ABSTRACT
OBJECTIVE@#"Multi-targeting" drugs can prove fruitful to combat drug-resistance of multifactorial disease-cervical cancer. This study envisioned to reveal if Thuja homeopathic mother tincture (MT) and its bioactive component could combat human papillomavirus (HPV)-16-infected SiHa cervical cancer cells since it is globally acclaimed for HPV-mediated warts.@*METHODS@#Thuja MT was studied for its antiproliferative and antimigratory properties in SiHa cells followed by microscopic determination of reactive oxygen species (ROS) generation by 2',7'-dichlorodihydrofluorescein diacetate (DCFDA) staining and loss in mitochondrial membrane potential (MtMP) by rhodamine 123 (Rh123) staining. Apoptosis and autophagy inductions were studied by acridine orange/ethidium bromide (AO/EB) staining and immunoblot analyses of marker proteins. The bioactive component of Thuja MT detected by gas chromatography-mass spectrometry was studied for antiproliferative and antimigratory properties along with in silico prediction of its cellular targets by molecular docking and oral drug forming competency.@*RESULTS@#Thuja MT showed significant antiproliferative and antimigratory potential in SiHa cells at a 50% inhibitory concentration (IC50) of 17.3 µL/mL. An increase in DCFDA fluorescence and loss in Rh123 fluorescence prove that Thuja MT acted through the burst of ROS and loss in MtMP respectively. AO/EB-stained cells under the microscope and immunoblot analyses supported Thuja-induced cellular demise via dual pathways-apoptosis and autophagy. Immunoblots showed cleavage of caspase-3 and poly(adenosine diphosphate-ribose) polymerase-1 (PARP-1) along with upregulation of Beclin-1, microtubule-associated protein 1 light chain 3B (LC3B)-II, and p62 proteins. Hence, the apoptotic cascade followed a caspase-3-dependent pathway supported by PARP-1 cleavage, while autophagic death was Beclin-1-dependent and mediated by accumulation of LC3BII and p62 proteins. Thujone, detected as the bioactive principle of Thuja MT, showed greater anti-proliferative and anti-migratory potential at an IC50 of 77 µg/mL, along with excellent oral drug competency with the ability for gastrointestinal absorption and blood-brain-barrier permeation with nil toxicity. Molecular docking depicted thujone with the strongest affinity for mammalian target of rapamycin, phosphoinositide 3-kinase, and protein kinase B followed by B-cell lymphoma 2, murine double minute 2 and adenosine monophosphate-activated protein kinase, which might act as upstream triggers of apoptotic-autophagic crosstalk.@*CONCLUSION@#Robust "multi-targeting" anticancer potential of Thuja drug and thujone for HPV-infected cervical cancer ascertained its therapeutic efficacy for HPV infections.
Subject(s)
Animals , Female , Humans , Mice , Apoptosis , Autophagy , Beclin-1/pharmacology , Bicyclic Monoterpenes , Caspase 3 , Cell Line, Tumor , Mammals/metabolism , Molecular Docking Simulation , Papillomavirus Infections/drug therapy , Phosphatidylinositol 3-Kinases , Poly(ADP-ribose) Polymerase Inhibitors/therapeutic use , Reactive Oxygen Species/metabolism , Thuja/metabolism , Uterine Cervical Neoplasms/pathologyABSTRACT
Different concentrations of essential oil obtained from Acantholippia deserticola (Phil.ex F. Phil.) Moldenke were assessed against Chilean agricultural pests like Aleurothrixus floccosus (Maskell), Brevipalpus chilensis Baker, and Tetranychus urticae Koch. Thebioassays were carried out under laboratory conditions and both direct and residual applications were done through Potter precision spraytower test. The oil was obtained by steam distillation containing a rich fraction of alpha and beta-thujones (88.4 percent) and it shows marked toxic effects against pests. Indeed, a mortality of 82 percent and 89 percent was observed in both B. chilensis and T. urticae after 48 h whereas in A. floccosus over 97 percent of mortality was seen after 7 days. These results open the possibility to use essential oil from Acantholippia deserticola as natural pesticide.
Se evaluó el efecto a diferentes concentraciones de una solución de aceite esencial de Acantholippia deserticola sobre plagas agrícolas en Chile, tales como Aleurothrixus floccosus (Maskell), Brevipalpus chilensis Baker y Tetranychus urticae Koch. Los bioensayos fueron realizados mediante una torre de Potter en condiciones de laboratorio y las aplicaciones fueron directas y residuales. El aceite se obtuvo por hidrodestilación, el cual contenía una gran cantidad de alfa and beta-tuyonas (88.4 por ciento), mostrando marcados efectos tóxicos para A. floccosus, con un 97 por ciento de mortalidad después de 7 d y, para B. chilensis, y T. urticae, con una mortalidad de 82 por ciento y 89 por ciento respectivamente, después de 48 h. Estos resultados abren la posibilidad de usar aceite esencial de Acantholippia deserticola como pesticida natural.
Subject(s)
Mites , Oils, Volatile/pharmacology , Hemiptera , Pesticides/pharmacology , Verbenaceae/chemistry , Agriculture , Biological Assay , Monoterpenes/pharmacology , Pest Control, Biological , Toxicity TestsABSTRACT
Acantholippia deserticola (Phil.ex F. Phil.) Moldenke is a Verbenaceae that has long been used in traditional medicine in Tarapacá (Chile) as an analgesic, anti-inflammatory and aphrodisiac agent. Sincé alpha - and beta -thujone were identified as the main constituents (88.4 percent) of the essential oil from this plant, we investigated its biological properties. The results show that the essential oil from Acantholippia deserticola decreased locomotive and rearing activity compared to control group rats, including those treated with diazepam, but the essential oil had no effects on head movements or grooming. The essential oil also had significant anxiolytic and antidepressant effects. This essential oil, therefore, has sedative, anxiolytic and antidepressant actions on the rat central nervous system.
Acantholippia deserticola es una Verbenaceae de uso en la medicina tradicional como analgésico, antiinflamatorio y afrodisíaco en la región de Tarapacá, Chile. En el aceite esencial se ha identificado alfa - and beta -tuyonas como principales constituyentes (88.4 por ciento) de esta planta, que ha llevado a investigar sus propiedades biológicas. Los resultados muestran que el aceite esencial de Acantholippia deserticola disminuye la locomoción y el levantamiento en dos patas, en comparación con el grupo control, incluido el tratado por el diazepam, pero el aceite esencial no tuvo efecto sobre la sacudida de cabeza y el acicalamiento. En ambas pruebas, se observa un efecto significativo del aceite esencial en los efectos ansiolíticos y antidepresivos, lo que indica que el aceite esencial tiene actividad sedante, ansiolítica y antidepresiva en el sistema nervioso central.