Exploring the Phytochemical Profile and Biological Activities of Clerodendrum infortunatum.

Clerodendrum infortunatum (C. infortunatum), the hill glory bower, is reputed as the prodigious treasure for Indian folk medicine. The study has focused on exploring the phytochemistry and antitumor potential of the C. infortunatum root extract in vitro and in vivo. The ethyl acetate root extract has demonstrated the highest cytotoxicity in a series of nine human tumor cell lines. Further fractionation of the same has yielded seven compounds. The structures of these compounds were confirmed with spectroscopic techniques. Considering the toxicity observed with the crude extract, cytotoxicity of these compounds was further assessed in two breast carcinoma cell lines (MCF-7[ER/PR-positive HER2-negative] and MDA-MB-231 [ER/PR/HER2-negative]) and in two cervical cancer [human papilloma virus (HPV)-negative C33A and HPV-positive SiHa] cell lines. Betulinic acid (BA) was found as the active principle contributing the cytotoxic activity, and cervical cancer cell lines documented the minimum IC50 value in 24 h. In order to validate the in vitro experimental data, we have established a xenograft model of HPV-positive cervical cancer in female NOD/SCID mice treated with BA using doxorubicin as the positive control. BA treatment gradually reduced the tumor size, maintaining healthy hematological and biochemical parameters, and improved the survival rate of tumor-bearing mice considerably. Thus, our findings suggest that the C. infortunatum root extract has a promising anticancer property against HPV-positive cervical cancer and supports its usage by traditional healers for treating cervical cancer.

Pithecellobium dulce inhibits pulmonary metastasis induced by B16F10 melanoma cells in C57BL/6 via regulating EGFR/STAT/ NFκB /AKT signaling axis.

In this present study we analyzed anti-metastatic efficacy of fruit extract of Pithecellobium dulce (FPD) against B16F10 induced pulmonary metastatic model. FPD administration significantly (p < .01) reduced lung fibrosis, as evidenced by histochemical collagen analysis by Masson's trichome staining, total collagen, hexosamine, and uronic acid. Results showed that FPD treatment significantly attenuated the expression of EGFR mediated P38 and STAT1/3 expression, thus reduced NFκB mediated signaling cascade. Further, the expression of PIP3CA mediated activation of the AKT survival signaling pathway has been analyzed. Interestingly, in FPD treated group, the expression of AKT pathway has also downregulated. Further, we analyzed the downstream regulators of NFκB and AKT signaling pathways, which include, inflammatory genes (iNOS, COX2, NFκB, TGFß1, IL5, IL1ß, IFNγ, IL6, IL10, MCP1, GMCSF), anti-apoptotic genes (BCL2 and BCLXL), cell cycle regulators (cyclin D1 and Ki67), fibrosis activator (CT1α1), angiogenesis promoter (VEGF), metastatic promoter (MMP2/9, N CADH), mucin (MUC5AC), pro-apoptotic genes (Bax, CAS3 and CAS9) and metastasis inhibitors (TIMP1/2, E CADH, p53, PTEN). The expression of inflammatory, anti-apoptotic, cell cycle regulators, fibrosis activator, angiogenesis and metastasis promoter, and mucins were markedly reduced by FPD administration. Interestingly, the level of expression of anti-metastatic genes were markedly elevated in FPD administrated group. Lung histopathology further confirmed the anti-metastatic efficacy of FPD. PRACTICAL APPLICATIONS: Different parts of P. dulce has long been used as a folklore medicine against different diseases. This study demonstrated that bioactive constituents present in the fruit extract of P. dulce (FPD) significantly attenuated proliferation via regulating EGFR/STAT/NFκB/AKT signaling axis.

Pithecellobium dulce induces apoptosis and reduce tumor burden in experimental animals via regulating pro-inflammatory cytokines and anti-apoptotic gene expression.

The present study demonstrates the efficacy of fruit extract of Pithecellobium dulce (FPD) against Dalton's lymphoma ascites (DLA) cell lines in vitro and in vivo (DLA induced ascitic and solid tumor). Administration of FPD induced apoptosis in DLA cells via p53 regulation both in vitro and in vivo. Cell viability was quantified by MTT assay. Apoptotic cells were determined by qualitative (staining methods) and quantitative analysis (Annexin-propidium iodide based flow cytometry). Expression of pro-apoptotic markers (Caspase 3, Caspase 9, and Bax) were markedly elevated, while expression of anti-apoptotic proteins (Bcl 2 and Bcl XL) were downregulated in tumor cells. FPD administration effectively reduced tumor burden, increased mean survival time via modulating NF-kB, and reduced the level of proinflammatory cytokines (IL-6, IL-1ß, GM-CSF and TNF-α). Phytochemical screening of FPD by GC/MS analysis divulged the presence of several novel bioactive chemical constituents. Further, bioactive components identified from extract were evaluated for drug-like properties by Lipinski rule of five and properties. Naringenin, nootkatone, and gallic acid showed good drug-like properties and good pharmacokinetic profiles compared to other bioactive constituents in the extract.

Antioxidant-rich fraction of Amomum subulatum fruits mitigates experimental methotrexate-induced oxidative stress by regulating TNF-α, IL-1ß, and IL-6 proinflammatory cytokines.

The culinary spice Amomum subulatum was assessed for its phytochemical composition, in vitro antioxidant potential, and in vivo ameliorating effect against methotrexate (MTX)-induced toxicities. Phytochemical analysis of methanolic extract of A. subulatum dry fruits (MEAS) confirmed the presence of different bioactive secondary metabolites. MEAS scavenged reactive free radicals and inhibited lipid peroxidation in vitro. To confirm the antioxidant efficiency of MEAS, in vivo experiment was carried out in which MTX was administered to induce oxidative stress. Co-administration of MEAS reduced MTX-induced hepatic, renal, and pulmonary toxicities via significantly (p < .01) enhancing antioxidant status and reducing oxidative stress. MTX treatment significantly (p < .01) increased liver and kidney toxicity markers and increased proinflammatory cytokine (TNF-α, IL-1ß, and IL-6) levels. However, co-administration of MEAS significantly (p < .01) reduced their levels, and tissue histopathology confirmed the protective effect of MEAS in maintaining normal tissue architecture following MTX treatment. Protective effect of MEAS is accredited to the antioxidant and anti-inflammatory properties exhibited by bioactive compounds in MEAS. PRACTICAL APPLICATIONS: Amomum subulatum (Black cardamom) is a folkloric and culinary spice used for its organoleptic, nutritional, and medicinal properties. This study demonstrated the phytochemical composition and antioxidant potential of methanolic extract of A. subulatum dry fruits (MEAS). Toxicities associated with MTX therapy limit its clinical application. MEAS attenuated methotrexate-induced oxidative stress, inflammation, and associated organ damages, suggesting the possible therapeutic application of A. subulatum in reducing oxidative stress and associated diseases. Our results showed that A. subulatum is a potential functional food, which may be used for the betterment of health due to its richness in antioxidant and anti-inflammatory phytochemicals.

Traditional knowledge to clinical trials: a review on nutritional and therapeutic potential of Pithecellobium dulce.

The review describes botanical aspects, bioactive phytocompounds and pharmacological properties of different parts of Pithecellobium dulce, with special emphasis on the nutritional status of its fruits. The different parts of plant extract have been reported to possess anti-oxidant, anti-inflammatory, anti-microbial, anti-diabetic, cardio protective, anti-diarrhoeal, anti-ulcerogenic, larvicidal and ovicidal activities. Different parts of plant extracts were reported to contain several bioactive phytocompounds such as flavonoids, saponins, tannins, alkaloids etc. Natural products discovered so far served as a viable source for new drugs. Over the past few years, continued and perpetual attention of people has been paid to medicinal plants in connection with its remarkable importance in drug discovery. Plant products always remains a drug of choice for the identification of novel leads despite facing a tough competition from existing synthetic alternatives derived from combinatorial chemistry, owing to their efficacy, side effects, and safety. P. dulce is a highly acclaimed genus in traditional system of medicine because of its versatile nutraceutical and pharmacological properties. In this review we discuss in detail about nutritional and various therapeutic properties of P. dulce.

Edible Sword Bean Extract Induces Apoptosis in Cancer Cells In Vitro and Inhibits Ascites and Solid Tumor Development In Vivo.

Antitumor potential of edible sword bean (Canavalia gladiata (L.)) extract has been evaluated against Daltons lymphoma ascites (DLA) using in vitro and in vivo studies. Methanolic extraction was carried out and in vitro studies were performed against both DLA and A549, lung cancer cell lines. The results revealed that sword bean extract inhibited cell growth and induced apoptosis as evidenced by cytotoxic assay, Hoechst 33342 staining and acridine orange/ethidium bromide dual staining. In vivo studies performed on DLA induced solid as well as ascitic tumors models showed administration of sword bean extract (10 mg/kg B.wt.) could significantly inhibit ascitic and solid tumor development in mice. Therefore, our overall results revealed that C. gladiata treatment could significantly inhibit tumor development and induce apoptosis in tumor cells.

Fruit Extract of Pithecellobium dulce (FPD) ameliorates carrageenan-induced acute inflammatory responses via regulating pro-inflammatory mediators.

Unravelling the precise mechanisms underlying the anti-inflammatory action of fruit extract of Pithecellobium dulce (FPD) is quite complex. Hence the prime approach of this particular study is to unveil intriguing insights to its possible anti-inflammatory mechanisms. Anti-inflammatory effects of FPD were determined against experimentally induced acute and chronic inflammation in mice paw edema models. Administration of FPD significantly reduced the acute and chronic inflammation via regulating pro-inflammatory mediators such as pro-inflammatory cytokines (TNF-α, IL-6, and IL-1ß), Cycloxygenase 2 (COX-2), and inducible nitric oxide synthase (iNOS) compared to control group. The overall results suggest that FPD mitigates inflammation by regulating the inflammatory mediators. PRACTICAL APPLICATIONS: Fruit of Pithecellobium dulce is comestible and has been widely distributed in Asian pacific region. Non-steroidal anti-inflammatory drugs (NSAIDS) are among the most conventional treatment strategy against pain and inflammation. Although, chronic use of NSAIDS are associated with severe side effects such as gastrointestinal irritation, hepatic injury, excessive bleeding, and cardiovascular disorders. Hence identification of more effective complementary and alternative therapeutic approach from natural resources with fewer side effects could improve the quality of life of those receiving NSAIDS. Administration of fruit extract of Pithecellobium dulce ameliorates carrageenan-induced acute inflammatory responses, as evidenced by paw edema measurement, expression of antioxidant enzymes such as glutathionine, super oxide dismutase, pro-inflammatory cytokine analysis (IL-1ß, IL-6, and TNF-α), vascular permeability measurement, expression of COX-2 and iNOS. Further, confirmed the involvement of HO-1 pathway in anti-inflammatory action of FPD. The outcome of this present investigation could have a broad range of applications in alleviating inflammatory disorders.

Pithecellobium dulce fruit extract mitigates cyclophosphamide-mediated toxicity by regulating proinflammatory cytokines.

Pithecellobium dulce (Family: Fabaceae) is an edible fruit widely used in Asian-Pacific region. In the present study, we had investigated the protective effect of P. dulce fruit extract in mitigating harmful effects of the chemotherapeutic drug, cyclophosphamide (CTX). Our results showed that P. dulce treatment could significantly (p < .01) overcome CTX-induced immunosuppression accompanied with urotoxicity, hepatotoxicity, and nephrotoxicity in experimental animals. This was supported by histopathological data which proved that toxic effects of CTX in urinary bladder walls, liver, and kidney were markedly inhibited with P. dulce administration. Further, we observed significant alterations in in situ formation or release of granulocyte-macrophage colony-stimulation factor (GM-CSF) and interferon gamma (IFN ɤ) in the P. dulce treated group compared with cyclophosphamide control group. The outcome of the study could have wide range of applications in combating chemotherapy-associated malnutrition as well as in cancer drug development. PRACTICAL APPLICATIONS: CTX is a commonly used broad spectrum chemotherapeutic drug with severe side effects including immune suppression, malnutrition, urotoxicity, and nephrotoxicity. Identification of a novel immunomodulator from natural sources can resolve these side effects and could improve the quality of life of cancer patients receiving CTX as chemotherapeutic drug. In the present study, we had proved that P. dulce administration could significantly reduce CTX-induced immunotoxicity, urothelial toxicity, and nephrotoxicity. Administration of P. dulce showed a pronounced improvement in total leukocyte count, bone marrow cellularity/α-esterase activity, expression of antioxidant glutathione and cytokines (GM-CSF and INF-ɤ) compared to CTX-treated mice group. Further, histopathological analysis confirmed the protective efficacy of P. dulce against CTX-induced urothelial, hepato and kidney damage. These insights are fostering new combinational therapeutic approaches to cancer treatment.

Decalepis hamiltonii inhibits tumor progression and metastasis by regulating the inflammatory mediators and nuclear factor κB subunits.

Metastasis is an extremely complex process that is a major problem in the management of cancer. In the present study, we had evaluated the antimetastatic activity of DECALEPIS HAMILTONI: using B16F-10 melanoma-induced experimental lung metastasis in a C57BL/6 mice model. D HAMILTONI: treatment significantly ( : < .01) inhibited lung tumor nodule formation and reduced the lung collagen hydroxyproline, hexosamine, and uronic acid levels. Similarly serum sialic acid and γ-glutamyl transpeptidase levels were also significantly inhibited after D HAMILTONI: treatment. The levels of proinflammatory cytokines such as tumor necrosis factor α, interleukin (IL)-1ß, IL-6, granulocyte monocyte colony-stimulating factor, and IL-2 in the serum of these animals were significantly altered after D HAMILTONI: treatment. The serum NO level was also found to be significantly decreased after D HAMILTONI: treatment. This decreased NO level after D HAMILTONI: treatment was also accompanied by decreased inducible NO synthase and cyclooxygenase-2 expression. The study reveals that D HAMILTONI: treatment could alter proinflammatory cytokine production and could inhibit the activation and nuclear translocation of p65 and p50 subunits of nuclear factor κB in B16F-10 cells.

Protective effect of Bauhinia tomentosa on acetic acid induced ulcerative colitis by regulating antioxidant and inflammatory mediators.

Inflammatory bowel diseases (IBD), including Crohn's disease and Ulcerative colitis (UC), are life-long and recurrent disorders of the gastrointestinal tract with unknown etiology. The present study is designed to evaluate the ameliorative effect of Bauhinia tomentosa during ulcerative colitis (UC). Three groups of animals (n=6) were treated with B. tomentosa (5, 10, 20 mg/kg B.wt respectively) for 5 consecutive days before induction of UC. UC was induced by intracolonic injection of 3% acetic acid. The colonic mucosal injury was assessed by macroscopic scoring and histological examination. Furthermore, the mucosal content of lipid peroxidation (LPO), reduced glutathione (GSH), nitric oxide (NO), glutathione peroxidase (GPx) and superoxide dismutase (SOD) activity confirms that B. tomentosa could significantly inhibit colitis in a dose dependent manner. The myeloperoxidase (MPO), tumor necrosis factor (TNF-α), inducible nitric oxide synthase (iNOS) expression studies and lactate dehydrogenase (LDH) assay also supported that B. tomentosa could significantly inhibit experimental colitis. The effect was comparable to the standard drug sulfasalazine. Colonic mucosal injury parallels with the result of histological and biochemical evaluations. The extracts obtained from B. tomentosa possess active substances, which exert marked protective effects in acute experimental colitis, possibly by regulating the antioxidant and inflammatory mediators.

Effect of marine mangrove Avicennia marina (Forssk.) Vierh against acetic acid-induced ulcerative colitis in experimental mice.

Ulcerative colitis and Crohn's disease are two conditions that have many features in common and are referred as inflammatory bowel disease (IBD). Patients with IBD are predisposed to colorectal cancer. This investigation evaluates the effect of marine mangrove Avicennia marina against acetic acid-induced colitis. The treatment of A marina extract significantly decreased the colonic lipid peroxides, glutathione peroxidase, and serum nitric oxide and significantly increased the colonic and erythrocyte superoxide dismutase and glutathione levels compared with colitis control. In addition, A marina extract significantly decreased the lesion score and wet colon weight compared with colitis control. Treatment with A marina extract reflects its therapeutic activity against UC by minimal damage of colonic epithelial cells compared with colitis control during histopathologic examination. These protective role of A marina extract against UC could be attributed to the presence of higher levels of decanoic acid, diethylhydroxylamine (DEHA), pentanoic acid, pyrrolidine, 4-chlorophenyl, thiazolidinones, and arabinopyranoside (flavonoid). These findings suggest that A marina extract could be useful as a potential (natural) therapeutic agent for IBD.

Anti-metastatic effect of Biophytum sensitivum is exerted through its cytokine and immunomodulatory activity and its regulatory effect on the activation and nuclear translocation of transcription factors in B16F-10 melanoma cells.

The major clinical challenge for cancer therapy remains the eradication or prevention of metastatic process. This study was an investigation of the antimetastatic activity of Biophytum sensitivum, using B16F-10 melanoma-induced experimental lung metastasis in C57BL/6 mice. B. sensitivum treatment significantly reduced lung tumor nodule formation accompanied by reduced lung collagen hydroxyproline, hexosamine, and uronic acid levels. Serum sialic acid and gamma-glutamyl transpeptidase levels were also significantly inhibited after B. sensitivum treatment. B. sensitivum treatment down-regulated the expression of matrix metalloprotease-2 and -9 and at the same time upregulated the lung tissue inhibitor of metalloprotease-1 and -2 expression. The cytokine profile and growth factors such as interleukin-1beta, interleukin-6, tumor necrosis factor-alpha, granulocyte monocyte-colony stimulating factor, vascular endothelial growth factor, interleukin-2 and tissue inhibitor of metalloprotease-1 in the serum of these animals were markedly altered after B. sensitivum treatment. This altered level of cytokines after B. sensitivum treatment was also accompanied by enhanced natural killer cell and antibody-dependent cellular cytotoxicity. The study reveals that B. sensitivum treatment could alter proinflammatory cytokine production and could inhibit the activation and nuclear translocation of p65, p50, c-Rel subunits of nuclear factor-kappaB, and other transcription factors such as c-fos, activated transcription factor-2, and cyclic adenosine monophosphate response element-binding protein in B16F-10 melanoma cells.

Biophytum sensitivum (L.) DC inhibits tumor cell invasion and metastasis through a mechanism involving regulation of MMPs, prolyl hydroxylase, lysyl oxidase, nm23, ERK-1, ERK-2, STAT-1, and proinflammatory cytokine gene expression in metastatic lung tissue.

Biophytum sensitivum is a traditional oriental herbal medicine that is known for its immunostimulatory and antitumor effects. Tumor metastasis is the most important cause of cancer death. Although B sensitivum was shown to inhibit metastasis, the mechanism underlying this action is not well understood. In the present report, the authors had studied the effect of B sensitivum on the invasion and motility of B16F-10 melanoma cells and investigate the regulatory effect on the expression of matrix metalloproteases (MMPs), prolyl hydoxylase, lysyl oxidase, nm23, extracellular signal-regulated kinase (ERK)-1, ERK-2, signal transducer and activator of transcription (STAT)-1, and proinflammatory cytokines in metastatic tumor-bearing lungs. B sensitivum inhibited the invasion and motility of B16F-10 cells in a dose-dependent manner. B sensitivum inhibited the expression of MMP-2 and MMP-9, whereas it activated STAT-1 expression in metastatic tumor-bearing lungs. Similarly, inhibition of prolyl hydroxylase, lysyl oxidase, ERK-1, ERK-2, and vascular endothelial growth factor (VEGF) expression but activation of nm23 by B sensitivum was observed in metastatic tumor-bearing lungs. B sensitivum treatment also downregulated the expression of tumor necrosis factor-alpha, interleukin (IL)-1beta, IL-6, and granulocyte monocyte-colony stimulating factor in metastatic tumor-bearing lungs. In B16F-10 cells, B sensitivum also inhibited the production of proinflammatory cytokines. Overall, the results indicate that B sensitivum exhibits antimetastatic effects through the inhibition of invasion and motility. The results also suggest that MMPs, prolyl hydroxylase, lysyl oxidase, nm23, ERKs, VEGF, STAT, and proinflammatory cytokines are critical regulators of the B sensitivum-mediated antimetastatic effect.

Methanol extract of Biophytum sensitivum alters the cytokine profile and inhibits iNOS and COX-2 expression in LPS/Con A stimulated macrophages.

Biophytum sensitivum has been used in traditional folk medicine to treat numerous diseases. The molecular mechanism of B. sensitivum pharmacological and biochemical actions of macrophages in inflammation has not been clearly elucidated. We examined how the methanol extract of B. sensitivum regulates the production of interleukin (IL)-1beta, tumor necrosis factor (TNF)-alpha, IL-6, and nitric oxide (NO) in vitro and in vivo. The extract inhibits the production of NO and proinflammatory cytokines in lipopolysaccharide (LPS) or Concanavalin (Con) A-stimulated primary macrophages. In vitro L929 bioassay revealed the inhibition of TNF-alpha production by B. sensitivum treatment. Moreover, the extract could suppress the inducible nitric oxide synthase and cyclo-oxygenase-2 mRNA expression in LPS or Con A-stimulated macrophages. These findings provide evidence that B. sensitivum possesses potential anti-inflammatory activity and may be beneficial for the treatment of endotoxin shock or sepsis.