Rauvolfia tetraphylla is an essential medicinal plant that has been widely used in traditional medicine for various disease treatments. However, the tumor suppressor activity of R. tetraphylla and its phytocompounds were not explored against triple-negative breast cancer. The current research investigated the impact of R. tetraphylla methanolic extract (RTE) and its isolated compounds Ajmaline (RTC1) and Reserpine (RTC2) on triple-negative breast cancer cell line (MDA-MB-231) focusing on anti-proliferative effects. Our study imparts that RTE and RTC2 showed promising cytotoxic effects compared to RTC1. So further experiments have proceeded with RTE and RTC2, to evaluate its proliferation, migration, and apoptotic effect. The result shows around 80% of cells were observed in the G0/G1 phase in cell cycle analysis indicating the cell cycle inhibition and duel staining clearly showed the apoptotic effect. The migration of cells after the scratch was 60.45% observed in control and 90% in treated cells showing the inhibition of migration. ROS distribution was intense compared to control indicating the increased ROS stress in treated cells. Both RTE and RTC2-treated cells showed the potential to suppress proliferation and induce apoptotic change by upregulating BAX and MST-1 and suppressing Bcl2, LATS-1, and YAP, proving that deregulation of YAP resulting in the blockage of TEAD-YAP complex and inhibit proliferation. Therefore, R. tetraphylla extract and its isolated compounds were demonstrated to find its ability to act against MDA-MB-231 and these findings will help adjudicate it as a therapeutic drug against experimental triple-negative breast cancer.
The preservation of key growth factors in the human amniotic membrane (hAM) that are involved in tissue regeneration and wound healing is the primary focus of this research work. Here, we quantified the total protein content and the major growth factors present in different sample preparations of hAM. The membrane is initially processed, dehydrated, and gamma-irradiated and subsequently subjected to histological staining, cytotoxicity assays, and total protein estimation. The ELISA method was used to quantify TGF b1, bFGF, PDGF-BB, VEGF-A, and EGF in three distinct preservation samples: tissue homogenate (AC-H), ball milled powder (AC-P), and lyophilized powder (AC-L). An in-vitro scratch assay was performed to analyse cell migration and wound healing. Higher TGF-b1 and FGF-b concentrations indicate the potential impact of HAM on re-epithelialization and granular tissue formation. For major growth factors, the quantification shows no significant differences between the samples. On treating the wound area with concentrations of 0.4 mg/ml and 0.6 mg/ml, the remaining wound area for AC-H, AC-L, and AC-P are 39.71%, 40.31%, 55.99% and 25.48%, 62.8%, and 29.65%, respectively. This indicates the presence of growth factors in the membrane promotes wound healing and facilitates cell migration and proliferation. This study provides insights into the quantity of key growth factors within the human amniotic membrane, thereby presenting the approach as a viable option for treating chronic wounds. Additionally, as lyophilization preserves more growth factors and offers greater stability and shelf life than other preservation techniques, it may be an appropriate substitute for ball milling.
The study aims to elucidate the pharmacological mechanism of Rauvolfia tetraphylla against breast cancer through a comprehensive, multi-faceted approach. This includes molecular docking, molecular dynamics, and experimental validation. Initial screening via ADME analysis and network pharmacology identified key compounds and potential targets. Protein-protein interaction (PPI) network analysis pinpointed Yes-associated protein-1 (YAP) as a crucial target. Molecular docking revealed that three compounds-ajmaline, reserpine, and serpentine-exhibited strong binding affinities with YAP, with scores of -6.5 to -6.7 kcal/mol. Molecular dynamics simulations were conducted to assess the stability of these interactions further. Experimental validation showed R. tetraphylla inhibited breast cancer cell proliferation, with an IC50 of 348.69 μg/mL, while demonstrating cytoprotective effects on Vero cells (IC50: 1056.23 μg/mL). Migration assays indicated an 88.5% reduction in cell migration, and increased ROS levels signaled elevated stress in cancer cells. Apoptosis was confirmed by AO/EtBr staining. In vivo validation in a DMBA-induced mouse model confirmed significant tumor growth inhibition, supported by changes in YAP expression and histopathological analysis. These findings highlight R. tetraphylla as a promising therapeutic candidate against breast cancer, offering insights into its mechanisms and potential for future drug development and clinical applications.