Isolation of flavonoids from Tagetes patula (French Marigold) flowers: Cytotoxic and oxidant activity in human cervical carcinoma cells / Aislación de flavonoides de las flores de Tagetes patula (cempasúchil francés): Actividad citotóxica y oxidante en células de carcinoma cervical humano

T agetes patula , known as French Marigold, belongs to the family Asteraceae. Human papillomavirus infection is considered one of the causes of cervical cancer. This study assessed the cytotoxic activity and intracellular oxidative capacity of compounds isolated from extract of T. patula flowers as anti - cancer cervical agents. Fraction F6 of n - butanol extract was subjected to column chromatography and HPLC - ESI - MS. The isolated compo unds of T. patula were used to examine cytotoxic activity and the production of total reactive oxygen species in SiHa and HeLa cells; the cells were also characterized using scanning electron microscopy. Patulitrin was cytotoxic to SiHa and HeLa cells. An increase in ROS production was observed at different times of treatment of cells with patuletin and patulitrin. Scanning electron microscopy showed morphological changes in SiHa and HeLa cells. Thus, compounds isolated from T. patula have great treatment p otential against cervical cancer.

Tagetes patula , conocida como cempasúchil francés, pertenece a la familia Asteraceae. La infección por el virus del papiloma humano se considera una de las causas del cáncer cervical. En este estudio, se evaluó la actividad citotóxica y la capacidad oxidativa intracelular de los compuestos aislados del extracto de las flores de T. patula como agentes anticancerígenos cervicales. La fracción F6 del ext racto de n - butanol se sometió a cromatografía en columna y HPLC - ESI - MS. Los compuestos aislados de T. patula se utilizaron para examinar la actividad citotóxica y la producción total de especies reactivas de oxígeno en las células SiHa y HeLa; las células también se caracterizaron mediante microscopía electrónica de barrido. Patulitrina resultó citotóxica para las células SiHa y HeLa. Se observó un aumento en la producción de ROS en diferentes momentos del tratamiento de las células con patuletina y patulit rina. La microscopía electrónica de barrido mostró cambios morfológicos en las células SiHa y HeLa. Por lo tanto, los compuestos aislados de T. patula tienen un gran potencial de tratamiento contra el cáncer cervical.

Dyslipidemic effect of aqueous and ethanolic extracts of raw and roasted xoconostle (Opuntia joconostle) peel / Efecto dislipidémico de extractos acuosos y etanólicos de cáscara cruda y asada de xoconostle (Opuntia joconostle)

The aim of the present study was to assess the impacts of roasting and the type of extraction solvent (ethanol or water) on the hypolipidemic act ivity of xoconostle fruit peel extracts in a tyloxapol - induced model of hyperlipidemia. Water and ethanol extracts from raw and roasted Opuntia joconostle peels were obtained to quantify the phytochemicals contained within and assess their hypolipidemic ac tivity in rats (n=5) against tyloxapol - induced dyslipidemia (400 mg/kg). The raw ethanol and water extracts, as well as the roasted water extract (200 mg/kg), showed hypolipidemic activity in the tyloxapol - treated group ( p <0.05). In contrast, the roasted s ample extracted with ethanol did not show this effect. The concentrations of phenolic compounds (39.80 mg GAE/g) and flavonoids (16.42 ± 0.14 mg QE/g) were higher in the ethanolic extracts than in the aqueous extracts. Conversely, the concentration of beta lains (115.51 ± 1.66 mg/100 g) was higher in the water extracts than in the ethanol extracts. It was concluded that the roasting process modified the concentration of some phytochemicals and their antioxidant capacity in vitro , producing a hypolipidemic ef fect in tyloxapol - induced hyperlipidemic rats

El objetivo del presente estudio fue evaluar el impacto del tostado y del tipo de disolvente de e xtracción (etanol o agua) sobre la actividad hipolipidémica de los extractos de cáscara de frutos de xoconostle en un modelo de hiperlipidemia inducido por el tyloxapol. Se obtuvieron extractos acuosos y etanólicos de cáscara cruda y asada de Opuntia jocon ostle para cuantificar los fitoquímicos que contienen y evaluar su actividad hipolipidémica en ratas (n=5) contra la dislipidemia inducida por el tyloxapol (400 mg/kg). Los extractos acuosos y etanólicos crudos, así como el extracto acuoso tostado (200 mg/ kg), mostraron actividad hipolipidémica en el grupo tratado con tiloxapol ( p <0,05). En cambio, la muestra asada y extraída con etanol no mostró este efecto. Las concentraciones de compuestos fenólicos (39,80 mg GAE/g) y flavonoides (16,42 ± 0,14 mg QE/g) f ueron mayores en los extractos etanólicos que en los acuosos. Por el contrario, la concentración de betalaínas (115,51 ± 1,66 mg/100 g) fue mayor en los extractos acuosos que en los etanólicos. Se concluyó que el proceso de asado modificó la concentración de algunos fitoquímicos y su capacidad antioxidante in vitro , produciendo un efecto hipolipidémico en ratas hiperlipidémicas inducidas por el tyloxapol.

Pharmacognostical studies of Eclipta prostrata (Linn. ) grown in Sri Lanka / Estudios farmacognósticos de Eclipta prostrate (Linn. ) cultivada en Sri Lanka

Eclipta prostrata Linn. is an annual herbaceous plant used in traditional medicine, commonly known as "Trailing Eclipta" or False Daisy in English, ' Keekirindiya ' in Sinhala and ' Kayyantakara' in Tamil. The aim of this study was to conduct a detailed pharmacognostical evaluation of E. prostrata found in Sri Lanka. Anatomical, physicochemical, phytochemical studies, and quantification of phyto - constituents were performed as per WHO guidelines. Whole plant was sequ entially extracted into solvents with different polarities. Phytochemical screening and Thin Layer Chromatography (TLC) fingerprinting were carried out. Anatomical study and powder microscopy revealed useful diagnostic features. Physicochemical parameters such as moisture content, ash values (total, acid insoluble, water soluble) and extractable matter in water and ethanol were evaluated. Phytochemical screening and TLC fingerprinting revealed the presence of different types of phyto - constituents. Alkaloid, tannin, saponin, total flavonoid and total polyphenol contents were quantified. In conclusion, pharmacognostical study aids in establishing the standardization parameters of E. prostrata found in Sri Lanka.

Eclipta prostrata Linn. es una planta herbácea anual utilizada en la medicina tradicional, comúnmente conocida como "Trailing Eclipta" o False Daisy en inglés, 'Keekirindiya' en sinhala y 'Kayyan takara' en tamil. El objetivo de este estudio fue realizar una evaluación farmacognóstica detallada de E. prostrata encontrada en Sri Lanka. Se llevaron a cabo estudios anatómicos, fisicoquímicos, fitoquímicos y cuantificación de fitoconstituyentes según l as directrices de la OMS. Se extrajo secuencialmente la planta entera en solventes con diferentes polaridades. Se realizaron pruebas de tamizaje fitoquímico y huellas dactilares de cromatografía en capa delgada (TLC). El estudio anatómico y la microscopía en polvo revelaron características diagnósticas útiles. Se evaluaron parámetros fisicoquímicos como el contenido de humedad, los valores de ceniza (total, insoluble en ácido, soluble en agua) y la materia extraíble en agua y etanol. Las pruebas de tamizaje fitoquímico y las huellas dactilares de TLC revelaron la presencia de diferentes tipos de fitoconstituyentes. Se cuantificaron los contenidos de alcaloides, taninos, saponinas, flavonoides totales y polifenoles totales. En conclusión, el estudio farmacogn óstico ayuda a establecer los parámetros de estandarización de E. prostrata encontrada en Sri Lanka.

Modulación por Passiflora edulis de triglicéridos y colesterol intracelulares en líneas celulares de cáncer colorrectal SW480 y sus derivadas metastásicas (SW620) / Modulation by Passiflora edulis of intracellular triglycerides and cholesterol in SW480 col orectal cancer cell lines and their metastatic derivatives (SW620)

Cancer cells modify lipid metabolism to proliferate, Passiflora edulis ( P. edulis ) fruit juice (ZuFru) has antitumor activity, but whether a mechanism is through modulation of cell lipids is unknown. T o establish if ZuFru modifies cholesterol and triglycerides in SW480 and SW620. ZuFru composition was studied by phytochemical march; antiproliferative activity by sulforhodamine B, cholesterol , and triglycerides by Folch method. Z ufru contains anthocyanins, flavonoids, alkaloids , and tannins. Cell lines showed differences in their growth rate ( p =0.049). At 39.6 µg/m L of ZuFru, cell viability was decreased: SW480 (45.6%) and SW620 (45.1%). In SW480, cholesterol (44.6%) and triglycerides (46.5%) decreased; In SW620, cholesterol decreased 14.8% and triglycerides increased 7%, with significant differences for both lines. A ntiproliferative activity of ZuFru could be associated with the inhibition of intracellular biosynthesis of cholesterol and triglycerides in SW480. Action mechanisms need to be further investigated.

Las células cancerosas modifican el metabolismo lipídico para proliferar; el zumo de fruta (ZuFru) de Passiflora edulis ( P. edulis ) tiene activida d antitumoral, sin embargo, se desconoce si se involucran los lípidos celulares. E stablecer si ZuFru modifica colesterol y triglicéridos en células SW480 y SW620. C omposición del ZuFru, actividad antiproliferativa, colesterol y triglicéridos. Se encontraro n antocianinas, flavonoides, alcaloides y taninos. Las líneas celulares mostraron diferencias en su tasa de crecimiento ( p =0 . 049); ZuFru 39,6 µg/ml se disminuyó la viabilidad celular; SW480 (45,6%) y SW620 (45,1%); en SW480 colesterol (44,6%) y triglicérid os (46,5%) en SW620, colesterol (14,8%) y los triglicéridos aumentaron 7%, con diferencias significativas para ambas líneas. La actividad antiproliferativa del ZuFru podría estar asociada a la inhibición de la biosíntesis intracelular de colesterol y de tr iglicéridos en SW480, pero no en SW620. Estos mecanismos de acción deben ser fuertemente investigados.

Bioactivity of Myrtus communis from the Montenegro coastline / Bioactividad de Myrtus communis del litoral de Montenegro

Although numerous studies have demonstrated the biomedical potential of Myrtus communis L., (Myrtaceae) data on myrt le from Montenegro are scarce. T o evaluate antioxidant, antimutagenic and antibacterial activity of myrtle methanolic extracts. Antioxidant activity was evaluated by measuring free radicals scavenging activity, reducing power and enzyme inhibition. The strongest scavenging activity was towards DPPH radical ( 2,2 - diphenyl - 1 - picry lhydrazyl) (IC 50 1.69 - 2.25 mg/mL) and superoxide anion (IC 50 0.56 to 0.88 mg/mL), followed by high reducing power (428 - 472 mgAA/g.DE) and inhibition of XOD (IC 50 0.308 - 0.6261mg/mL). Antimutagenic activity was evaluated in reverse mutation assays with Esche richia coli WP2 oxyR mutant IC202 and deficient in the induction of antioxidant enzymes. The myrtle extracts strongly inhibited mutagenesis induced by t - BOOH, reaching 70% at the highest concentration applied. Antimicrobial activity was examined on eight different bacterial strains. Gram - positive bacteria, S. epidermis , S. aureus and M. flavus demonstrated the highest sensitivity towards extracts (MICs 4.5 - 9 mg/mL), but significantly lower towards essential oil (MIC 0.42 - 3.32 mg/mL).

Aunque numerosos estudios han demostrado el potencial biomédico de Myrtus communis L., (Myrtaceae), los datos sobre el mirto de Montenegro son escasos. E valuar la actividad antioxidante, antimutagéni ca y antibacteriana de extractos metanólicos de mirto. La actividad antioxidante se evaluó midiendo la actividad de eliminación de radicales libres, el poder reductor y la inhibición enzimática. La actividad secuestrante más fuerte fue hacia DPPH radical ( IC 50 1.69 - 2.25 mg/mL) y radicales de anión superóxido (IC 50 0.56 a 0.88 mg/mL), seguido de alto poder reductor (428 - 472 m gAA/g.DE) e inhibición de XOD (I C 50 0,308 - 0,6261 mg/m L ). La actividad antimutagénica se evaluó en ensayos de mutación inversa con Esche richia coli WP2 oxyR mutante IC202 y deficiente en la inducción de enzimas antioxidantes. Los extractos de mirto inhibieron fuertemente la mutagénesis inducida por t - BOOH, alcanzando el 70% a la mayor concentración aplicada. La actividad antimicrobiana se examinó en octo cepas bacterianas diferentes. Las bacterias grampositivas, S. epidermis , S. aureus y M. flavus demostraron la sensibilidad más alta hacia los extractos (MIC 4.5 - 9 mg/mL), pero significativamente más baja hacia el aceite esencial (MIC 0.42 - 3 .32 mg/mL). Los resultados muestran la gran perspectiva nutrafarmacéutica de la especie montenegrina Myrtus communis .

Flavonoid Myricetin as Potent Anticancer Agent: A Possibility towards Development of Potential Anticancer Nutraceuticals / 中国结合医学杂志

Good nutrition plays a crucial role in maintaining a balanced lifestyle. The beneficial effects of nutrition have been found to counteract nutritional disturbances with the expanded use of nutraceuticals to treat and manage cardiovascular diseases, cancer, and other developmental defects over the last decade. Flavonoids are found abundantly in plant-derived foods such as fruits, vegetables, tea, cocoa, and wine. Fruits and vegetables contain phytochemicals like flavonoids, phenolics, alkaloids, saponins, and terpenoids. Flavonoids can act as anti-inflammatory, anti-allergic, anti-microbial (antibacterial, antifungal, and antiviral) antioxidant, anti-cancer, and anti-diarrheal agents. Flavonoids are also reported to upregulate apoptotic activity in several cancers such as hepatic, pancreatic, breast, esophageal, and colon. Myricetin is a flavonol which is naturally present in fruits and vegetables and has shown possible nutraceutical value. Myricetin has been portrayed as a potent nutraceutical that may protect against cancer. The focus of the present review is to present an updated account of studies demonstrating the anticancer potential of myricetin and the molecular mechanisms involved therein. A better understanding of the molecular mechanism(s) underlying its anticancer activity would eventually help in its development as a novel anticancer nutraceutical having minimal side effects.

Phytochemical profile and biological activities of Bromelia antiacantha extracts / Perfil fitoquímico e atividades biológicas de extratos de Bromelia antiacantha

Reports from popular medicine usually act as a basis for the development of new drugs from natural compounds with therapeutic actions for serious diseases and prevalence such as cancer. Bromelia antiacantha Bertol. is a species of the Bromeliaceae family, considered an unconventional food plant, found in the south and midwest regions of Brazil. Despite the high nutritional content and pharmacological potential of its fruits, few scientific studies report its biological actions. Thus, this study evaluates the phytochemical profile of aqueous and ethanol extracts obtained from B. antiacantha fruits, as well as their possible antioxidant, antitumor, and cytotoxic activities. The aqueous extract exhibited phenolic compounds and flavonoids, while ethanol extracts indicated the presence of flavonoids and coumarin in their composition, regardless of the region of collection. The ethanolic extract demonstrated a more promising antioxidant effect than the aqueous extract and also induced a significant inhibition in the viability of human cervical cancer cells of the SiHa strain. In addition, treatment with both extracts did not alter the viability of non-tumor cells of the immortalized human keratinocyte lineage (HaCaT). These results bring new data about extracts obtained from a native plant, edible and traditionally used in popular medicine, opening new perspectives for its possible therapeutic application.

Relatos da medicina popular costumam atuar como referencial para o desenvolvimento de novos fármacos a partir de moléculas naturais com ações terapêuticas para doenças de alta gravidade e prevalência como o câncer. Bromelia antiacantha Bertol. é uma espécie da família Bromeliaceae, considerada uma planta alimentícia não convencional (PANC), encontrada nas regiões sul e centro-oeste do Brasil. Apesar do alto teor nutritivo e potencial farmacológico de seus frutos, poucos estudos científicos relatam suas ações biológicas. Desta forma, este estudo avalia o perfil fitoquímico de extratos aquoso e etanólico obtidos de frutos de B. antiacantha, bem como a sua possível ação antioxidante, antitumoral e citotóxica. O extrato aquoso apresentou compostos fenólicos e flavonoides, enquanto os extratos etanólicos apontam a presença de flavonóides e cumarina em sua composição, independente da região de coleta. O extrato etanólico demonstrou efeito antioxidante mais promissor do que o extrato aquoso e também induziu uma inibição significativa na viabilidade de células humanas de câncer cervical da linhagem SiHa. Além disso, o tratamento com ambos extratos não alterou a viabilidade de células não tumorais da linhagem de queratinócitos humanos imortalizados (HaCaT). Estes dados trazem novas informações sobre extratos obtidos de uma espécie vegetal nativa, comestível e já utilizada tradicionalmente, mas abrindo novas perspectivas quanto a possíveis aplicações terapêuticas.

In vivo Gonadoprotective Effects of Myricetin on Cisplatin- Induced Testicular Damage via Suppression of TLR4/NF-kB Inflammation Pathway and Heat-Shock Response / Efectos Gonadoprotectores in vivo de la Miricetina sobre el Daño Testicular Inducido por Cisplatino Mediante la Supresión de la Vía de Inflamación TLR4/NF-kB y la Respuesta al Choque Térmico

SUMMARY: The aim of this study is to reveal the gonadoprotective effects of myricetin (MYC), which has many biological properties, on cisplatin (CP)-induced testicular damage in rats. For this purpose, 40 male Wistar albino rats were divided into 4 groups as Control (group given no treatment), MYC (group given 5 mg/kg/i.p myricetin for 7 days), CP (group given 7 mg/kg/i.p cisplatin at 7th day) and MYC + CP (group given 5 mg/kg/i.p myricetin for 7 days before 7 mg/kg/i.p cisplatin injection). After administrations, testicular tissues of animals were extracted and processed according to tissue processing protocol. Hematoxylin & Eosin staining were performed to evaluate the histopathological changes and Johnsen'sTesticular Biopsy Score (JTBS) was applied and mean seminiferous tubule diameters (MSTD) were measured to compare experimental groups in terms of histopathological changes. Moreover, TLR4, NF-kB, HSP70 and HSP90 expression levels were detected by immunohistochemical staining and the density of immunoreactivity were measured to determine the difference in the expression levels of these factors among groups. Additionally, testicular apoptosis was detected via TUNEL assay. JTBS and MSTD data were significantly lower in CP group compared to other groups and MYC administrations significantly protects testicular tissue against CP-induced damage. Moreover, TLR4, NF-kB, HSP70 and HSP90 expressions and apoptotic cells significantly increased in the CP group (p<0.05). However, MYC administrations exerted a strong gonadoprotective effect on testicular tissue in terms of these parameters in MYC+CP group (p<0.05). According to our results, we suggested that MYC can be considered as a protective agent against cisplatin-induced testicular damage.

El objetivo de este estudio es revelar los efectos gonadoprotectores de la miricetina (MYC), que tiene muchas propiedades biológicas, sobre el daño testicular inducido por cisplatino (CP) en ratas. Para este propósito, se dividieron 40 ratas albinas Wistar macho en 4 grupos: Control (grupo que no recibió tratamiento), MYC (grupo que recibió 5 mg/kg/i.p de miricetina durante 7 días), CP (grupo que recibió 7 mg/kg/i.p de cisplatino al séptimo día) y MYC + CP (grupo que recibió 5 mg/ kg/i.p de miricetina durante 7 días antes de la inyección de 7 mg/ kg/i.p de cisplatino). Después de las administraciones, se extrajeron y procesaron tejidos testiculares de animales según el protocolo de procesamiento de tejidos. Se realizó tinción con hematoxilina y eosina para evaluar los cambios histopatológicos y se aplicó la puntuación de biopsia testicular de Johnsen (JTBS) y se midieron los diámetros medios de los túbulos seminíferos (MSTD) para comparar los grupos experimentales en términos de cambios histopatológicos. Además, los niveles de expresión de TLR4, NF-kB, HSP70 y HSP90 se detectaron mediante tinción inmunohistoquímica y se midió la densidad de inmunorreactividad para determinar la diferencia en los niveles de expresión de estos factores entre los grupos. Además, se detectó apoptosis testicular mediante el ensayo TUNEL. Los datos de JTBS y MSTD fueron significativamente más bajos en el grupo CP en comparación con otros grupos y las administraciones de MYC protegen significativamente el tejido testicular contra el daño inducido por CP. Además, las expresiones de TLR4, NF-kB, HSP70 y HSP90 y las células apoptóticas aumentaron significativamente en el grupo CP (p<0,05). Sin embargo, las administraciones de MYC ejercieron un fuerte efecto gonadoprotector sobre el tejido testicular en términos de estos parámetros en el grupo MYC+CP (p<0,05). Según nuestros resultados, sugerimos que MYC puede considerarse como un agente protector contra el daño testicular inducido por cisplatino.

Optimization of ultrasound - assisted extraction of phenolic compounds and flavonoids from Hortia oreadica leaf / Optimización de la extracción asistida por ultrasonido de compuestos fenólicos y flavonoides de la hoja de Hortia oreadica

Hortia oreadica is indiscriminated used by people from Cerrado. However, vegetable raw material quality is decisive in obtaining inter mediate and final products. So, this study aimed to establish quality parameters of H. oreadica . For this, we performed the phytochemical screening of H. oreadica leaf and identified the best extractive conditions for phenolic compounds and flavonoids usin g factorial experimental design, varying the alcoholic strength, extraction temperature, and solid/liquid ratio in the ultrasound - assisted extraction method. The optimum extraction condition for phenolic compounds and flavonoids was 60% alcoholic strength, 40°C temperature, and a solid/liquid ratio of 8 mg/m L . Under this setting, the phenolic and flavonoid contents were 0.171 ± 0.002 mg/m L (predicted value = 0.165) and 0.087 ± 0.002 mg/m L (predicted value = 0.084), respectively. The optimized extraction par ameters could be upscaled to develop pharmaceutical drugs or nutraceutical products from this non - traditional plant species using an eco - friendly approach.

Hortia oreadica es utilizada indiscriminadamente por la gente del Cerrado. Sin embargo, la calidad de la materia prima vegetal es determinante en la obtención de productos intermedios y finales. Por lo tanto, este estudio tuvo como objetivo establecer parámetros de calidad de H. oreadica . Para ello, realizamos el tamizaje fitoquímico de la hoja de H. oreadica e identificamos las mejores condiciones extractivas para compuestos fenólicos y flavonoides mediante un diseño experimental factorial, variando el grado alcohólico, la temperatura de extracción y la relación sólido/líquido en el método de extracción asistido por ultrasonido. La condición óptima de extracción para compuestos fenólicos y flavonoides fue de 60% de grado alcohólico, 40°C de t emperatura y una relación sólido/líquido de 8 mg/m L . Bajo esta configuración, los contenidos de fenoles y flavonoides fueron 0,171 ± 0,002 mg/m L (valor previsto = 0,165) y 0,087 ± 0,002 mg/m L (valor previsto = 0,084), respectivamente. Los parámetros de ext racción optimizados podrían ampliarse para desarrollar fármacos o productos nutracéuticos a partir de esta especie de planta no tradicional uti lizando un enfoque ecológico .

Desarrollo inicial de un fitofármaco derivado del jugo de citrus limón (limón) para el tratamiento de crisis hipertensivas / Development of a phyto-drug derived from citrus lemon juice to treat acute blood pressure elevations

Objetivo: Obtener un fitofármaco a base de extracto seco de limón con estándares de calidad, procesamiento amigable con el medio ambiente y con antecedentes de efectividad y seguridad, evaluando sus efectos en pacientes hipertensos con frecuentes crisis hipertensivas. Métodos y resultados: Se seleccionó la población de limónes de las Regiones Metropolitana y Coquimbo con las concentraciones más altas de ácido ascórbico y flavonoides totales, respectivamente. Se utilizó liofilización para obtener jugo de limón en polvo. Sólo durante el período de estudio, se informó un caso de aumento/ crisis hipertensiva. La formulación con la concentración más alta de ácido ascórbico disminuyó la presión arterial sistólica y diastólica en 16 mmHg desde 10 minutos hasta 60 minutos. Por otro lado, la formulación con mayor concentración de flavonoides disminuyó la presión arterial en 12 a 30 mmHg desde 5 a 60 minutos. Conclusión: Se obtuvo un producto innovador como complemento al manejo de los aumentos de presión arterial. Los principios activos con mayor contribución al mecanismo antihipertensivo del jugo de limón corresponden a compuestos fenólicos, específicamente, flavonoides.

Aim: To obtain a product based on a dry extract of standardized phytodrug lemon with high quality standards, using an environmentally friendly process; to evaluate its effects in hypertensive patients suffering frequent episodes of hypertensive crisis. Methods and Results: Lemons with high concentration of ascorbic acid (Coquimbo Region) or total flavonoids (Metropolitan Region) were selected. Lyophilization was used to obtain juice powder. During the study period only one case of hypertensive crisis was reported. The formula with the highest concentration of ascorbic acid decreased systolic and diastolic blood pressure by 16 mmHg from 10 minutes up to 60 minutes. On the other hand, the formula with the highest concentration of flavonoids decreased blood pressure from 12 to 30 mmHg between 5 and 60 minutes. Conclusion: An novel product to help decrease acutely elevated blood pressure was obtained that can be used as a complement to the management of acute blood pressure rises. The active principles with greater contribution to the antihypertensive mechanism of lemon juice corresponded to phenolic compounds, specifically flavonoids.

Screening for acetylcholinesterase inhibition, lipid peroxidation inhibition and antioxidant activity of medicinal plants from Morocco / Detección de inhibición de acetilcolinesterasa, inhibición de peroxidación de lípidos y actividad antioxidante de plantas medicinales de Marruecos

Acetylcholinesterase (AChE), hydrolyzes acetylcholine to choline and acetate, thereby terminating this neurotransmitter effect at cholinergic synapses. Therefore, AChE inhibition is used for counterbalance the cholinergic deficit in Alzheimer's disease (AD) patients. In the present work, in order to find new plant acetylcholinesterase inhibitors, the hydroalcoholic extracts from seventeen medicinal plant species were screened for their acetylcholinesterase inhibition activity, as well as total phenolic (TPC) and flavonoids contents (TFC) and antioxidant activity using ORAC (Oxygen Radical Absorbance Capacity) assay, and their ability to inhibit lipid peroxidation. The results revealed that Rumex acetosa, Taraxacum officinale and Hypericum perforatum extracts possessing the highest TPC and TFC, were the most effective in terms of ORAC antioxidant activity, and acetylcholinesterase inhibition, in addition to their ability to inhibit liposomes peroxidation, suggesting that those plant species may provide a substantial source of secondary metabolites, which act as natural antioxidants and acetylcholinesterase inhibitors, and may be beneficial in the treatment of AD.

La acetilcolinesterasa (AChE) hidroliza la acetilcolina se hidroliza en colina y acetato, terminando así este efecto neurotransmisor en las sinapsis colinérgicas. Por lo tanto, la inhibición de la AChE se utiliza para contrarrestar el déficit colinérgico en pacientes con enfermedad de Alzheimer (EA). En el presente trabajo, con el fin de encontrar nuevos inhibidores de la acetilcolinesterasa vegetal, se analizaron los extractos hidroalcohólicos de diecisiete especies de plantas medicinales para determinar su actividad inhibidora de la acetilcolinesterasa, así como el contenido total de fenólicos (TPC) y flavonoides (TFC) y la actividad antioxidante utilizando ORAC (Capacidad de absorbancia de radicales de oxígeno) y su capacidad para inhibir la peroxidación de lípidos. Los resultados revelaron que los extractos de Rumexacetosa, Taraxacum officinale e Hypericum perforatum que poseen los más altos TPC y TFC, fueron los más efectivos en términos de actividad antioxidante ORAC e inhibición de acetilcolinesterasa, además de su capacidad para inhibir la peroxidación de los liposomas, sugiriendo que esas especies de plantas puede proporcionar una fuente sustancial de metabolitos secundarios, que actúan como antioxidantes naturales e inhibidores de la acetilcolinesterasa, y puede ser beneficioso en el tratamiento de la EA.

Site-directed mutagenesis enhances the activity of benzylidene acetone synthase of polyketide synthase from Polygonum cuspidatum / 生物工程学报

Polygonum cuspidatum polyketide synthase 1 (PcPKS1) has the catalytic activity of chalcone synthase (CHS) and benzylidene acetone synthase (BAS), which can catalyze the production of polyketides naringenin chalcone and benzylidene acetone, and then catalyze the synthesis of flavonoids or benzylidene acetone. In this study, three amino acid sites (Thr133, Ser134, Ser33) that may affect the function of PcPKS1 were identified by analyzing the sequences of PcPKS1, the BAS from Rheum palmatum and the CHS from Arabidopsis thaliana, as well as the conformation of the catalytic site of the enzyme. Molecular modification of PcPKS1 was carried out by site-directed mutagenesis, and two mutants were successfully obtained. The in vitro enzymatic reactions were carried out, and the differences in activity were detected by high performance liquid chromatography (HPLC). Finally, mutants T133LS134A and S339V with bifunctional activity were obtained. In addition to bifunctional activities of BAS and CHS, the modified PcPKS1 had much higher BAS activity than that of the wild type PcPKS1 under the conditions of pH 7.0 and pH 9.0, respectively. It provides a theoretical basis for future use of PcPKS1 in genetic engineering to regulate the biosynthesis of flavonoids and raspberry ketones.

Comparative analysis of differentially expressed genes for biosynthesis of active ingredients in fruits of different cultivars of Lycium barbarum L. based on transcriptome sequencing / 生物工程学报

To explore the differentially expressed genes (DEGs) related to biosynthesis of active ingredients in wolfberry fruits of different varieties of Lycium barbarum L. and reveal the molecular mechanism of the differences of active ingredients, we utilized Illumina NovaSeq 6000 high-throughput sequencing technology to conduct transcriptome sequencing on the fruits of 'Ningqi No.1' and 'Ningqi No.7' during the green fruit stage, color turning stage and maturity stage. Subsequently, we compared the profiles of related gene expression in the fruits of the two varieties at different development stages. The results showed that a total of 811 818 178 clean reads were obtained, resulting in 121.76 Gb of valid data. There were 2 827, 2 552 and 2 311 DEGs obtained during the green fruit stage, color turning stage and maturity stage of 'Ningqi No. 1' and 'Ningqi No. 7', respectively, among which 2 153, 2 050 and 1 825 genes were annotated in six databases, including gene ontology (GO), Kyoto encyclopedia of genes and genomes (KEGG) and clusters of orthologous groups of proteins (KOG). In GO database, 1 307, 865 and 624 DEGs of green fruit stage, color turning stage and maturity stage were found to be enriched in biological processes, cell components and molecular functions, respectively. In the KEGG database, the DEGs at three developmental stages were mainly concentrated in metabolic pathways, biosynthesis of secondary metabolites and plant-pathogen interaction. In KOG database, 1 775, 1 751 and 1 541 DEGs were annotated at three developmental stages, respectively. Searching the annotated genes against the PubMed database revealed 18, 26 and 24 DEGs related to the synthesis of active ingredients were mined at the green fruit stage, color turning stage and maturity stage, respectively. These genes are involved in carotenoid, flavonoid, terpenoid, alkaloid, vitamin metabolic pathways, etc. Seven DEGs were verified by RT-qPCR, which showed consistent results with transcriptome sequencing. This study provides preliminary evidences for the differences in the content of active ingredients in different Lycium barbarum L. varieties from the transcriptional level. These evidences may facilitate further exploring the key genes for active ingredients biosynthesis in Lycium barbarum L. and analyzing their expression regulation mechanism.

LC-MS analysis of 2-(2-phenylethyl) chromones in sodium chloride-treated suspension cells of Aquilaria sinensis / 中国中药杂志

Qualitative and quantitative analysis of 2-(2-phenylethyl) chromones in sodium chloride(NaCl)-treated suspension cells of Aquilaria sinensis was conducted by UPLC-Q-Exactive-MS and UPLC-QQQ-MS/MS. Both analyses were performed on a Waters T3 column(2.1 mm×50 mm, 1.8 μm) with 0.1% formic acid aqueous solution(A)-acetonitrile(B) as mobile phases at gradient elution. MS data were collected by electrospray ionization in positive ion mode. Forty-seven phenylethylchromones was identified from NaCl-treated suspension cell samples of A. sinensis using UPLC-Q-Exactive-MS, including 22 flindersia-type 2-(2-phenylethyl) chromones and their glycosides, 10 5,6,7,8-tetrahydro-2-(2-phenylethyl) chromones and 15 mono-epoxy or diepoxy-5,6,7,8-tetrahydro-2-(2-phenylethyl) chromones. Additionally, 25 phenylethylchromones were quantitated by UPLC-QQQ-MS/MS. Overall, the rapid and efficient qualitative and quantitative analysis of phenylethylchromones in NaCl-treated suspension cells of A. sinensis by two LC-MS techniques, provides an important reference for the yield of phenylethylchromones in Aquilariae Lignum Resinatum using in vitro culture and other biotechnologies.

Research progress on chemical constituents and pharmacological effects of Glechomae Herba and prediction of its Q-markers / 中国中药杂志

Glechomae Herba, the dried aerial part of Glechoma longituba(Labiatae), has the effects of promoting urination, draining dampness, and relieving stranguria. It has received wide attention in recent years owing to the satisfactory efficacy on lithiasis. Amid the in-depth chemical and pharmacological research, it has been found that Glechomae Herba has antibacterial, anti-inflammatory, antioxidant, antithrombotic, hepatoprotective, cholagogic, antitumor, hypoglycemic, and lipid-lowering effects. The main chemical constituents are volatile oils, flavonoids, terpenoids, phenylpropanoids, and organic acids. This paper summarized the chemical constituents and pharmacological effects of Glechomae Herba. Based on genetic relationship of plants, the characteristics, efficacy, and pharmacokinetics of the chemical constituents, and the potential of these constituents as quality markers(Q-markers), it was summed up that ursolic acid, caffeic acid, rosmarinic acid, luteolin-7-O-diglucuronide, apigenin, apigenin-7-O-diglucuronide, apigetrin, and glechone can be the candidate Q-markers of Glechomae Herba.

Phenylpropanoid pathway in plants and its role in response to heavy metal stress: a review / 生物工程学报

Phenylpropanoid metabolic pathway is one of the most important secondary metabolic pathways in plants. It directly or indirectly plays an antioxidant role in plant resistance to heavy metal stress, and can improve the absorption and stress tolerance of plants to heavy metal ions. In this paper, the core reactions and key enzymes of the phenylpropanoid metabolic pathway were summarized, and the biosynthetic processes of key metabolites such as lignin, flavonoids and proanthocyanidins and relevant mechanisms were analyzed. Based on this, the mechanisms of key products of phenylpropanoid metabolic pathway in response to heavy metal stress were discussed. The perspectives on the involvement of phenylpropanoid metabolism in plant defense against heavy metal stress provides a theoretical basis for improving the phytoremediation efficiency of heavy metal polluted environment.

Cloning and functional analysis of flavanone 3-hydroxylase gene in Rhododendron hybridum Hort / 生物工程学报

Flavanone 3-hydroxylase (F3H) is a key enzyme in the synthesis of phycocyanidins. In this experiment, the petals of red Rhododendron hybridum Hort. at different developmental stages were used as experimental materials. The R. hybridum flavanone 3-hydroxylase (RhF3H) gene was cloned using reverse transcription PCR (RT-PCR) and rapid-amplification of cDNA ends (RACE) techniques, and bioinformatics analyses were performed. Petal RhF3H gene expression at different developmental stages were analyzed by using quantitative real-time polymerase chain reaction (qRT-PCR). A pET-28a-RhF3H prokaryotic expression vector was constructed for the preparation and purification of RhF3H protein. A pCAMBIA1302-RhF3H overexpression vector was constructed for genetic transformation in Arabidopsis thaliana by Agrobacterium-mediated method. The results showed that the R. hybridum Hort. RhF3H gene is 1 245 bp long, with an open reading frame of 1 092 bp, encoding 363 amino acids. It contains a Fe2+ binding motif and a 2-ketoglutarate binding motif of the dioxygenase superfamily. Phylogenetic analysis showed that the R. hybridum RhF3H protein is most closely related to the Vaccinium corymbosum F3H protein. qRT-PCR analysis showed that the expression level of the red R. hybridum RhF3H gene tended to increase and then decrease in the petals at different developmental stages, with the highest expression at middle opening stage. The results of the prokaryotic expression showed that the size of the induced protein of the constructed prokaryotic expression vector pET-28a-RhF3H was about 40 kDa, which was similar to the theoretical value. Transgenic RhF3H Arabidopsis thaliana plants were successfully obtained, and PCR identification and β-glucuronidase (GUS) staining demonstrated that the RhF3H gene was integrated into the genome of A. thaliana plants. qRT-PCR, total flavonoid and anthocyanin contentanalysis showed that RhF3H was significantly higher expressed in the transgenic A. thaliana relative to that of the wild type, and its total flavonoid and anthocyanin content were significantly increased. This study provides a theoretical basis for investigating the function of RhF3H gene, as well as for studying the molecular mechanism of flower color in R. simsiib Planch.

Effect of processing method on chemical constituents of Rehmanniae Radix: based on UHPLC-LTQ-Orbitrap MS / 中国中药杂志

This study aims to explore the chemical composition of Rehmanniae Radix braised with mild fire and compare the effect of processing method on the chemical composition of Rehmanniae Radix. To be specific, ultra-high performance liquid chromatography with linear ion trap-orbitrap mass spectrometry(UHPLC-LTQ-Orbitrap MS) was used to screen the chemical constituents of Rehmanniae Radix. The chemical constituents were identified based on the relative molecular weight and fragment ions, literature information, and Human Metabolome Database(HMDB). The ion peak area ratio of each component before and after processing was used as the index for the variation. SIMCA was employed to establish principal component analysis(PCA) and orthogonal partial least squares discriminant analysis(OPLS-DA) models of different processed products. According to the PCA plot, OPLS-DA plot, and VIP value, the differential components before and after the processing were screened out. The changes of the content of differential components with the processing method were analyzed. A total of 66 chemical components were identified: 57 of raw Rehmanniae Radix, 55 of steamed Rehmanniae Radix, 55 of wine-stewed Rehmanniae Radix, 51 of repeatedly steamed and sundried Rehmanniae Radix Praeparata, 62 of traditional bran-braised Rehmanniae Radix, and 63 of electric pot-braised Rehmanniae Radix. Among them, the 9 flavonoids of braised Rehmanniae Radix were from Citri Reticulatae Pericarpium. PCA suggested significant differences in the chemical composition of Rehmanniae Radix Praeparata prepared with different processing methods. OPLS-DA screened out 32 chemical components with VIP value >1 as the main differential components. Among the differential components, 9 were unique to braised Rehmanniae Radix(traditional bran-braised, electric pot-braised) and the degradation rate of the rest in braised(traditional bran-braised, electric pot-braised) or repeatedly steamed and sundried Rehmanniae Radix was higher than that in the steamed or wine-stewed products. The results indicated the chemical species and component content of Rehmanniae Radix changed significantly after the processing. The 32 components, such as rehmapicrogenin, martynoside, jionoside D, aeginetic acid, hesperidin, and naringin, were the most important compounds to distinguish different processed products of Rehmanniae Radix. The flavonoids introduced by Citri Reticulatae Pericarpium as excipient may be the important material basis for the effectiveness of braised Rehmanniae Radix compared with other processed products.

Mechanism of total flavonoids of Rhododendra simsii in alleviating ischemic brain injury / 中国中药杂志

This study explores the effect of total flavonoids of Rhododendra simsii(TFR) on middle cerebral artery occlusion(MCAO)-induced cerebral injury in rats and oxygen-glucose deprivation/reoxygenation(OGD/R) injury in PC12 cells and the underlying mechanism. The MCAO method was used to induce focal ischemic cerebral injury in rats. Male SD rats were randomized into sham group, model group, and TFR group. After MCAO, TFR(60 mg·kg~(-1)) was administered for 3 days. The content of tumor necrosis factor-α(TNF-α), interleukin-1(IL-1), and interleukin-6(IL-6) in serum was detected by enzyme-linked immunosorbent assay(ELISA). The pathological changes of brain tissue and cerebral infarction were observed based on hematoxylin and eosin(HE) staining and 2,3,5-triphenyltetrazolium chloride(TTC) staining. RT-qPCR and Western blot were used to detect the mRNA and protein levels of calcium release-activated calcium channel modulator 1(ORAI1), stromal interaction molecule 1(STIM1), stromal intera-ction molecule 2(STIM2), protein kinase B(PKB), and cysteinyl aspartate specific proteinase 3(caspase-3) in brain tissues. The OGD/R method was employed to induce injury in PC12 cells. Cells were randomized into the normal group, model group, gene silencing group, TFR(30 μg·mL~(-1)) group, and TFR(30 μg·mL~(-1))+gene overexpression plasmid group. Intracellular Ca~(2+) concentration and apoptosis rate of PC12 cells were measured by laser scanning confocal microscopy and flow cytometry. The effect of STIM-ORAI-regulated store-operated calcium entry(SOCE) pathway on TFR was explored based on gene silencing and gene overexpression techniques. The results showed that TFR significantly alleviated the histopathological damage of brains in MCAO rats after 3 days of admini-stration, reduced the contents of TNF-α, IL-1, and IL-6 in the serum, down-regulated the expression of ORAI1, STIM1, STIM2, and caspase-3 genes, and up-regulated the expression of PKB gene in brain tissues of MCAO rats. TFR significantly decreased OGD/R induced Ca~(2+) overload and apoptosis in PC12 cells. However, it induced TFR-like effect by ORAI1, STIM1 and STIM2 genes silencing. However, overexpression of these genes significantly blocked the effect of TFR in reducing Ca~(2+) overload and apoptosis in PC12 cells. In summary, in the early stage of focal cerebral ischemia-reperfusion injury and OGD/R-induced injury in PC12 cells TFR attenuates ischemic brain injury by inhibiting the STIM-ORAI-regulated SOCE pathway and reducing Ca~(2+) overload and inflammatory factor expression, and apoptosis.

Anti-hyperuricemia activity and its mechanism of flavonoid extract from saffron floral bio-residues / 中国中药杂志

A hyperuricemic rat model induced by adenine and ethambutol was established to investigate the anti-hyperuricemia activity and its mechanism of the flavonoid extract from saffron floral bio-residues. Sixty-seven SD rats were randomly divided into control group, model group, positive control group, and flavonoid extract groups(with 3 doses), respectively, and each group contained 11 or 12 rats. The hyperuricemic model was established by continuous oral administration of adenine(100 mg·kg~(-1)) and ethambutol(250 mg·kg~(-1)) for 7 days. At the same time, the positive control group was given allopurinol(20 mg·kg~(-1) per day) and the flavonoid extract groups were given the flavonoid extract at doses of 340, 170 and 85 mg·kg~(-1) per day, respectively. On day 8, rat serum, liver, kidney, and intestinal tissues were collected, and the levels of uric acid in serum and tissue, the xanthine oxidase activities and antioxi-dant activities in serum and liver were evaluated, and the kidney histopathology was explored. In addition, an untargeted serum metabolomics study was performed. According to the results, the flavonoid extract effectively reduced the uric acid levels in serum, kidney and ileum and inhibited the xanthine oxidase activities and elevated the antioxidant activities of serum and liver in hyperuricemic rat. At the same time, it reduced the levels of inflammation factors in kidney and protected renal function. Moreover, 68 differential metabolites of hyperuricemic rats were screened and most of which were lipids and amino acids. The flavonoid extract significantly retrieved the levels of differential metabolites in hyperuricemic rats, such as SM(d18:1/20:0), PC[18:0/18:2(92,12Z)], palmitic acid and citrulline, possibly through the following three pathways, i.e., arginine biosynthesis, glycine, serine and threonine metabolism, and histidine metabolism. To sum up, the flavonoid extract of saffron floral bio-residues lowered the uric acid level, increased the antioxidant activity, and alleviated inflammatory symptoms of hyperuricemic rats, which may be related to its inhibition of xanthine oxidase activity and regulation of serum lipids and amino acids metabolism.