Exploring the Anticancer Potential of Spathodea campanulata, Ziziphus oenoplia, Blepharis maderaspatensis: A Bioactive Treasure for Future Therapeutics

Medicinal plants have been playing a significant role in the healthcare system worldwide and remain a crucial source for the discovery of new drugs. They form an integral part of traditional medicine, particularly in Asia, where they hold great cultural and therapeutic value. The exploration of plant-derived compounds has gained immense scientific interest due to their board pharmacological application, including cytotoxic and chemo preventive effects against cancer. As natural resources continues to provide promising alternative for disease management, the study of medicinal plants remains an important avenue for discovering new anticancer agents.¹

Over the past few decades, the global relevance of herbal medicine has significantly increased, impacting healthcare and international trade. Many communities, particularly in developing nations, continue to rely on medicinal plants due to their accessibility and affordability compared to western pharmaceuticals. Cancer, one of the most prevalent and fatal diseases, has prompted a continuous search for cost effective and effective treatment alternatives. Traditional medicine, particularly in regions like South Africa, has long utilized plant – based remedies to manage various illnesses, including cancer. Despite the rich history, much of the knowledge held by traditional healers remains undocumented. Scientific validation and conservation effector’s are crucial to preserving this invaluable traditional knowledge and ensuring its integration into modern healthcare.²

Cancer remains a global health challenge, characterized by the uncontrolled proliferation of cells leading to tumor formation. Conventional treatments such as chemotherapy and radiotherapy, while effective, often have severe side effects that weaken patients. The limitations of these therapies have led to an increased focus on plant based compounds with anticancer properties. Certain phytochemicals have shown potential in inhibiting tumor growth and inducing apoptosis in cancerous cells. Research continues to explore the role of natural compounds as alternative treatment for cancer, offering a promising future for more tolerable and effective therapies.³

Spathodea campanulata (African tulip tree) from biognoniaceae family is a tropical ornamental tree native to Africa and widely found in India. Various plant parts exhibit anti-inflammatory, analgesic, cytotoxic, and anticonvulsant properties due to the presence of alkaloids, tannins, flavonoids, glycosides and sterols. However, its anticancer potential remains unexplored. This was in-depth explained in this article where they have studied evaluated the anticancer activity of Spathodea campanulata leaf extracts on MCF -7 (breast cancer), HL-60 (leukaemia), and HT-29 (colon cancer), cell lines using the sulforhodamine B assay.⁴

Natural plant-based compound provide a promising alternative due to their potential efficacy with fewer adverse effect. One such plant of interest in Spathodea campanulata, known for diverse medicinal properties, this review explores its ethopharmacological significance, phytochemicistry, and therapeutic potential, emphasizing the need for scientific validation and integration into modern medicine including anticancer potential.⁵The World health organization (WHO) estimates that around three quarters of global population relies on herbal medicine. Spathodea campanulata P. Beauvais, it is widely used in traditional medicine. The phytochemical composition of Spathodea campanulata includes flavonoids, polyphenols, tannins, and glycosides, which contribute to its medicinal benefits. This plant is widely distributed across tropical and subtropical regions.⁵

However, Different parts of S. campanulata (leaf, bark, root, etc.) have been used against various types of disease. ⁶      

S. Campanulata for future research and health benefits. Databases utilized in the literature search includes Google scholar , PubChem ,Scopus ,web of science and PubMed .all plant names were checked with “ world flora online”.⁷

Its pharmacological potential has garnered increasing research interest, highlighting the importance of linking traditional knowledge with scientific validation. This review aims to provide updated information on the ethopharmacological uses and pharmacological potential, toxicity profile of Spathodea campanulata, offering insights for future research and integration into modern therapeutic approaches.

Studies highlight its diverse pharmacological properties, including antimicrobial, antidiabetic, anti-inflammatory, anticancer, and antioxidant effects. Phytochemical analysis reveals the presence of flavonoids, phenols, triterpenes, and alkaloids. While its medicinal relevance is well-supported, further research is needed to explore its full therapeutic potential.⁸

Blepharis maderaspatensis (L.) B. Heyne ex Roth, a member of the Acanthaceae family, is a medicinally significant plant known for its diverse pharmacological activities. Commonly referred to as Utinjan, Otigan, Utagan, and Chatushpatri in different regions, this plant thrives in arid environments and has been traditionally utilized in herbal medicine. Rich in bioactive phytoconstituents such as saponins, mucilage, and flavonoids—including caffeic acid, rutin, quercetin, and ferulic acid—B. maderaspatensis has been reported to possess antioxidant, anti-inflammatory, antimicrobial, and wound-healing properties. Additionally, its antihyperlipidaemic, anti-atherogenic, anti-ulcer, and wound-healing effects, particularly in diabetic wounds, highlight its therapeutic potential.⁹

Given its extensive medicinal applications, the present study aims to evaluate the in vitro cytotoxic potential of the ethanolic extract of B. maderaspatensis in a dose- and timedependent manner. The study employs the MTT assay to assess cytotoxicity across multiple cancer cell lines, including MCF7 (breast cancer), PA1 (ovarian cancer), HT29 (colon cancer), A375 (melanoma), and HepG2 (liver cancer). The determination of IC50 values at 24, 48, and 72 hours will provide insights into its cytotoxic efficacy compared to standard anticancer drugs. Understanding the cytotoxic profile of B. maderaspatensis may contribute to further investigations regarding its mechanism of action, isolation of active compounds, and potential role in anticancer drug development.¹⁰

Types of Cancer:

Cancer remains a major global health concern with various types affecting populations differently. Breast cancer is the most common, with high death rates in Africa due to poor awareness and healthcare. Cervical cancer, caused by HPV, is prevalent in Nigerian women but can be prevented through vaccination. Prostate cancer affects 1.28 million men globally, while colorectal cancer impacts 1.93 million people. Stomach cancer accounts for 1.09 million cases, and non-melanoma skin cancer is one of the most frequent, with 1.20 million cases. Childhood cancer affects around 400,000 children annually .The causes of cancer include lifestyle, environmental, and genetic factors. Tobacco use is responsible for 22% of cancer deaths, while poor diet, obesity, lack of physical activity, and excess alcohol consumption each contribute to around 10%. Pollutant exposure (e.g., pesticides) and radiation also increased risk. Genetic mutations, such as BRCA1 and BRCA2, significantly raise the likelihood of breast and ovarian cancers, especially in Nigeria. Raising awareness and promoting prevention can help reduce cancer-related deaths.¹¹

Causes of Cancer  

Cancer affects many individuals due to factors like exposure to pollutants (e.g., toxic pesticides), tobacco use (22% of deaths), poor diet, and obesity, lack of physical activity, alcohol consumption, radiation, and infections. Genetic mutations, such as BRCA1 and BRCA2, also contribute, especially in breast and ovarian cancer.Common cancers include breast (2.26 million cases), prostate (1.28 million), colorectal (1.93 million), stomach (1.09 million), and non-melanoma skin cancer (1.20 million). Childhood cancer affects around 400,000 children annually, with variations across regions.^12-15

Methods of Cancer Treatment

Early detection improves cancer survival and reduces treatment costs. Common treatments include surgery, radiotherapy, chemotherapy, and immunotherapy, depending on the cancer stage. Surgery removes tumors in early stages, often combined with radiotherapy to prevent metastasis. Advanced cases require chemotherapy, which, despite its effectiveness, lacks selectivity and causes side effects.A multidisciplinary approach using surgery, oncology, and radiotherapy is the best clinical option. However, in Nigeria, limited specialized centers make herbal medicine a preferred alternative. Traditional medicine is affordable, accessible, and has fewer side effects. Many plants contain phenolic compounds with antioxidant properties, showing potential in cancer management.^16-22

Indigenous Perspective on Cancer Treatment

In many African communities, traditional medicine plays a significant role in healthcare. According to the World Health Organization, around 80% of individuals in Africa rely on medicinal plants for treatment. Traditional medical practitioners (TMPs) have extensive knowledge of medicinal plants, their preparation, and administration, leading to successful treatment of some chronic diseases. However, a major challenge is the lack of standardization, as improper identification of plant materials can pose health risks.

Many indigenous people also avoid hospital treatment due to cultural beliefs and distrust of medical institutions. Some fear losing their cultural identity, while others believe that hospital treatments offer little chance of survival. This scepticism reinforces their reliance on traditional medicine, despite its limitations. Addressing these concerns through education and integration of traditional and modern medicine could improve cancer treatment outcomes in these communities.²³

Table: Geographical Distribution And Ethanobotanical Uses:

Table: Study Characteristics:

1. SPATHODEA CAMPANULATA:

KEY CHEMICAL CONSTITUENTS AND THEIR ACTIVITIES:  

ANTICANCER ACTIVITY:

The anticancer effects of Spathodea campanulata are mediated through multiple, wellcharacterized cellular and molecular mechanisms, attributed to its rich phytochemical  composition-especially ursolic acid, kaempferol, flavonoids, iridiodes(such as spathosides), and phenolic acids.

2. APOPTOSIS INDUCTION(Programmed cell death):

The bark of spathodea campanulata exhibits potent anticancer activity against glioma cell lines C6 and U87MG. By inducing apoptosis, it causes significant cytotoxic effects. Morphological examinations using flouresscence and confocal microscopy illustrated characteristic apoptotic features inn treated cells, such as nuclear condensation and fragmentation. The presence of

DNA fragmentation was confirmed through agarose gel electrophoresis. The extract induced apoptosis as indicated by morphological changes, DNA fragmentation, and phosphatidylserine externalization in treated glioma cells.⁶  

-(World Journal of Pharmaceutical Research, 2024)

3. CELL CYCLE ARREST:

The extract caused cell cycle arrest primarily in the G0/G1 phase, which is associated with the inhibition of glioma cell proliferation. Methods included MTT assays for cytotoxicity, flouresence microscopy for apoptosis detection, flow cytometry for cell cycle analysis, and agarose gel electrophoresis for DNA fragmentation.⁶

-(World Journal of Pharmaceutical Research)

4. SELECTIVE CYTOTOXICITY:

Spathodea campanulata extracts were not toxic to normal kidney fibroblasts(VERO). In vivo studies described in the literature support this observation, e.g., an Spathodea campanualata leaf methonolic extract was reported not to induce acute toxicity when administered orally(5004000 mg/kg) to mice. 

The stem bark methonolic extract(SsSbM) showed that highest cytotoxicity (CC50 119.03 µg/ml) against HeLa cells. Moreover, this extract excerted selective anticancer activity against all tested cancer cell libes. When tested on VERO cells, SsSbM was nontoxic upto 500µg/ml, comparision between VERO, RKO, and HeLa cells treated with SsSbM at 250µg/ml; it can be clearly seen that the VERO cells were not afftected, wheras the RKO monolayer showed layer confluency and differences in cellular morphology in comparision to the control cells, indicating a cytotoxic effect.⁵⁶

-(Bridging the Chemical Profiles and Biological Effects of Spathodea campanulata, PMC, 2022)

5. ANTI-ANGIOGENIC/ ANTI-INFLAMMATORY ACTIVITY via VEGF AND CYTOKINE DOWNREGULATION:

The ethanolic extract of spathodea campanulata root bark significantly inhibited angiogensis in experimental models. Specifiacally, it suppressed new blood vessel formation via downregulation of Vascular Endothelial Growth Factor (VEGF), a critical signal molecule driving tumor angiogensis. The supression of VEGF was demonstrated through both in vitro and in vivo(CAM assay) models, confirming that phytochemical constituents of S.campanulata impede tumor vascularization by restricting this pro-angiogensis factor.⁵⁷

S. campanulata extracts possess significant anti-inflammatory activity, including supression of pro-inflammatory cytokjnes such as TNF-α, IL-1β, and IL-6. These properties are attributed to its rich content of flavonoids, phenolics, and specific iridoids. Anti-inflammatory action is further corroborated by experimental evidence demonstrating inhibition of cyclooxygenase2(COX-2) activity and stabilization of cell membranes, which indirectlty supports cytokine level reduction.⁵⁸

-(ScienceDirect topics, Spathodea campanulata – an overview, Membrane stabilization: a possible anti-inflammatory mechanism, PubMed PMID:25145995)

Summary Table of Mechanistic Evidence:

2. ZIZIPUS OENOPLIA:

KEY CHEMICAL CONSTITUENTS AND THEIR ACTIVITIES:

ANTICANCER ACTIVITY:  

1. APOPTOSIS via MITOCHONDRIAL PATHWAY:

Betulinic acid and oleanolic acid from Zizipus oenoplia promote apoptosis via mitochondrial disruption, indicated by increased p53 and Bax, and decreased Bcl-2 in melanoma cells. DNA fragementation and annexin V-staining also confirmed apoptosis induction.⁵⁹ -(Mahapatra A, Ryall B Ng C, et al. Eur J Pharmacol. 2022;911:174561)

Flow cytometry revealed a significant increase in the sub-G1 fraction after treatment, supporting cell cycle arrest and apoptosis.⁸

-(Nahrin A, Junaid M, Afrose SS, et al. Mini Rev Med Chem. 2022;22(4):640-60)

2. CYTOSTASIS/ CELL CYCLE ARREST:

The ethanol root extract of Zizipus oenoplia, rich in flavonoids and triterpenoids, blocked cell proliferation by arresting cells in the G0/G1 phase, with parallel downregulation of cyclin D/E transcripts in human melanoma and HL-60 cells.⁸

-(Nahrin A et al., 2022)

3. ANTIOXIDANT AND ANTI-INFLAMMATORY ACTIVITY:

The antioxidant activity was attributed to high concentrations of phenolics and flavonoids, including quercetin and gallic acid, which showed marked scavenging activity in DPPH and ABTS assays. These compounds protect against oxidative DNA damage and reduce cellular inflammation, as confirmed in radical scavenging and anti-inflammatory tests.⁸ -(Nahrin A et al., 2022)

4. ANTIANGIOGENIC EFFECT:

Betulinic acid isolated from Zizipus oenoplia demonstrated significant antiangiogenic activity by inhibiting VEGF-induced tube formation and neovascularization in CAM assays, leading to reduced tumor vascularization and nutrient supply.8 59

-(Mahapatra A et al., 2022; and Nahrin et al., 2022)

Table: Mechanisms of Anti-cancer action of zizipus oenoplia

KEY CHEMICAL CONSTITUENTS AND THEIR ACTIVITIES:

ANTICANCER ACTIVITY:

1. APOPTOSIS /ROS SYNERGY:

Both ethanol extracts and silver nanoparticles(AgNP) conjugates of Blepharis made raspatensis induce apoptosis, as evidenced by DNA fragmentation, nuclear condensation, and apoptotic body formation, with the promotion of ROS overgeneration in tumor cells.

The cytotoxic effects of the ethanol extract of Blepharis maderaspatensis against colon cancer cells, as well as wound healing and antioxidant activities, have been reported.⁶⁰

-(Baskar et al., 2012;Rajasekaran et al.,2012)…

In studies on AgNPs of closely related medicinal plants, the evidence shows concentration dependent inhibition of the growth of cancer cells (MCF-7) (IC-50=12.35µg/ml). In addition, the fluorescent microscopic analysis shows activation of caspases 3 and 9 by AgNPs that cause morphological changes (AO/EB assay) in the cell membrane and cause nuclear condensation(DAPI assay) that eventually lead to apoptotic cell death(annexin V/PI assay). It was also observed that AgNPs generate reactive oxygen species(ROS) that modulate oxidative stress in MCF-7 cells.

-(oxid med cell Longev. 2020 PMC7559220)

2. SELECTIVE CYTOTOXICITY ENHANCED BY AgNPs:

The cytotoxic effects of  Blepharis maderaspatensis leaf extract is enhanced by AgNP conjugation, significantly increasing the apoptotic index in tumor cells while sparing normal fibroblasts at effective doses.

The methanolic fracation showed high content of total flavonoid, phenol and free radical scavenging activity. At 50µg/ml showed cytotoxic effects on 72hrs. These results indicate cytotoxicity dose dependent effects on cells and no cytotoxic effect seen on normal Vero cell lines. At the same time, AgNPs display efficient cytotoxicity predominantly on cancer cells, sparing non-tumorigenic cell populations. The observation was confirmed by pharmacological investigations indicating that silver nanoparticles generated by this plant extract highlighted effective apoptotic induction via ROS generation.

-(Research jounal of pharmaceutical, biological and chemical sciences 2016 7(4): studies with Blepharis maderaspatensis)

3. IMMUNOMODULATION AND ANTI-METASTATIC ACTION:

Saponins and, flavonoids in Blepharis maderspatensis extracts contribute to downregulation of tumor-promoting cytokines(e.g., TNF-α, IL-1β, IL-6) and anti-metastatic signalling by inhibiting pathways such as NF-Κb and MMPs.

Different concentrations of protocatachuic acid(10, 20, 30µg/ml) significantly suppress the

LPS- induced TNF-α, IL-6, IL-1, Inos, COX-2 proinlammatory cytokinin expressions. At 20µg/ml of protocatachuic acid shows preeminent inhibitory activity on macrophage RAW 264.7 cell line while compared with other concentrations. The isolated protocatechuic acid may facilitate inhibitory effects on LPS- induced inflammation by blocking NF-Κb translocation in nuclear and proinflammatory mediators COX-2, iNOS etc.

Table: Mechanistic Evidence For Anticancer Activity of B. Maderaspatensis: