Name: preparation of herbal medicine: er-xian decoction and er-xian-containing serum for in vivo and in vitro experiments
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Este trabajo fue apoyado por la Fundación Nacional de Ciencias Naturales de China (81573992). Damos las gracias a Emily K. Lo y Kathleen DiNapoli por su ayuda en la edición de idiomas.

El protocolo sigue las pautas del cuidado animal de la universidad de Shangai de la medicina china tradicional y es aprobado por el experimento animal Comité de Ética animal de Shangai. 1. Protocolo I: Preparación de EXD <ol><li> Cálculo <ol><li> Administrar el EXD a un grupo de tratamiento de 10 ratones hembra de seis meses de edad de Imprinting Control Region (ICR) (0,02 kg / ratón), 5 días a la semana durante 12 semanas. NOTA: Se necesitan un total de 1.702...

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S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+in+vitro+and+in+vivo+investigation+of+the+antimetastatic+effects+of+a+Chinese+medicinal+decoction,+erxian+decoction,+on+human+ovarian+cancer+models.">An in vitro and in vivo investigation of the antimetastatic effects of a Chinese medicinal decoction, erxian decoction, on human ovarian cancer models.</a> Integr Cancer Ther. 12 (4), 336-346 (2013).</li><li>Zhu, Z., Li, L., Jin, X., Fang, J., Zhang, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Er-Xian+Decoction,+a+traditional+Chinese+herbal+formula,+intervening+early+in+hypothalamic-pituitary+axis+of+male+rats+with+delayed+puberty.">Er-Xian Decoction, a traditional Chinese herbal formula, intervening early in hypothalamic-pituitary axis of male rats with delayed puberty.</a> Pharmacogn Mag. 10 (40), 517-521 (2014).</li><li>Miao, M. 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En los últimos años, se ha prestado más atención a las hierbas medicinales, un tipo de medicina alternativa que se ha aplicado en el entorno clínico en el mundo oriental durante miles de años. Diferente del patrón de &quot;banco a la cama&quot; de la medicina moderna, la medicina herbal tradicional primero requiere la &quot;cama al banco&quot; patrón para explicar sus mecanismos. Esto puede ser seguido por la validación realizada en la etapa de banca y el procesamiento para el desarrollo de fármacos recién op...

El interés en el estudio y la aplicación de la medicina herbaria tradicional está creciendo actualmente. A diferencia de las drogas modernas, en las cuales los ingredientes químicos son definidos, las fórmulas herbarias tienen algunos ingredientes desconocidos y requieren procesos de extracción para permitir la entrega de sus compuestos activos. Aunque muchos estudios tratan de seleccionar un pequeño compuesto con una estructura bien conocida como representante de toda la hierba o fórmula herbaria, ni las eficacias farmacológicas ni los mecanismos pueden considerarse equivalentes 1 , 2 . Mientras que las huellas dactilares permiten el análisis de los constituyentes de fórmulas herbarias complejas, algunos constituyentes todavía no se analizan claramente, lo que conduce a un desafío cuando se combinan todos los extractos para el estudio 3 . Las interacciones de numerosos constituyentes / extractos median los efectos terapéuticos de las hierbas medicinales. Para conservar esta ventaja, la forma tradicional de extracto-decocciónIon-todavía es ampliamente utilizado en la clínica. Dado que el procedimiento de extracción tiene un gran impacto en la eficacia terapéutica, es necesario un protocolo estándar de decocción tradicional de agua, especialmente para estudios in vivo 4 , 5 . Por otro lado, al explorar mecanismos farmacológicos, la administración de decocciones a base de hierbas durante estudios in vitro o ex vivo es también un desafío. El concepto de un suero que contenía fármacos fue propuesto por primera vez por Tashino en 1988 6 . Desde entonces, un número creciente de investigadores lo han aplicado a la medicina herbal 7 , 8 , 9 . Aunque el método del suero que contiene fármaco tiene algunas limitaciones, tales como la influencia de ciertos componentes del suero, todavía se considera que es un método que imita de cerca las condiciones fisiológicas. 10 , 11 , 12 , 13 . También se ha aplicado al tratamiento de la anemia aplásica 14 , osteoporosis menopáusica 15 , 16 , 17 , fallo ovárico prematuro 18 , cáncer de mama 19 , cáncer de ovario 20 y pubertad tardía 21 . Aquí presentamos protocolos detallados para la preparación de la decocción er-xian (EXD) y su suero que contiene fármaco. Además, se describe la aplicación de EXD y EXD conteniendo suero a los modelos de osteoporosis menopáusicas murinos. Un EXD se compone de 9 g cada uno de Curculigo orchioides Gaertn , Herbaa Epimedii </eM, Radix Morindae Officinalis y Radix Angelicae Sinensis y 6 g de Cortex Phellodendri y Rhizoma Anemarrhenae por paciente adulto al día. La dosis equivalente para un ratón es de 0,1418 EXD / kg / día basándose en la siguiente ecuación 22 : dB = dA * RB / RA * (WA / WB) 1/3 . DA y dB se refieren a la dosis por peso corporal (mg / kg) del ser humano y del ratón, respectivamente. La dosis, en mg / kg, se sustituye por el número de EXD / kg. RA y RB representan el factor del cuerpo humano y el factor corporal del ratón respectivamente, que son proporcionales a la (superficie corporal (m 2 ) / peso corporal (kg)) 2/3 (ver Tabla 1 ). WA y WB indican el peso corporal (kg) del humano y del ratón, respectivamente.

<table border="0" cellpadding="0" cellspacing="0" width="651">
	
	<tbody>
		<tr height="165">
			<td height="165" style="height:165px;width:216px;">Curculigo orchioides Gaertn (9&#8201;g), Herbaa Epimedii (9&#8201;g), Radix Morindae Officinalis (9&#8201;g), Radix Angelicae Sinensis (9&#8201;g), Cortex Phellodendri (6&#8201;g), and Rhizoma Anemarrhenae (6&#8201;g)</td>
			<td style="width:149px;">Kang-qiao Chinese Medicine Yinpian Co. Ltd (Shanghai, CN)</td>
			<td style="width:119px;">160922</td>
			<td style="width:167px;">EXD components</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:216px;">Filter paper</td>
			<td style="width:149px;">GElifesciences</td>
			<td style="width:119px;">99-103-952</td>
			<td style="width:167px;">Filter EXD decoction before concentration</td>
		</tr>
		<tr height="60">
			<td height="60" style="height:60px;width:216px;">Imprinting Control Region (ICR) mice</td>
			<td style="width:149px;">Shanghai Laboratory Animal Center</td>
			<td style="width:119px;">SCXK 2007-0005</td>
			<td style="width:167px;">In vivo study</td>
		</tr>
		<tr height="60">
			<td height="60" style="height:60px;width:216px;">Sprague Dawley rats</td>
			<td style="width:149px;">Shanghai Laboratory Animal Center</td>
			<td style="width:119px;">SCXK 2007-0005</td>
			<td style="width:167px;">EXD-containing serum preparation</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:216px;">Syringe filter</td>
			<td style="width:149px;">Millipore</td>
			<td style="width:119px;">SLGP033RB</td>
			<td style="width:167px;">0.22 &#181;m</td>
		</tr>
		<tr height="61">
			<td height="61" style="height:61px;width:216px;">gavage needles (10 ml)</td>
			<td style="width:149px;">Shanghai BO Ge trade sales department</td>
			<td align="right" style="width:119px;">59104274</td>
			<td style="width:167px;">Adminstration of EXD</td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;">Ketamine (80 mg/kg)&#160;</td>
			<td style="width:149px;">Fujian Gutian Pharma Co. Ltd</td>
			<td style="width:119px;">H35020148</td>
			<td>Anesthesia</td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:216px;">Xylazine (10 mg/kg)</td>
			<td style="width:149px;">Sunway Pharma Co. Ltd</td>
			<td style="width:119px;">CB07591</td>
			<td>Anesthesia</td>
		</tr>
		<tr height="81">
			<td height="81" style="height:81px;width:216px;">Dulbecco&#8217;s modification of Eagle&#8217;s medium Dulbecco (DMEM) culture medium</td>
			<td style="width:149px;">Gibco</td>
			<td style="width:119px;">12800-116</td>
			<td style="width:167px;">DMEM with 2 mM L-glutamine and without ribonucleosides and ribonucleotides</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Streptomycin</td>
			<td style="width:149px;">Sigma</td>
			<td align="right" style="width:119px;">1277</td>
			<td style="width:167px;">100 &#181;g / ml</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Penicillin</td>
			<td style="width:149px;">Sigma</td>
			<td align="right" style="width:119px;">4687</td>
			<td style="width:167px;">100 &#181;g / ml</td>
		</tr>
	</tbody>
</table>

preparation of herbal medicine: er-xian decoction and er-xian-containing serum for in vivo and in vitro experiments

La medicina herbal tradicional, una medicina alternativa en el ambiente clínico, ha recibido una atención creciente en los últimos años. Antes de la entrega al cuerpo, se requiere comúnmente un procedimiento de extracción adicional para liberar los componentes activos de las hierbas crudas. La decocción acuosa es un procedimiento de extracción clásico que todavía se utiliza ampliamente en los entornos clínicos. Aquí, proponemos un protocolo detallado para la decocción er-xian (EXD) con el fin de aplicar las decocciones a base de hierbas a los estudios experimentales. Se describe el cálculo de una dosis apropiada para animales, así como los cuatro pasos principales de EXD: remojo, decocción de agua, filtración y concentración. Además, el suero que contiene EXD se introduce a las ratas como un medio de validación in vitro . En este caso, las ratas fueron administradas por vía oral EXD durante tres días. Las muestras de sangre se recogieron entonces, se inactivaron, se centrifugaron y se filtraron. El suero, diluido con el medio de cultivo, puede utilizarse para tratar células o tejidos en vItro Por ejemplo, EXD se aplicó tanto a estudios in vivo e in vitro y demostró que EXD mejora la osteogénesis. Este protocolo puede utilizarse como referencia para la preparación y aplicación de medicamentos herbales.

El efecto de EXD sobre la densidad ósea de ratones OVX La tinción con hematoxilina y eosina (H &amp; E) de la sección de la vértebra lumbar muestra un aumento de las trabéculas óseas después del tratamiento EXD in vivo ( Figura 1A , panel derecho) en comparación con las del grupo OVX ( Figura 1A , panel izquierdo). La Figura 1B muestra las imágenes representativ...

Watch this Scientific Journal Video about Preparation of Herbal Medicine: Er-Xian Decoction and Er-Xian-containing Serum for In Vivo and In Vitro Experiments at JoVE.com

Preparation of Herbal Medicine: Er-Xian Decoction and Er-Xian-containing Serum for In Vivo and In Vitro Experiments

Aquí, se presenta la preparación de una decocción er-xian (EXD) en cuatro pasos-empapamiento, decocción, filtración y concentración- y demuestran la administración de un suero preparado para EXD a ratas. Estos métodos son aplicables al estudio in vivo e in vitro de las decocciones a base de hierbas como las medicinas tradicionales chinas.

Preparación de Herbal Medicine: Decocción de Er-Xian y suero que contiene Er-Xian para<em&gt; In Vivo</em&gt; Y<em&gt; In Vitro</em&gt; Experimentos

Traditional herbal medicine, an alternative medicine in the clinical setting, has received increased attention in recent years. Before delivery to the ...

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Preparation of Herbal Medicine: Er-Xian Decoction and Er-Xian-containing Serum for In Vivo and In Vitro Experiments

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