C. \nlonga  also result in different medicinal properties \nlike anti-bacterial, anti-inflammatory, anti-\ncarcinogenic and anti-proliferative properties [25] . \nAlthough there is little evidence to support the \nuse of estrogen-progestins for endometriosis, \nthe European Society of Human Reproduction \nand Embryology (ESHRE) guidelines on \nendometriosis also noted that combined hormonal \ncontraception is frequently used as a treatment for \npain associated with endometriosis [26] . Therefore, \nFig. 2: Standard graph of quercetin\n\nwww.ijpsonline.com\nIndian Journal of Pharmaceutical Sciences\n480\nMarch-April 2024\nFig. 3: Standard graph of hemoglobin\nFig. 4: Anti-angiogenic activity of selected plant extract\nFig. 5: Inhibitory effect on angiogenesis and captured digital image of the CAM section\nDrugs Dose µg/ml Tested eggs % Anti-angiogenesis\nPBS (Negative control) - 3 -\nS. asoca 250 3 (73.65±0.021) %\nG. glabra 250 3 (60.43±0.04) %\nC. longa 250 3 (67.65±0.024) %\nDienogest 50 3 (62.42±0.02) %\nTABLE 2: ANTI-ANGIOGENIC EFFECT OF EXTRACTS\nG. glabra\nDienogest\n\nMarch-April 2024Indian Journal of Pharmaceutical Sciences\n481\nwww.ijpsonline.com\nemphasize the beneficial use of conventional drugs \nin endometriosis therapy. Further research on the \nmechanism of action of the extracts has to be \nworked out in future studies to substantiate these \nfindings on the basis of lower concentrations. The \ninvolvement of angiogenesis in preventing the \ngrowth of endometriosis has been indicated by \nseveral researches. However, many of them only \nprovide circumstantial evidence, frequently based \non the downregulation of anti-inflammatory factors \nlike VEGF levels or Matrix Metalloproteinase-9 \n(MMP-9) activity [34] . According to preclinical \ninvestigations, the antiangiogenic treatment may \nhave the largest initial impact on micro metastases \nand in general, smaller dosages of an angiogenesis \ninhibitor are needed to prevent neovascularization \nof microscopic metastases than to regress a primary \ntumour [35] .\nOur present study reveals that medicinal plants like \nS. asoca , G. glabra  and C. longa  with their major \nphenolic and flavonoid compounds may have a \nsignificant role in suppressing angiogenesis. It \nmay be presumed that the collective contribution \nof phenol and flavonoid compounds in the extract \nprovides anti-angiogenic properties. The study \nhighlighted the activity of crude extract through \nCAM assay. This technique was well accepted and \ndoes not need so much of financial and technical \nsupport to perform. Although many aspects \nare relevant to the pathophysiologic process of \nendometriosis, it is clear that angiogenesis is \nnecessary for the development and maintenance of \nectopic implants in the tissue. We have investigated \nthe antiangiogenic effect through a simple method \nand the analyzed plant extracts seem to be a \npromising therapeutic source for endometriosis \ntreatment. Further, in vivo  studies and gene \nexpression studies are needed to investigate the \nrole of these phyto metabolites for their molecular \nlevel of action.\nAcknowledgement: \nWe are gratefully acknowledging the facilities \nprovided by JNTBGRI, Palode and CRDC, HLL \nLifecare Limited, Trivandrum for supporting the \nstudy. This work was financially supported by \nICMR for an SRF fellowship to Reshmi Nair.\nConflict of interest:\nThe authors declared no conflict of interests.\nAngiogenesis can be defined as the formation \nof new blood vessels from pre-existing vascular \ntissue [27] . Onishi et al. [28]  have reported that the \npathogenesis of endometriotic lesions includes \nmajor steps like blood vessel breakdown, \nmembrane degradation, surrounding extracellular \nmatrix and new blood vessel formation leading \nto angiogenesis. In these aspects, the long-term \nsurvival and proliferation of endometriotic lesions \nare critically reliant upon adequate blood supply \nvia angiogenesis [29] . The goal of an anti-angiogenic \ntherapy strategy for endometriosis illness is \nto successfully induce conception in infertile \nindividuals while simultaneously relieving pelvic \npain. Endometriotic lesions have the ability to \ncreate cytokines and growth factors that regulates \ntheir vascularization and proliferation [30] . It \nwas also reported that the neovascularization \nof endometriotic lesions is significantly aided \nby the dominant Interleukin (IL)-1 released by \nactive peritoneal macrophages, IL-1 [31] . Estradiol \npromotes endometrial Vascular Endothelial \nCell Growth Factor (VEGF) production, and its \nlevels are associated with neovascularization and \nincreased vascular permeability during the late \nproliferative phase [32] . As endometriosis condition \nis heavily reliant on estrogen, hormonal therapies \naim to reduce endogenous ovarian estrogen \nproduction\n The CAM method provides a simple and rapid \nevaluation of the anti-angiogenic effect of the \nextracts in well-developed vascular tissue [33] . \nParadkar et al. [21]  have described it as an efficient \nand widely used model for determining anti or \npro-angiogenic activity in a sample. CAM assay \nmodel is an informative method used to study the \nprocess of endometriotic lesion formation in order \nto show that a viable endometrium is necessary \nto form an endometriotic lesion and the results \nindicate that S. asoca  and C. longa  extracts have \nstrong anti-angiogenic activity than positive \ncontrol dienogest, a potent drug currently used \nin endometriosis treatment. Thus, the studies are \npromising and clinically relevant for the therapy. \nThe relevant amount of flavonoids and phenolic \ncompounds in all the studied plants may be \nresponsible for the bioactivity of the crude \nextracts and serve as an inhibitory agent for \nangiogenesis. Since the results show a proven \nanti-angiogenic effect in CAM, these extracts \n\nwww.ijpsonline.com\nIndian Journal of Pharmaceutical Sciences\n482\nMarch-April 2024\nactivities of buckwheat ( Fagopyrum esculentum  Moench) \nhulls and flour. J Ethnopharmacol 2000;72(1-2):35-42. \n18. Ngoua-Meye-Misso RL, Ndong JD, Sima-Obiang C, Ondo JP, \nNdong-Atome GR, Ovono Abessolo F, et al.  Phytochemical \nstudies, antiangiogenic, anti-inflammatory and antioxidant \nactivities of Scyphocephalium ochocoa Warb.(Myristicaceae), \nmedicinal plant from Gabon. Clin Phytosci 2018;4:1-3.\n19. Drabkin DL, Austin JH. Spectrophotometric studies: II. \nPreparations from washed blood cells; nitric oxide hemoglobin \nand sulfhemoglobin. J Biol Chem 1935;112(1):51-65. \n20. Yuliani S, Mustofa, Partadiredja G. The neuroprotective effects \nof an ethanolic turmeric ( Curcuma longa L.) extract against \ntrimethyltin-induced oxidative stress in rats . Nutr Neurosci \n2019;22(11):797-804. \n21. Paradkar P, Dandekar S, Joshi J, Amonkar A, Vaidya A. \nAssessment of in vitro -in vivo  antimigratory and anti-\nangiogenic activity of Curcuma longa  linn., and Tinospora \ncordifolia willd. extracts in cervical cancer . Int J Pharm Sci \nRev Res 2017;42(2):87-93.\n22. Kuo WL, Huang YL, Wang ST, Ni CL, Shien BJ, Chen \nCC. Chemical constituents of Trema orientalis. J Chin Med \n2007;18(1):27-36. \n23. Matteucci E, Rizvi SI, Giampietro O. Erythrocyte sodium/\nhydrogen exchange inhibition by (−) epicatechin. Cell Biol Int \n2001;25(8):771-6.\n24. Velvizhi S, Annapurani S. Estimation of total flavonoid, \nphenolic content, and free radical scavenging potential of \nGlycyrrhiza glabra root extract. Asian J Pharm Clin Res \n2018;11(4):231-5. \n25. Sharma M, Thakur P, Saini RV , Kumar R, Torino E. Unveiling \nantimicrobial and anticancerous behavior of AuNPs and \nAgNPs moderated by rhizome extracts of Curcuma longa from \ndiverse altitudes of Himalaya. Sci Rep 2020;10(1):10934. \n26. Dunselman GA, Vermeulen N, Becker C, Calhaz-Jorge C, \nD’Hooghe T, de Bie B, et al. ESHRE guideline: Management \nof women with endometriosis . Hum Reprod 2014;29(3):400-\n12. \n27. Dai ZJ, Lu WF, Gao J, Kang HF, Ma YG, Zhang SQ, et al. \nAnti-angiogenic effect of the total flavonoids in Scutellaria \nbarbata D. Don. BMC Complement Altern Med 2013;13:150. \n28. Onishi M, Ichikawa T, Kurozumi K, Date I. Angiogenesis and \ninvasion in glioma. Brain Tumor Pathol 2011;28(1):13-24. \n29. Arablou T, Kolahdouz-Mohammadi R. Curcumin and \nendometriosis: Review on potential roles and molecular \nmechanisms. Biomed Pharmacother 2018;97:91-7. \n30. Lebovic DI, Shifren JL, Ryan IP, Mueller MD, Korn AP, \nDarney PD, et al.  Ovarian steroid and cytokine modulation \nof human endometrial angiogenesis . Hum Reprod \n2000;15(suppl_3):67-77. \n31. Lebovic DI, Bentzien F, Chao V A, Garrett EN, Meng \nYG, Taylor RN. Induction of an angiogenic phenotype in \nendometriotic stromal cell cultures by interleukin-1β . Mol \nHum Reprod 2000;6(3):269-75. \n32. Charnock-Jones DS, Macpherson AM, Archer DF, \nLeslie S, Makkink WK, Sharkey AM, et al.  The effect of \nprogestins on vascular endothelial growth factor, oestrogen \nreceptor and progesterone receptor immunoreactivity and \nendothelial cell density in human endometrium . Hum Reprod \n2000;15(suppl_3):85-95. \nREFERENCES\n1. Vallée A, Lecarpentier Y . Curcumin and endometriosis . Int J \nMol Sci 2020;21(7):2440. \n2. Lagana AS, Vitale SG, Granese R, Palmara V , Ban Frangez H, \nVrtacnik-Bokal E, et al.  Clinical dynamics of Dienogest for \nthe treatment of endometriosis: From bench to bedside. Expert \nOpin Drug Metab Toxicol 2017;13(6):593-6. \n3. Swiersz LM. Role of endometriosis in cancer and tumor \ndevelopment. Ann New York Acad Sci 2002;955(1):281-92. \n4. Balan A, Moga MA, Dima L, Dinu CG, Martinescu CC, Panait \nDE, et al.  An overview on the conservative management of \nendometriosis from a naturopathic perspective: Phytochemicals \nand medicinal plants. Plants 2021;10(3):587. \n5. Della Corte L, Noventa M, Ciebiera M, Magliarditi M, Sleiman \nZ, Karaman E, et al. Phytotherapy in endometriosis: An up-to-\ndate review. J Complement Integr Med 2020;17(3):20190084. \n6. Becker CM, Gattrell WT, Gude K, Singh SS. Reevaluating \nresponse and failure of medical treatment of endometriosis: A \nsystematic review. Fertil Steril 2017;108(1):125-36. \n7. Ashrafizaveh A, Sabouri Fard H, Azmoudeh E. Application \nof medicinal plants, acupuncture, massage therapy and \ntranscutaneous electric nerve stimulation in treatment of \nendometriosis: Review study . Iran J Obstetr Gynecol Infertil \n2019;22(5):90-100.\n8. Verma AB, Saroj A, Gautam B, Dubey C, Tripathi S. Review \non ethnobotanical importance of Saraca indica . IJPRS \n2014;3:313-21. \n9. Balekundri A, Mannur V . Quality control of the traditional \nherbs and herbal products: A review . Future J Pharm Sci \n2020;6:1-9. \n10. Yadav NK, Saini KS, Hossain Z, Omer A, Sharma C, Gayen \nJR, et al. Saraca indica bark extract shows in vitro antioxidant, \nantibreast cancer activity and does not exhibit toxicological \neffects. Oxid Med Cell Longev 2015;2015:205360. \n11. Shahid AP, Salini S, Sasidharan N, Padikkala J, Raghavamenon \nAC, Babu TD. Effect of Saraca asoca (Asoka) on estradiol-\ninduced keratinizing metaplasia in rat uterus . J Basic Clin \nPhysiol Pharmacol 2015;26(5):509-15. \n12. Sheela ML, Ramakrishna MK, Salimath BP. Angiogenic \nand proliferative effects of the cytokine VEGF in Ehrlich \nascites tumor cells is inhibited by Glycyrrhiza glabra . Int \nImmunopharmacol 2006;6(3):494-8. \n13. Nagaraj SR, Lingaraj SM, Balaraju Y , Kumar A, Salimath BP. \nMTA1 induced angiogenesis, migration and tumor growth is \ninhibited by Glycyrrhiza glabra. IOSR J Pharm 2012;2:34-43. \n14. Anand P, Kunnumakkara AB, Newman RA, Aggarwal BB. \nBioavailability of curcumin: Problems and promises . Mol \nPharm 2007;4(6):807-18.\n15. Singleton VL, Orthofer R, Lamuela-Raventós RM. Analysis of \ntotal phenols and other oxidation substrates and antioxidants \nby means of folin-ciocalteu reagent. InMethods in enzymology \n1999;299:152-78.\n16. Cedric SO, Louis-Clement OE, Joseph-Privat O, Cheikna Z, \nEdouard NE, Alfred TS. Ethnotherapy study, phytochemical \nscreening and antioxidant activity of Antrocaryon klaineanum \nPierre and Anthocleista nobilis G. Don. Med Plants Gabon. Int \nJ Adv Res 2015;3(5):812-9. \n17. Quettier-Deleu C, Gressier B, Vasseur J, Dine T, Brunet \nC, Luyckx M, et al.  Phenolic compounds and antioxidant \n\nMarch-April 2024Indian Journal of Pharmaceutical Sciences\n483\nwww.ijpsonline.com\ninflammation and anti-angiogenesis . Am J Reprod Immunol \n2012;67(6):491-7. \n35. DeMoraes ED, Fogler WE, Grant D, Wahl ML, Leeper DB, \nZrada S, et al. Recombinant human Angiostatin (rhA): A phase \nI clinical trial assessing safety, Pharmacokinetics (PK) and \nPharmacodynamics (PD). Proc Am Soc Clin Oncol 2001;20:3a.\n33. Kue CS, Tan KY , LaM ML, Lee HB. Chick embryo \nChorioallantoic Membrane (CAM): An alternative predictive \nmodel in acute toxicological studies for anti-cancer drugs. Exp \nAnim 2015;64(2):129-38. \n34. Xu Z, Zhao F, Lin F, Chen J, Huang Y . Lipoxin A 4 inhibits \nthe development of endometriosis in mice: The role of anti-","source_license":"CC0","license_restricted":false}