ABSTRACT
ETHNOPHARMACOLOGICAL RELEVANCE: Cervical cancer is one of the most common malignancies in women that continues to be a public health problem worldwide. Human papillomavirus (HPV) infection is closely related as the causative agent of almost all cases of cervical cancer. Currently, there is no effective treatment for the persistence of HPV. Although vaccines have shown promising results in recent years, they are still a costly strategy for developing countries and have no therapeutic effect on existing infections, which is why the need arises to search for new strategies that can be used in treatment, suppressing oncogenic HPV and disease progression. Extracts of Schisandra Chinensis and Pueraria lobata have been used in traditional medicine, and it has been shown in recent years that some of their bioactive compounds have pharmacological, antioxidant, antitumor, apoptotic, and proliferation effects in HPV-positive cells. However, its mechanism of action has yet to be fully explored. AIM OF THE STUDY: The following study aimed to determine the chemical composition, antioxidant activity, and potential antiproliferative and viral oncogene effects of natural extracts of S. chinensis and P. lobata on HPV-18 positive cervical cancer cells. MATERIALS AND METHODS: The HPV-18-positive HeLa cells were treated for 24 and 48 h with the ethanolic extracts of S chinensis and P. lobata. Subsequently, cell viability was evaluated using the resazurin method, the effect on the cell cycle of the extracts (1.0, 10, and 100 µg/mL) was measured by flow cytometry, the gene of expression of the E6/E7, P53, BCL-2, and E2F-1 were determined by RT-PCR and the protein expression of p53, Ki-67, x|and Bcl-2 by immunohistochemistry. Additionally, the chemical characterization of the two extracts was carried out using LC-MS, and the total phenolics content (TPC), Total flavonoid content (TFC), and DPPH radical scavenging capacity were determined. Data were analyzed using the Mann-Whitney and Kruskal Wallis U test with GraphPad Prism 6 software. RESULTS: The natural extracts of Schisandra chinensis and Pueraria lobata induced down-regulation of E6 HPV oncogene (p<0.05) and a strong up-regulation of P53 (p<0.05), E2F-1 (p<0.05), and Bcl-2 (p<0.05) gene expression. Simultaneously, the natural extracts tend to increase the p53 protein levels and arrest the cell cycle of HeLa in the G1/S phase (p<0.05). Investigated extracts were characterized by the occurrence of bioactive lignans and isoflavones in S. chinensis and P. lobata, respectively. CONCLUSION: The extracts of S. chinensis and P. lobata within their chemical characterization mainly present lignan and isoflavone-type compounds, which are probably responsible for inhibiting the expression of the HPV E6 oncogene and inducing an increase in the expression of p53, Bcl -2 and E2F-1 producing cell cycle detection in S phase in HeLa cells. Therefore, these extracts are good candidates to continue studying their antiviral and antiproliferative potential in cells transformed by HPV.
Subject(s)
Papillomavirus Infections , Pueraria , Schisandra , Uterine Cervical Neoplasms , Humans , Female , HeLa Cells , Human Papillomavirus Viruses , Tumor Suppressor Protein p53/genetics , Uterine Cervical Neoplasms/drug therapy , Down-Regulation , Papillomavirus Infections/drug therapy , Oncogenes , Proto-Oncogene Proteins c-bcl-2 , AntioxidantsABSTRACT
ABSTRACT Extraction of effective components from Pueraria lobata has important value for skeletal muscle quality and gene expression. The improvement effect of traditional high-intensity intermittent training on skeletal muscle has not been obvious, and it is difficult to guarantee the properties of some volatiles. Based on this, this paper analyzes the effect of high-intensity intermittent training on skeletal muscle quality and gene expression in Pueraria lobata. Based on a brief summary of extraction of Pueraria lobata, status of research on the pharmaceutical components of Pueraria lobata was summarized. Different specimens of Pueraria lobata were selected as research objects, and the process of high-intensity intermittent training was designed. High-intensity intermittent training, solvent extraction and water solvent extraction were combined together to design the fixed-bed continuous extraction scheme. According to the influence of Pueraria lobata on skeletal muscle quality, the influence of intermittent training on skeletal muscle quality was analyzed. The extraction results showed that Pueraria lobata combined with high-intensity intermittent training can effectively improve the content of skeletal muscle and ensure the effective expression of skeletal muscle gene.
RESUMO A extração de componentes eficazes da Pueraria lobata tem importante valor para a qualidade músculoesquelética e para a expressão genética. O efeito da melhoria do tradicional treinamento intervalado de alta intensidade na estrutura músculoesquelética não tem sido óbvio, e é difícil garantir as propriedades de alguns voláteis. Com base nisso, este estudo analisa o efeito do treinamento intervalado de alta intensidade na qualidade músculoesquelética e na expressão genética na Pueraria lobata. Com base num breve resumo da extração da Pueraria lobata, resumiu-se o andamento das pesquisas sobre os componentes farmacêuticos da Pueraria lobata. Diferentes amostras de Pueraria lobata foram selecionadas como objeto de pesquisa, e formulou-se o processo do treinamento intervalado de alta intensidade. O treinamento intervalado de alta intensidade, a extração de solventes e a extração de solventes à base de água foram combinadas para conceber o sistema de extração contínua de leito fixo. De acordo com a influência da Pueraria lobata na qualidade músculoesquelética, analisou-se a influência do treino intervalado na qualidade músculoesquelética. Os resultados da extração mostraram que a Pueraria lobata, combinada com treino intervalado de alta intensidade, pode melhorar, de maneira eficaz, o teor músculoesquelético e garantir a expressão eficaz da expressão genética do músculoesquelético.
RESUMEN La extracción de componentes eficaces de la Pueraria lobata tiene un importante valor para la calidad músculoesquelética y para la expresión genética. El efecto de la mejora del tradicional entrenamiento intercalado de alta intensidad en la estructura músculoesquelética no ha sido obvio, y es difícil garantizar las propriedades de algunos volátiles. Basándose en eso, este estudio analiza el efecto del entrenamiento intercalado de alta intensidad en la calidad músculoesquelética y en la expresión genética en la Pueraria lobata. Basándose en un breve resumen de la extracción de la Pueraria lobata, se resumió el andamiento de las investigaciones sobre los componentes farmacéuticos de la Pueraria lobata. Diferentes muestras de Pueraria lobata fueron seleccionadas como objeto de investigación, y se formuló el proceso del entrenamiento intercalado de alta intensidad. El entrenamiento intercalado de alta intensidad, la extracción de solventes y la extracción de solventes a base de agua fueron combinadas para concebir el sistema de extracción continua de lecho fijo. De acuerdo con la influencia de la Pueraria lobata en la calidad músculoesquelética, se analizó la influencia del entrenamiento intercalado en la calidad músculoesquelética. Los resultados de la extracción mostraron que la Pueraria lobata, combinada con entrenamiento intercalado de alta intensidad, puede mejorar, de manera eficaz, el tenor músculoesquelético y garantizar la expresión eficaz de la expresión genética del músculoesquelético.
Subject(s)
Humans , Plant Extracts/administration & dosage , Gene Expression , Muscle, Skeletal/drug effects , Pueraria/chemistry , High-Intensity Interval Training/methodsABSTRACT
Pueraria mirifica is a Thai phytoestrogen-rich herb traditionally used for the treatment of menopausal symptoms. Pueraria lobata is also a phytoestrogen-rich herb traditionally used in Japan, Korea and China for the treatment of hypertension and alcoholism. We evaluated the mutagenic and antimutagenic activity of the two plant extracts using the Ames test preincubation method plus or minus the rat liver mixture S9 for metabolic activation using Salmonella typhimurium strains TA98 and TA100 as indicator strains. The cytotoxicity of the two extracts to the two S. typhimurium indicators was evaluated before the mutagenic and antimutagenic tests. Both extracts at a final concentration of 2.5, 5, 10, or 20 mg/plate exhibited only mild cytotoxic effects. The plant extracts at the concentrations of 2.5, 5 and 10 mg/plate in the presence and absence of the S9 mixture were negative in the mutagenic Ames test. In contrast, both extracts were positive in the antimutagenic Ames test towards either one or both of the tested mutagens 2-(2-furyl)-3-(5-nitro-2-furyl)-acrylamide and benzo(a)pyrene. The absence of mutagenic and the presence of anti-mutagenic activities of the two plant extracts were confirmed in rec-assays and further supported by a micronucleus test where both plant extracts at doses up to 300 mg/kg body weight (equivalent to 16 g/kg body weight plant tuberous powder) failed to exhibit significant micronucleus formation in rats. The tests confirmed the non-mutagenic but reasonably antimutagenic activities of the two plant extracts, supporting their current use as safe dietary supplements and cosmetics.