establishment of a rat model of skeletal muscle injury

Bir sıçan modelinde kas yaralanmasını tedavi etmek için Ding'in yuvarlanma manipülasyonuna dayalı bir masaj aleti

Muscle injuries are common traumatic injuries in clinical and daily life, caused by external blows (contusions) or chronic overstrain of muscle fibers (strains), etc., resulting in muscle dysfunction and pain, even seriously affecting the patient&#39;s quality of life1. Starting rehabilitation as early as possible after an acute strain injury is the key to reducing the time to return to sports2&#160;and in reducing pain3,4. In modern Western medicine, clinical first aid for muscle injuries follows the principles of rest, ice, compression, and elevation (RICE) to stop injurious bleeding into the muscle tissue5&#160;and non-steroidal anti-inflammatory drugs to relieve pain6. The discovery of novel therapies such as exosomes7&#160;and tissue engineering8&#160;became potential treatment strategies for skeletal muscle diseases, compensating for previous pharmacological treatments&#39; shortcomings. However, it can also increase the cost of treatment for patients, putting them under tremendous financial pressure9. Therefore, alternative and complementary therapies are recommended for treating musculoskeletal problems10. Tuina is widely used clinically in China as a traditional medical method and is popular among patients for its efficacy and fewer side effects. Tuina therapy for musculoskeletal disorders can alleviate pain and improve function11,12,13. Mr. Ding Jifeng, a famous Shanghai Tuina practitioner, founded Ding&#39;s roll method14. It is a unique rolling and crushing technique with a large force area, uniform and gentle force, and intense penetration.
Different animal models are based on different etiologies. They have advantages and disadvantages, and the selection of correct and appropriate animal models is of great significance to basic experiments, which helps understand the cellular and molecular signaling pathways of regeneration and repair after skeletal muscle injury to develop new therapies for treating the treatment of skeletal muscle diseases. Chemically induced models of muscle injury are widely used, with injections of skeletal muscle causing myofiber necrosis and producing regenerated areas that can effectively regenerate within 2 weeks15. Both notexin and bupivacaine can cause muscle damage. However, notexin can cause more severe myotoxic damage to skeletal muscle than bupivacaine, and natural functional recovery is relatively slower16. Drug intramuscular injection molding not only takes less time but also has controlled effects and extent of skeletal muscle damage. This quantifiable control makes successful molding less difficult15,17.
Inflammatory response is an essential biological response that has been extensively studied in the context of myopathy18,19. In the early stages of skeletal muscle injury, myofiber necrosis disrupts local muscle homeostasis, and many inflammatory cells infiltrate the injury site, secreting many pro-inflammatory cytokines19. Creatine kinase (CK) is a traditional serum biomarker for assessing skeletal muscle injury. However, it lacks tissue specificity20&#160;and sensitivity21, which limits its ability to assess the extent of drug-induced muscle damage and indirectly report the extent of muscle recovery after injury. Novel biomarkers, including fatty acid binding protein 3 (FABP3), have recently shown relatively high tissue specificity and sensitivity in rodent models of skeletal muscle injury. FABP3 is a family of binding proteins expressed primarily in cardiac and skeletal muscle cells and implicated in fatty acid metabolism, transport, and signaling22. Therefore, we chose a combination of two biomarkers, CK and FABP3, to assess the extent of notexin-induced skeletal muscle damage and recovery after treatment.
In rodents, the muscles are shallow, and the skin area is small, which also determines that the various parameters of massage in rodents will not be the same as in humans, such as in animal therapy, the massage therapist should treat them with less force using Ding&#39;s roll method, and may not be conducive to the operation of this technique due to the small size of the injured area, which can ultimately lead to a reduction in the effectiveness of the massage. Therefore, the experiment utilized the rolling massager made in-house, which conforms to the characteristics of Ding&#39;s roll method, to intervene and evaluate the therapeutic effect of the notexin-induced skeletal muscle injury model in rats, which helps to standardize the parameters of Tuina in experimental animal studies in order to profoundly investigate the molecular mechanism of action of Tuina, a traditional Chinese medicine treatment method, on musculoskeletal diseases.

Yazar Spotlight: Kendi Kendine Yapılan Bir Masaj Aleti Kullanarak Sıçanlarda Tuina Masajı Yapma 

Sıçanlarda Notexin Kaynaklı Kas Yaralanmasında Ding'in Rulo Yöntemini Taklit Etmek

Watch this Scientific Journal Video about Establishment of a Rat Model of Skeletal Muscle Injury at JoVE.com

Establishment of a Rat Model of Skeletal Muscle Injury  

İskelet kası yaralanmasının sıçan modelinin kurulması

Sıçanları, tuina gruplarıyla kontrol, notexin ve notexin dahil olmak üzere her biri sekiz sıçandan oluşan üç gruba rastgele bölerek başlayın. Fareyi uyuşturduktan sonra tüy dökücü krem ile sağ alt ekstremitedeki tüyleri alın. Daha sonra cildi dairesel bir dışa doğru hareketle üç alternatif iota, dört dezenfekte edici solüsyon ve% 75 alkol ile dezenfekte edin.İskelet kası yaralanma modelini oluşturmak için, notexin solüsyonunu intramüsküler olarak sadece bir bacakta gastroknemius kasına enjekte etmek için 30 gauge iğneli bir mililitrelik bir şırınga kullanın. İğneyi çıkarmadan önce üç saniye bekleyin. Kontrol grubundaki sıçanlara 200 mikrolitre salin solüsyonu enjekte edin.Anestezi uygulanmış fareleri temiz yataklı boş kafeslere taşıyın. Sıçanlar yeterli bilinci yeniden kazanana kadar doku rengini ve solunum hızını görsel olarak gözlemleyin. Fareleri ev kafesine geri koyun ve tipik olarak 24 saat boyunca yetiştirin.Yaralanma sonrası sıçan iskelet kasının morfolojik özellikleri hematoksilen ve eozin boyama ile gözlendi. Notexin enjeksiyonu ile yaralanan gastroknemius kası, kontrollere kıyasla birçok kas hücresinin yırtıldığını gösterdi. Hücreler atrofik, nekrotik ve düzensiz olarak düzenlenmiş, etkilenen bölgenin etrafına yüksek oranda nötrofil ve lenfosit infiltrasyonu yapmıştı.

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Research

JoVE Journal

Medicine

Establishment of a Rat Model of Skeletal Muscle Injury

153 Görüntüleme. Sıçanları, tuina gruplarıyla kontrol, notexin ve notexin dahil olmak üzere her biri sekiz sıçandan oluşan üç gruba rastgele bölerek başlayın. Fareyi uyuşturduktan sonra tüy dökücü krem ile sağ alt ekstremitedeki tüyleri alın. Daha sonra cildi dairesel bir dışa doğru hareketle üç alternatif iota, dört dezenfekte edici solüsyon ve% 75 alkol ile dezenfekte edin.İskelet kası yaralanma modelini oluşturmak için, notexin solüsyonunu intramüsküler olarak sade...

Video: Establishment of a Rat Model of Skeletal Muscle Injury  

Mimicking Ding's Roll Method on Notexin-Induced Muscle Injury in Rats

Ding's roll method is one of the most commonly used manipulations in traditional Chinese massage (Tuina) clinics and one of the most influential contemporary Tuina manipulations in China. It is based on the traditional rolling method commonly used in the one finger Zen genre and named Ding's roll method. Due to its anti-inflammatory and blood circulation-promoting effects, Ding's rolling method has sound therapeutic effects on myopathy. Because of the large area of force applied to human skin, Ding's roll method is challenging to perform on experimental animals with small skin areas, such as rats and rabbits. Additionally, the strength of Tuina applied to the human body differs from that applied to experimental animals, so it may happen that the strength is too high or too low to achieve the therapeutic effect of Tuina during the experiment. This experiment aims to create a simple massager suitable for rats based on Ding's rolling manipulation parameters (strength, frequency, Tuina duration). The device can standardize manipulation in animal experiments and reduce the variation in Tuina force applied to different animals due to subjective factors. A rat model of notexin-induced skeletal muscle injury was established, and plasma injury markers creatine kinase (CK) and fatty acid binding protein 3 (FABP3) were used to assess the therapeutic effect of Tuina on skeletal muscle injury. The results showed that this Tuina massager could reduce the levels of CK and FABP3 expression and slow down the degree of skeletal muscle injury. Therefore, the Tuina massager described here, mimicking Ding's roll method, contributes to standardizing Tuina manipulation in experimental research and is of great help for subsequent research on the molecular mechanism of Tuina for myopathy.

This protocol describes a simple device that mimics Ding's roll method, establishes a rat model of skeletal muscle injury, and uses hematoxylin-eosin staining to observe the pathology of damaged tissue and enzyme-linked immunosorbent assay to detect changes in serum damage markers.