Fixation and Operational Method for Abdominal Massage in T2DM Mice

Type two diabetes mellitus affects a large number of world population. My research explores how a new abdominal massage device affects blood glucose, lipid metabolism, and insulin sensitivity in T2DM mice. Recently, we developed a device for immobilized mice to perform massage, improve control, and the reducing stress for T2DM studies. We've found abdominal massage lower blood glucose, improves lipid metabolism, and boosts insulin sensitivity in T2DM mice, like metformin. Our protocol fills the gap in reproducible non-invasive methods to study abdominal massage effect on T2DM in mice. Our device offers precise control, less stress, and better consistence than manual or post assisted massage technique course. We will focus on fine-tuning massage pressure and the frequency plus testing the device on other metabolic disorders next.

[Narrator] Prepare a one millimeter thick polypropylene board to create a fixation plate. Shape the plate into a semicircle with a radius of 4.5 centimeters on one side and three centimeters on the other. When unfolded, the plate should form a trapezoidal shape with an upper base of eight centimeters, a lower base of 10 centimeters, and side lengths of 10 centimeters. Use a transparent plate to observe the mouse's fixation. Depending on the size of the mice, punch four long holes in the fixation plate. Then, obtain a soft and durable nonwoven fabric. Cut it into an elliptical shape at the center with four elongated straight strips at the corners, ensuring that these strips can pass through the four holes in the fixation plate. Based on the size of the mouse, cut four round holes in the fabric strips to allow the mouse's limbs to pass through. Next, select a pressure sensitive film sensor based on the mouse's abdominal size. Attach the pressure film to the section of the fixation fabric corresponding to the mouse's abdomen and connect the film to the pressure monitoring device. Pass the four long strips of the fixation fabric through the holes in the fixation plate, creating a passage through which the mouse can easily pass. Place the mouse on the side with the 4.5 centimeter radius. Use its burrowing instinct to encourage it to move toward the side with the three centimeter radius. Observe the mouse's position from the back of the transparent fixation plate. When the mouse reaches the appropriate position, tighten the four fabric strips to bind the mouse to the central part of the fixation fabric. Then, tie the strips in knots to secure the mouse. Now, flip the fixation plate over and use forceps to pull the mouse's limbs out of the four round holes in the fixation fabric, securing the mouse further. Hold the plate with one hand and position the fixed mouse device in the paw. Using the thumb, perform an abdominal massage on the mouse's abdomen. Closely monitor the pressure monitoring device to ensure the force applied to the pressure film sensor remains constant and uniform. After 20 minutes of abdominal massage, loosen the knots on the fabric strips, allowing the mouse to free itself from the fixation device. The fasting blood glucose and random blood glucose levels, as well as body weight, were significantly increased in the model group compared to the blank group. Both abdominal massage and metformin treatment reduced fasting blood glucose and random blood glucose levels in T2DM mice with no significant difference between the two treatments. Both interventions also improved body weight, indicating their potential role in blood glucose control. Abdominal massage significantly decreased total cholesterol, low density lipoprotein and triglyceride levels, and increased high density lipoprotein levels similar to metformin treatment. The expression levels of IRS-1, IRS-2, PI3K, AKT, and GLUT4 mRNA in liver tissues were significantly reduced in the model group compared to the blank group. Both abdominal massage and metformin significantly increased PI3K, AKT, and GLUT4 mRNA expression to almost the same extent.