Investigation Of Molecular Mechanisms Underlying The Anti-Tumor And Anti-Angiogenic Activities Of Orthosiphon Stamineus Towards Colorectal Cancer

Teh Orthosiphon stamineus Benth. (Lamiaceae) digunakan secara meluas dalam perubatan tradisional. Kajian terbaru menunjukkan bahawa 50% ekstrak ethanolik daripada Orthosiphon stamineus (EOS) dan sebatian aktif, asid rosmarinik (RA), memaparkan kesan-kesan anti-angiogenik, anti-radang dan anti-tumor yang ketara dalam pelbagai model eksperimen. Walau bagaimanapun, mekanisme yang mendasari sifat-sifat ini tidak dinilai dengan sepenuhnya. Kajian yang dijalankan ini bertujuan untuk menilaikan lagi mekanisme molekul yang mendasari anti-tumor dan anti-angiogenik. Dalam model eksperimen penghijrahan, perkembangan dan pembentukan tiub, cerakin kedua-dua EOS dan RA aktif menyebabkan perencatan ketara terhadap fungsi sel endothelial manusia (HUVECs) yang penting bagi merangsang proses angiogenesis. Dalam kedua-dua kajian in vitro dan in vivo, penindasan besar neovaskularisasi dalam model aorta tikus, CAM dan plug matrigel juga diperhatikan. Kajian cerakin multipleks menunjukkan pengurangan faktor pertumbuhan utama bagi lata pro-angiogenik dan perkembangan tumor iaitu faktor pertumbuhan endothelial vaskular (VEGF), faktor pertumbuhan fibroblast asas (b-FGF), transformasi faktor pertumbuhan transformasi (TGF-α), faktor nekrosis tumor (TNF-β) dan interleukin-1, 2, 7. Orthosiphon stamineus Benth. (Lamiaceae) tea is widely consumed traditionally for its vast medicinal value. Recent studies revealed that 50% ethanolic extract of Orthosiphon stamineus (EOS) and its active compound, rosmarinic acid (RA), displayed significant anti-angiogenic, anti-inflammatory and anti-tumor effects in various experimental models. However, the mechanisms underlying these properties have not been fully evaluated. The present work aims to further evaluate the molecular mechanisms underlying its anti-tumour and anti-angiogenic properties. In migration, proliferation and tube formation assay, both EOS and its active RA caused significant inhibition of human endothelial cell (HUVECs) functions crucial for promotion of angiogenesis. Both in vitro and in vivo studies revealed significant suppression of neovascularisation in rat aortic ring, CAM and matrigel plug. Multiplex array studies showed reduction of key growth factors for pro-angiogenic cascade and tumor development i.e. Vascular endothelial growth factor (VEGF), basic fibroblast growth factor (b-FGF), transforming growth factor alpha (TGF-α), tumor necrosis factor (TNF-β) and interleukin-1, 2, 7

Isolation, Characterization, Crystal Structure Elucidation, and Anticancer Study of Dimethyl Cardamonin, Isolated from Syzygium campanulatum

Syzygium campanulatum Korth is an equatorial, evergreen, aboriginal shrub of Malaysia. Conventionally it has been used as a stomachic. However, in the currently conducted study dimethyl cardamonin or 2′,4′-dihydroxy-6′-methoxy-3′,5′-dimethylchalcone (DMC) was isolated from S. campanulatum Korth, leaf extract. The structural characterization of DMC was carried out by making use of various techniques including UV, IR, NMR spectral followed by LC-MS, and X-ray crystallographic techniques. For determining the purity of compound, highly effective techniques including TLC, HPLC, and melting point were used. The cytotoxicity of DMC and three different extracts of S. campanulatum was evaluated against human colon cancer cell line (HT-29) by three different assays. DMC and ethanolic extract revealed potent and dose-dependent cytotoxic activity on the cancer cell line with IC50 12.6 and 90.1 µg/mL, respectively. Quite astonishingly to our knowledge, this is the very first report on S. campanulatum as being a rich source (3.5%) of DMC, X-ray crystallography, and anticancer activity on human colon cancer cells

Anti-tumor Activity of NuvastaticTM (C5OSEW5050ESA) of Orthosiphon stamineus and Rosmarinic Acid in An Athymic Nude Mice Model of Breast Cancer

The current treatment strategies for metastatic breast cancer depend on the cancer subtype by palliating symptoms and prolonging life. However, triple-negative breast cancers have no targeted treatment available. Orthosiphon stamineus (O.s) is a traditional folk medicine plant used in South East Asia to treat many diseases. The aim of this study is to evaluate the anti-tumor activity of O.s extract formulation (ID: C5EOSEW5050ESA trademarked as NuvastaticTM) and its major component, rosmarinic acid in a breast in vivo tumor xenograft model. Human triple-negative breast cancer cells were transplanted into the mammary fat pad of 20 athymic nude mice. Fourteen days post-injection, mice were randomly assigned to four groups. C5EOSEW5050ESA (200 and 400 mg/kg/day) and rosmarinic acid (32 mg/kg/day) were administered orally. The body weight and tumor size were measured twice a week. Histopathological analyses of tumor tissues were conducted. Tumor necrosis and tumor growth were determined by hematoxylin and eosin staining. A significant reduction in tumor size and growth was found in all treatment groups. No significant difference between C5EOSEW5050ESA at 200 mg/kg and rosmarinic acid in the reduction of tumor size and necrosis. However, a more significant reduction was found in tumor growth and necrosis with 400 mg/kg of C5EOSEW5050ESA treatment as compared to other treatments. These results highlighted the anti-tumor activity of C5EOSEW5050ESA in reducing breast tumor growth in mice compared to the lower dose of C5EOSEW5050ESA and rosmarinic acid. This study provides valuable insights into using C5EOSEW5050ESA as a treatment to target triple-negative breast cancers in vivo

Isolation, Characterization, Crystal Structure Elucidation of Two Flavanones and Simultaneous RP-HPLC Determination of Five Major Compounds from Syzygium campanulatum Korth

Two flavanones named (2S)-7-Hydroxy-5-methoxy-6,8-dimethyl flavanone (1), (S)-5,7-dihydroxy-6,8-dimethyl-flavanone (2), along with known chalcone, namely, (E)-2ʹ,4ʹ- dihydroxy-6ʹ-methoxy-3ʹ,5ʹ-dimethylchalcone (3) and two triterpenoids, namely, betulinic and ursolic acids (4 and 5), were isolated from the leaves of Syzygium campanulatum Korth (Myrtaceae). The structures of compounds (1 and 2) were determined on the basis of UV-visible, FTIR, NMR spectroscopies and LC-EIMS analytical techniques. Furthermore, new, simple, precise, selective, accurate, highly sensitive, efficient and reproducible RP-HPLC method was developed and validated for the quantitative analysis of the compounds (1–5) from S. campanulatum plants of five different age. RP-HPLC method was validated in terms of specificity, linearity (r2 ≤ 0.999), precision (2.0% RSD), and recoveries (94.4%–105%). The LOD and LOQ of these compounds ranged from 0.13–0.38 and 0.10–2.23 μg·mL−1, OPEN ACCESS respectively. Anti-proliferative activity of isolated flavanones (1 and 2) and standardized extract of S. campanulatum was evaluated on human colon cancer (HCT 116) cell line. Compounds (1 and 2) and extract revealed potent and dose-dependent activity with IC50 67.6, 132.9 and 93.4 μg·mL−1, respectively. To the best of our knowledge, this is the first study on isolation, characterization, X-ray crystallographic analysis of compounds (1 and 2) and simultaneous RP-HPLC determination of five major compounds (1–5) from different age of S. campanulatum plants

<i>Orthosiphon aristatus</i> (Blume) Miq Alleviates Non-Alcoholic Fatty Liver Disease via Antioxidant Activities in C57BL/6 Obese Mice and Palmitic–Oleic Acid-Induced Steatosis in HepG2 Cells

Non-alcoholic fatty liver disease (NAFLD) is the most prevalent form of liver disease. Orthosiphon aristatus (Blume) Miq, a traditional plant in South Asia, has previously been shown to attenuate obesity and hyperglycaemic conditions. Eight weeks of feeding C57BL/6 mice with the standardized O. aristatus extract (400 mg/kg) inhibited the progression of NAFLD. Liver enzymes including alanine aminotransferase and aspartate transaminase were significantly reduced in treated mice by 74.2% ± 7.69 and 52.8% ± 7.83, respectively. Furthermore, the treated mice showed a reduction in serum levels of glucose (50% ± 5.71), insulin (70.2% ± 12.09), total cholesterol (27.5% ± 15.93), triglycerides (63.2% ± 16.5), low-density lipoprotein (62.5% ± 4.93) and atherogenic risk index relative to the negative control. Histologically, O. aristatus reversed hepatic fat accumulation and reduced NAFLD severity. Notably, our results showed the antioxidant activity of O. aristatus via increased superoxide dismutase activity and a reduction of hepatic malondialdehyde levels. In addition, the levels of serum pro-inflammatory mediators (IL-6 and TNFα) decreased, indicating anti-inflammatory activity. The aqueous, hydroethanolic and ethanolic fractions of O. aristatus extract significantly reduced intracellular fat accumulation in HepG2 cells that were treated with palmitic–oleic acid. Together, these findings suggest that antioxidant activities are the primary mechanism of action of O. aristatus underlying the anti-NAFLD effects