formulation of a gavage decoction using traditional chinese medicine and its administration into mice

Formulação de decocção de ervas e padronização de gavagem em camundongos com osteossarcoma

Decoction formula is the most commonly used dosage form of the traditional Chinese medicine and the liquid medicinal drugs1,2. The decoction formula of traditional Chinese medicine is processed by decocting the herbs in water after being soaked, followed by filtering to discard the dregs.
Because the decoction formula can be individualized based on clinical necessities, especially in line with the characteristics of evidence-based treatment in traditional Chinese medicine, this dosage form via trans-oral administration has been applied in clinical practice for thousands of years with specific and irreplaceable advantages3,4. The quality of the decoction formula is directly related to the clinical efficacy.&#160;Therefore, the decoction formula must be prepared based on the principles for the standardized decoction methods to ensure the active ingredients are maximally extracted and protected3. Meanwhile, decoction-ready medicines are the main form of raw Chinese herbs used in clinical practice; the quality standards and production methods of prescribed decoction-ready medicines must comply with the provisions of the Chinese Pharmacopoeia2. Moreover, the prescribed decoction-ready medicines must be soaked in cold drinking water for at least 30 min before being decocted in a special pharmaceutical pot by heating5. Studies have shown that most of the active ingredients can be extracted by decocting for two times, and the&#160;Code of Practice for the Administration of Decoction Rooms of Traditional Chinese Medicines in Medical Institutions also prescribes decoction to be done twice4,6,7. If special Chinese herbs, such as one with hard texture or tonics, are prescribed, the decoction time is recommended to be extended. The decoction utensils and storage containers that directly come in contact with the decoction formula, starting from the container in which the prescribed decoction-ready medicines are soaked, should be chemically stable and have lids, such as casseroles, ceramics, glass and stainless steel. However, aluminum and ordinary plastic products should not be used, while iron and other corrosive utensils should be prohibited to avoid possible chemical reactions, which will reduce therapeutic efficacy and even produce harmful effects4,6,7.
Due to the limitations of performing clinical experiments, small animals bearing the human diseases, such as mice, are often used in medical research to explore the therapeutic efficacy and comprehensive mechanisms of different interventions including the decoction formula of traditional Chinese medicine8,9; however, unlike humans, the decoction formulas should be prepared every day, mixed together, and freshly administered 2x a day in clinical practice. Animals are unable to voluntarily take their medications on time with the required dosage, and most of the animal experiments related to traditional Chinese medicine are gavage-based oral administration of the decoction formulas at present. The decoction formulas used for in vivo animal experiments should be prepared in advance, aliquoted, stored at -20 &#176;C and thawed just before use3,4,10. Moreover, substandard animal immobilization and gavage needle manipulation increase the risk of unnecessary injuries and negative emotions. Therefore, as an important method of trans-oral drug delivery, the skilled gavage technique is particularly important to avoid possible esophageal injury and drug spillage, thus ensuring that each model animal receives an equal amount of prescribed herbal medicine to achieve accurate experimental results. Here, we provide a gavage protocol for the decoction formula of traditional Chinese medicine.

Autor em destaque: Protocolo padronizado de decocção de ervas para estudos aprimorados em animais

Estratégia de Gavagem para Fórmula de Decocção da Medicina Tradicional Chinesa em Camundongos Modelo de Osteossarcoma

This study provides a detailed and standardized protocol for about precaution preparation under gradual operation that's providing a reference for those in need. Our research ensures that each animal is dosed equally to produce accurate results. Our study aims to maximize the animal protection and improve the reliability of results from studies of about Decoction formulations.

Formulação de uma decocção de gavagem usando medicina tradicional chinesa e sua administração em camundongos

Para começar, mergulhe 52,5 gramas de hedyotis diffusa e 31,5 gramas da erva da calota craniana farpada em água por 30 minutos. Aqueça as ervas para ferver em fogo forte e, em seguida, aqueça por 15 a 25 minutos em fogo brando. Em seguida, use elásticos para fixar três camadas de gaze sobre a boca de um copo medidor.Filtre a decocção através da gaze. Despeje a decocção filtrada em um recipiente. Da mesma forma, faça uma segunda decocção fervendo as ervas em água potável apenas o suficiente para submergir as ervas.Depois de ferver, despeje a segunda decocção no mesmo recipiente que a primeira. Agora mergulhe 42 gramas de ginseng em água potável cerca de três centímetros sobre a superfície das ervas. Ferva o ginseng embebido em fogo forte por 60 minutos.Em seguida, despeje esta primeira decocção no recipiente com as outras decocções. Faça uma segunda decocção de ginseng na água potável apenas o suficiente para submergir a erva. Aqueça esta decocção em fogo brando por 40 minutos e filtre-a no recipiente de decocção.Ferva a decocção filtrada em fogo forte em uma panela. Em seguida, abaixe a chama para um fogo leve. Continue fervendo a decocção até obter a concentração desejada.Pipetar 21 mililitros da decocção concentrada em tubos de acordo com a dosagem necessária por uma semana. Guarde as alíquotas em geladeira a menos 20 graus Celsius. Coloque um mouse na superfície de uma gaiola de mouse.Agarre a cauda com o polegar e o indicador da mão esquerda. Em seguida, segure o pescoço do mouse com os dedos polegar, indicador e médio da mão direita. Segure o mouse para alinhar a cabeça, o pescoço e o corpo em linha reta.Insira a agulha de gavagem carregada na boca do rato a partir do canto e empurre a língua contra a mandíbula superior. Mantenha o corpo do rato no mesmo alinhamento e empurre suavemente a agulha de gavagem em direção ao estômago ao longo do esôfago. Injete a fórmula de decocção no estômago, remova a agulha e solte o mouse de volta na gaiola.A administração da decocção medicinal reduziu o volume tumoral do osteossarcoma em camundongos.

Research

JoVE Journal

Medicine

Formulation of a Gavage Decoction Using Traditional Chinese Medicine and its Administration into Mice

Gavage Strategy for Decoction Formula of Traditional Chinese Medicine in Osteosarcoma Model Mice

Decoction formula is the most commonly used dosage form in traditional Chinese medicine and applied in clinical practice for thousands of years by ...

Medicina