establishment of murine chronic root ganglion compression model

פרוטוקול Tuina סטנדרטי עבור מודל דחיסת DRG של חולדות

In clinical settings, it is common to observe neurological pathological pain caused by nerve root compression due to various reasons. The most typical form of this neuropathic pain is lumbar disc herniation (LDH), which is often persistent, recurrent, and difficult to cure. Approximately 9% of the global population is affected by LDH, leading to significant social and economic burdens1. The incidence of this type of neuropathic pain is increasing yearly, with a trend toward younger patients, due to changes in human production and lifestyle2. Despite the use of non-steroidal painkillers, patients&#39; symptoms cannot be completely alleviated. As a result, alternative therapies, such as Tuina, for treating pain caused by LDH have gained increasing attention.
Tuina therapy, a form of conservative treatment for LDH, is widely recommended in various clinical practice guidelines worldwide for preventing and treating lower back pain3,4. Research has shown that Tuina can significantly lower inflammatory factors such as serum IL-6 and tumor necrosis factor-alpha (TNF-&#945;) levels in LDH patients while improving patients&#39; pain and lumbar function impairment5. However, the specific mechanism behind Tuina therapy&#39;s pain-relieving effects remains unclear.
Animal models are a valuable tool for studying neuropathic pain caused by LDH6. They allow for behavioral measurements to evaluate the effectiveness of Tuina therapy and provide samples of the pathological physiology of LDH. For example, samples from the dorsal root ganglia in the thigh can be taken to verify changes in dorsal root ganglion cells. The chronic compression of the Dorsal Root Ganglion (CCD) model is commonly used to evaluate the pathological physiology of LDH, as it causes damage to the morphology of dorsal root ganglion cells that are consistent with the pathological changes seen in clinical cases of nerve compression caused by disc herniation7.
Many scholars have conducted several animal experiments on acupressure analgesia8,9,10. However, when implementing acupressure operations on animal models, they often imitate human acupressure. The therapeutic effect of acupressure is affected by factors such as the size, frequency, and the direction of the applied force11,12,13. If the experiment lacks a unified acupressure standard, such as the force, frequency, and duration of the operation, this may cause some deviation in the experimental results. This article introduces a set of acupressure treatment plans based on the characteristics of CCD rats, and promotes the development of standardized acupressure operations in animal models.

מחבר זרקור: אפקט משכך כאבים של טווינה על מודלים של חולדות עם דחיסה של כאב גנגליון שורש הגב  

אפקט משכך כאבים של Tuina על מודלים חולדה עם דחיסה של כאב גנגליון שורש הגב

Tuina analgesia is widely used in clinical practice. Our research focuses on the mechanism of action of Tuina analgesia, and investigate how Tuina achieve its analgesic effect. Achieving standardization in Tuina is currently the most significant obstacle.The effectiveness of Tuina is influenced by several factors such as treatment, area type, intensity, and frequency. The current protocol is optimized in two aspect based on the earlier research. Firstly, the force and frequency of pushing and holding operations were standardized and controlled by force-measuring gloves.Secondly, the results of imaging and animal behavior test were integrated to assess the results of modeling.

Watch this Scientific Journal Video about Establishment of Murine Chronic Root Ganglion Compression Model at JoVE.com

Establishment of Murine Chronic Root Ganglion Compression Model  

הקמת מודל דחיסת גנגליון שורש כרוני מורין

כדי לבסס את מודל החיה, התחל על ידי אבטחת חולדה מורדמת על לוח קצף, הידוק הגפיים והחותכות שלה באמצעות גומיות. נגבו את האזור המוכן במקלון אלכוהול בשיעור 75%. לאחר מכן, השתמשו במספריים כדי לבצע חתך של שניים עד שלושה סנטימטרים דרך העור, הפאשיה השטחית והפאשיה העמוקה שכבה אחר שכבה.אתר את עמוד השדרה האיליאק העליון הקדמי, ואחריו את התהליכים עמוד השדרה השלישי והרביעי. הדקו את התהליך הספינוסי עם מלקחיים עם שיניים והרימו אותו כדי לאפשר למספריים להיות קרובים לצד הימני של תהליך עמוד השדרה. חתכו את השריר המחובר לצד ימין של תהליך עמוד השדרה.לאחר מכן נתחו בבוטות את השריר המחובר למשטח החיצוני של לוחית החוליות עד שתורגש התנגדות. באופן דומה, לנתח בבוטות את השריר ואת fascia על המפרק zygapophysial. השתמש בבדיקה בצורת L כדי לאתר את הפתח הבין חולייתי.לאחר מכן, הכנס מוט נירוסטה בצורת L לתוך הפתח הבין חולייתי המותני הרביעי והחמישי. המשיכו לתפור את השריר, הפאשיה והעור שכבה אחר שכבה באמצעות 3-0 תפרים. הניחו את החולדה בקופסה תרמוסטטית עד שהיא מתעוררת והתבוננו בתפקוד הגפה האחורית הימנית של החולדה כדי להבטיח נורמליות.

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Research

JoVE Journal

Medicine

Establishment of Murine Chronic Root Ganglion Compression Model

144 Views.  Yueyang Hospital of Integrated Traditional Chinese and Western Medicine Affiliated to Shanghai University of Traditional  Chinese Medicine. כדי לבסס את מודל החיה, התחל על ידי אבטחת חולדה מורדמת על לוח קצף, הידוק הגפיים והחותכות שלה באמצעות גומיות. נגבו את האזור המוכן במקלון אלכוהול בשיעור 75%. לאחר מכן, השתמשו במספריים כדי לבצע חתך של שניים עד שלושה סנטימטרים דרך העור, הפאשיה השטחית והפאשיה העמוקה שכבה אחר שכבה.

Video: Establishment of Murine Chronic Root Ganglion Compression Model  

Analgesic Effect of Tuina on Rat Models with Compression of the Dorsal Root Ganglion Pain

Neuropathic pain is a prevalent condition that affects 6.9%-10% of the population and results from nerve damage due to various etiologies, such as lumbar disc herniation, spinal canal stenosis, and intervertebral foramen stenosis. Although Tuina, a traditional Chinese manual therapy, has shown analgesic effects in clinical practice for the treatment of neuropathic pain, its underlying neurobiological mechanisms remain unclear. Animal models are essential for elucidating the basic principles of Tuina. In this study, we propose a standardized Tuina protocol for rats with compression of the dorsal root ganglion (DRG), which involves inducing DRG compression by inserting a stainless steel rod into the intervertebral foramen, performing Tuina manipulation with specific parameters of location, intensity, and frequency in a controlled environment, and assessing the behavioral and histopathological outcomes of Tuina treatment. This article also discusses the potential clinical implications and limitations of the study and suggests directions for future research on Tuina.

This article presents a manipulation for treating chronic compression of the dorsal root ganglion in rats using Tuina therapy, along with a method for evaluating its effectiveness based on pain behavior and histopathological results.

Neuropathic pain is a prevalent condition that affects 6.9%-10% of the population and results from nerve damage due to various etiologies, such as lumbar ...

To establish the animal model, start by securing an anesthetized rat on a foam board, fastening its limbs and incisors using rubber bands. Wipe the prepared area with a 75%alcohol swab. Next, use scissors to make a two to three centimeter incision through the skin, superficial fascia, and deep fascia layer by layer.Locate the anterior superior iliac spine, followed by the third and fourth spinous processes. Clamp the spinous process with toothed forceps and lift it to allow the scissors to be close to the right side of the spinous process. Cut the muscle attached to the right side of the spinous process.Then bluntly dissect the muscle attached to the outer surface of the vertebral plate until resistance is felt. Similarly, bluntly dissect the muscle and fascia on the zygapophysial joint. Use an L-shaped probe to locate the intervertebral foramen.Next, insert an L-shaped stainless steel rod into the fourth and fifth lumbar intervertebral foramen. Proceed to suture the muscle, fascia, and skin layer by layer using 3-0 sutures. Place the rat in a thermostatic box until it wakes up and observe the function of the rat's right hind limb to ensure normalcy.

Tuina Therapy Administration to Murine Root Ganglion Compression Model

To standardize the tuina therapy, first, practice tuina using the finger sleeves feedback data, adjusting force to 5 newtons and frequency to 2 hertz. Next, perform tuina on immobilized rats, maintaining consistent force and frequency. Identify the acupoint by selecting the gastrocnemius muscle of the right hind limb at the junction of its two heads, approximately at BL 57.Perform the tuina by positioning the thumb vertically on the BL 57 acupoint and moving in rhythmic, small-range rotary motions exerting 5 newtons of pressure. After 17 days of tuina therapy, a significant difference was observed in paw withdrawal thresholds between the CCD rats receiving tuina therapy and the untreated CCD group. The rats in the CCD group receiving tuina therapy showed improvement in pain threshold from the beginning of treatment.Thermal pain thresholds differed significantly between the CCD group and the tuina group from day 14.

מתן טיפול Tuina למודל דחיסת גנגליון שורש מורין

Murine Pain Behavioral Testing to Study the Effect of Tuina Therapy

To conduct the Von Fray test, begin by placing the rats in a transparent tempered glass compartment on a metal wire grid stand at a height of 40 centimeters. Next, measure the mechanical withdrawal threshold with Von Frey electronic fibers. Stimulate the rat's foot until it reacts noticeably, such as by lifting the leg or avoiding it.To assess the paw withdrawal latency, or PWL using the Hargreaves method, focus the spotlight on the center of the rat's foot and press the start button. Monitor the rat for behaviors such as foot retraction or paw licking, and press the start button again to record the time. Perform the test on the same hind limb three consecutive times.After 17 days of Tuina therapy, a significant difference was observed in paw withdrawal thresholds between the CCD rats receiving Tuina therapy and the untreated CCD group. The rats in the CCD group receiving Tuina therapy showed improvement in pain threshold from the beginning of treatment. Thermal pain thresholds differed significantly between the CCD group and the Tuina group from day 14.