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Background of the Study: Thuja orientalis L. is an indigenous medicinal plant belonging to family Cupressaceae found in Darjeeling Himalayan region, Dooars and Tarai region of India. The plant cone has been used traditionally as medicine to treat various diseases, like bronchitis, bacterial skin infection, osteoarthritis, trigeminal neuralgia. Aim: The aim of the present study is to evaluate the plant cone for phytochemical constinuents, and in vitro antioxidant, and antimicrobial properties. Place and Duration of Study: All the experiments were done in the Department of Biotechnology, University of North Bengal, Darjeeling, West Bengal, 734013, India. Methodology: Methanolic extract of T. orientalis cone was analyzed for phytochemicals by various biochemical methods. Antioxidant properties were analyzed by in vitro assays of DPPH, ABTS, NO and H2O2 scavenging. Antibacterial property was analyzed by agar well diffusion method and antifungal assay was monitored by radial growth bioassay. Results: Methanolic extract of T. orientalis cone contained flavonoid, phenol, saponin, tannin, terpenoid and alkaloid. The extract showed significant in-vitro antioxidant and antimicrobial activity. Conclusions: The study revealed that T. orientalis cone has potential as source of antioxidant, antibacterial and antifungal agents. Our further study is directed towards the isolation, and characterization of active compound from methanolic extract and evaluation of its potentiality against high blood sugar.
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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
Las verrugas plantares son un tipo de infección causadas por el virus del papiloma humano (VPH). En muchos de los casos son muy dolorosas y difíciles de tratar; aunque generalmente desaparecen con tratamiento en unos meses, en otros casos persisten durante años. Se presenta el caso de un varón de 13 años que acudió a la oficina de farmacia tras el fracaso de un prolongado tratamiento convencional para una verruga plantar en el talón. Existen antecedentes documentados y experiencias favorables en el tratamiento homeopático de las verrugas plantares y de su utilización en farmacia. Se le pautaron Thuja occidentalis 15CH (10 gránulos semanalmente), Dulcamara 9CH (5-0-5), Antimonium crudum 15CH (5-0-5) y Nitricum acidum 15CH (5-0-5) durante 1 mes. Se observó una mejoría clínica importante al cabo de sólo 1 mes exclusivamente con tratamiento homeopático, que puede suponer una posibilidad de tratamiento eficaz para las verrugas plantares, incluso tras el fracaso de otras intervenciones convencionales.(AU)
Plantar warts are a type of infection caused by the human papillomavirus. In many cases they are very painful and difficult to treat, they usually disappear in a few months with the treatment but in several cases they will persist for years. The case of a 13 years old male is presented, who came to the pharmacy after a long and unsuccessful conventional treatment of a plantar wart on the heel of the left foot. The plantar warts have been successfully treated by homeopathic medicines in the bibliography and according to our own experience. The chosen medicines were Thuja occidentalis 15CH (10 granules weekly), Dulcamara 9CH 5 (5-0-5), Antimonium crudum 15CH (5-0-5) and Nitricum acidum 15CH (5-0-5). An important improvement was observed after 1 month of treatment. The homeopathic treatment can be an effective treatment option for plantar warts, including those where other conventional interventions have not succeeded. (AU)
Subject(s)
Humans , Warts/therapy , Antimonium Crudum , Nitri Acidum , Thuja , HomeopathyABSTRACT
Objective: To evaluate the genetic toxicity of Thuja essential oil by salmonella reversion test (AMES test) and mammal micronucleus test.Methods: TA97, TA98, TA100 and TA102 were used in AMES test to evaluate the mutagenesis of Thuja essential oil.Mouse bone marrow micronucleus test was conducted to assess the chromosome toxicity of the drug.Results: Both in S9 present and absent situations, the numbers of reverse mutation of Thuja essential oil at different doses for the four strains were all less than 1-fold of that of solvent control, and the difference had no statistical significance (P>0.05), suggesting negative mutation.The micronucleus test indicated that Thuja essential oil had no influence on the rate of mouse bone marrow micronucleus (P>0.05).Conclusion: Thuja essential oil shows no obvious genetic toxicity.
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Objective:To evaluate the acute oral toxicity , skin irritation and skin allergy of Thuja essential oil ( TEO) , and pro-vide experimental basis for the clinical use of TEO .Methods:The acute oral toxicity was measured by Horn ’ s assay .Totally 40 KM mice were divided into four groups and intragastrically administered with TEO at different dose of 21.50, 10.00, 4.64 and 2.15 g · kg-1 .After the 14-day observation, the death number and toxic manifestations were recorded and observed , and LD50 was calculated by checking the Horn's form of LD50 .The skin irritation test was performed on healthy adult white rabbits .Totally 9 rabbits were divid-ed into 3 groups randomly , and TEO at the concentration of 100%, 50%and 25%was painted on the skin of the rabbits .Edible vege-table oil was used as the negative control .The erythema and edema of the treated skin were evaluated and scored .Delayed skin hyper-sensitivity reaction was used to investigate the allergy of TEO .Totally 30 white guinea pigs were randomly divided into 3 groups:TEO group, the negative control (edible vegetable oil) and the positive group (1%2, 4-dinitrochlorobenzene).After the intracutaneous in-duction stage and local induction stage , TEO was used to activate the hypersensitive reaction .The skin response was observed and scored after the 24-hour and 48-hour activation.Results:The mice in 21.50 g · kg-1 TEO treatment group were all dead , while only a part of the mice in 10.00 and 4.64 g · kg-1 TEO treatment groups were dead , and no mice died in 2.15 g · kg-1 TEO treatment group.According to the Horn's form of LD50 , LD50 of TEO was 9.26 g · kg -1 for male mice and 7.94 g · kg -1 for female mice.The results of skin irritation test indicated the strong irritation effects of TEO .However , the irritation of TEO was reduced after the dilution , and 25%TEO showed no irritation to the skin of rabbits .The results of delayed skin hypersensitivity reaction showed obvious erythema and edema induced by 2, 4-dinitrochlorobenzene , while no obvious erythema and edema were found in TEO treated guinea pigs , indi-cating non-allergic effect of TEO .Conclusion:TEO has strong skin irritation in rabbits , while no obvious oral toxicity in mice and skin allergy in guinea pigs .
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Thuja occidentalis é largamente utilizada na forma de tinturas, sendo os flavonoides metabólitos de interesse da espécie, visto as distintas atividades farmacológicas citadas na literatura. Contudo, não há descrito método para a quantificação desses, sendo objetivo desse trabalho desenvolver um método colorimétrico direto utilizando cloreto de alumínio (AlCl3) por espectrofotometria UV, a fim de proporciona a quantificação de flavonoides totais, expresso em rutina. Para tal, avaliaram-se os seguintes parâmetros: concentração da solução hidroalcoólica (SH) (20, 40 e 70%); interferentes lipofílicos; volume da solução de AlCl3 2,5% (1, 2 e 3 mL) e proporção de droga vegetal (0,5, 1, 1,5 g). Adicionalmente, procedeuse a validação. A partir da avaliação dos interferentes lipofílicos, selecionou-se a SH 40% utilizando 1 g da droga vegetal. A cinética de complexação, demonstrou que a formação do complexo flavonoides-Al+3 é lenta, sendo o tempo de leitura ideal de 30 min no λ=403 nm, após a adição de 2 mL de AlCl3 2,5%. A rutina foi utilizada a conversão de absorvância em teor de flavonoides totais, uma vez que a espécie possui a presença majoritária de derivados quercetínicos do grupo flavonol na sua forma glicosilada. O método atendeu a todos os requisitos indicados para validação de uma matriz vegetal, consideração os fatores inerentes da espécie, possibilitando sua utilização na prática laboratorial.
Subject(s)
Flavonoids/analysis , Rutin/analysis , ThujaABSTRACT
The antioxidant activity of different extracts of Thuja orientalis was evaluated by employing three in vitro experiments namely (1, 1-Diphenyl-2-picrylhydrazyl) (DPPH), reducing power and chelating power assays. The stem powder of the plant was extracted with different solvents by maceration method in order of decreasing polarity and then partitioned (Flow Chart 1). The differential activities of the different extracts could be correlated with their respective phenolic and flavanoid contents and compared with standards L-ascorbic acid and butylated hydroxytoluene (BHT). All the extracts exhibit a remarkable concentration dependant scavenging activity. Moreover, among the different extract, methanol extract exhibited good scavenging response of 74.3%, 59.51% and 0.997% in DPPH, chelating power and reducing power assay in decreasing order of solvent polarity at maximum concentration, respectively. Studies are in progress to isolate and identify the active principle components responsible for this activity.